Tiam1、DcR3、Kallikrein6在大肠癌中的表达及与外周血微转移的关系
|
摘要
第一部分大肠癌患者外周血微转移检测的临床意义
目的大肠癌是最常见的恶性肿瘤之一,血行转移是大肠癌根治性手术失败的主要原因之一,也是患者死亡的主要原因。在接受了根治术的患者中,仍有超过50%的患者最终死于肿瘤的复发和转移,提示手术时可能已有大肠癌微转移的存在。本研究以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,探讨了大肠癌患者外周血微转移检测的临床意义,手术操作和外周血微转移的关系,以及大肠癌组织中微血管密度(microvessel density,MVD)和微淋巴管密度(lymphatic microvessel density,LMVD)与大肠癌外周血微转移的关系。
方法以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,采用RT-PCR技术检测大肠癌患者术前和术中外周血微转移;采用免疫组织化学方法检测大肠癌组织中MVD和LMVD;分析大肠癌患者外周血微转移、大肠癌组织中MVD和LMVD与临床病理参数的关系以及大肠癌组织中MVD和LMVD与外周血微转移的关系。
结果51例大肠癌患者中18例手术前外周血CK 20mRNA呈阳性表达,总阳性表达率为35.29%(18/51),在Ⅰ～Ⅳ期患者中的阳性表达率分别为12.50%(1/8)、23.53%(4/17)、35.29%(6/17)和77.78%(7/9),CK20 mRNA在外周血中的阳性表达与TNM分期相关(P＜0.05);手术中有21例呈阳性表达(21/51),3例术前阴性表达者术中转为阳性表达,均为直肠癌手术患者;CK20 mRNA在外周血中的阳性表达与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值、肿瘤分化程度、浸润深度无关(P＞0.05),但与淋巴结转移、远处转移、TNM分期相关(P＜0.05);MVD与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值无关(P＞0.05),但与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关(P＜0.05);LMVD与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值、肿瘤分化程度、浸润深度无关(P＞0.05),但与淋巴结转移、远处转移、TNM分期相关(P＜0.01);大肠癌患者外周血CK20 mRNA阳性表达组MVD和LMVD的值分别为58.83±18.28和9.72±1.64,而外周血CK20mRNA阴性表达组MVD和LMVD的值分别为30.48±9.31和8.21±1.87,显著低于阳性表达组(P＜0.01)。
结论大肠癌细胞外周血微转移是大肠癌的早期事件,外周血微转移发生率随临床病理分期的增加而升高,外周血微转移与淋巴结转移、远处转移、TNM分期相关,是反映大肠癌生物学行为的指标之一;手术操作有可能促进直肠癌患者外周血微转移的发生;MVD与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关;LMVD与淋巴结转移、远处转移、TNM分期相关;大肠癌组织血管化程度和淋巴管生成与外周血微转移密切相关,通过对二者的检测有助于判断外周血微转移发生的情况。
第二部分Tiam1、DcR3在大肠癌中表达的临床意义及与大肠癌外周血微转移的关系
目的侵袭和转移是恶性肿瘤的重要特征,也是肿瘤患者死亡的主要原因,因此对肿瘤的侵袭和转移及其相关基因的研究具有重要意义。T淋巴瘤侵袭转移诱导因子1(T lymphoma invasion and metastasis factor 1,Tiam1)与肿瘤侵袭和转移密切相关。DcR3(decoy receptor 3)是肿瘤坏死因子受体超家族成员,在多种人类恶性肿瘤中出现过度表达,且DcR3的表达状态与恶性肿瘤的淋巴结转移和病理分期密切相关。关于Tiam1和DcR3与大肠癌外周血微转移的研究报道不多,我们通过检测Tiam1和DcR 3在大肠癌组织和正常大肠粘膜组织中的表达,探讨了Tiam1和DcR3表达与大肠癌临床病理参数的关系以及二者与大肠癌外周血微转移的关系。
方法以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,采用RT-PCR方法检测大肠癌患者外周血微转移;采用实时荧光定量RT-PCR方法检测大肠癌组织和正常组织中Tiam1和DcR3的表达;分析大肠癌组织中Tiam1和DcR3表达与临床病理参数以及患者外周血微转移的关系。
结果51例大肠癌组织及正常粘膜组织中均见Tiam1 mRNA表达,表达水平分别为(20.61±4.99)×10~(-4)和(3.49±2.74)×10~(-4),大肠癌组织中Tiam1mRNA表达明显高于正常组织(P＜0.01);Tiam1 mRNA在伴有外周血微转移的大肠癌组织中的表达水平为(72.11±2.38)×10~(-4),在不伴外周血微转移的大肠癌组织中的表达水平为(15.78±5.01)×10~(-4),二者差异有统计学意义(P＜0.05);大肠癌组织中Tiam1 mRNA表达水平与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值无关(P＞0.05),但与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关(P＜0.05);51例大肠癌组织及正常粘膜组织中均见DcR3 mRNA的表达,表达水平分别为(42.40±2.83)×10~(-4)和(12.59±2.55)×10~(-4),大肠癌组织中DcR3 mRNA表达明显高于正常组织(P＜0.01);DcR3 mRNA在伴有外周血微转移的大肠癌组织中表达水平为(61.53±2.60)×10~(-4),在不伴外周血微转移的大肠癌组织中表达水平为(34.60±2.84)×10~(-4),但二者差异无统计学意义(P＞0.05);DcR3 mRNA的表达水平与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值无关(P＞0.05),但与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关(P＜0.05)。
结论大肠癌组织的Tiam1 mRNA和DcR3 mRNA高表达与大肠癌的发生相关;大肠癌组织的Tiam1 mRNA和DcR3 mRNA表达水平与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关;Tiam1 mRNA高表达是大肠癌患者发生外周血微转移的高危因素;但DcR3 mRNA高表达不能作为大肠癌患者发生外周血微转移的高危因素。
第三部分Kallikrein6在大肠癌中表达的临床意义及与大肠癌外周血微转移的关系
目的激肽释放酶(Kallikreins,KLK)是一组分泌型丝氨酸蛋白酶,存在于多种组织和生物体液中。人KLK基因家族参与恶性肿瘤的发生发展,多种KLK与前列腺癌、卵巢癌、乳腺癌和睾丸癌等内分泌相关肿瘤的发生发展密切相关,是此类肿瘤的潜在标志物。KLK6异常表达与多种肿瘤相关。国内目前未见有关KLK6和大肠癌关系的报道。我们通过检测KLK6 mRNA在大肠癌组织和正常大肠粘膜组织中的表达,探讨了KLK6表达与大肠癌临床病理资料的关系及其与外周血微转移的关系。
方法以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,采用RT-PCR方法检测大肠癌患者外周血微转移;采用实时荧光定量RT-PCR方法检测大肠癌组织和正常组织中KLK6的表达;分析大肠癌组织中KLK6表达与临床病理参数以及患者外周血微转移的关系。
结果51例大肠癌组织及正常粘膜组织中均见KLK6 mRNA的表达。大肠癌组织中和正常粘膜组织中KLK6 mRNA表达水平分别为(33.11±2.34)×10~(-4)和(12.88±2.51)×10~(-4),有高度统计学差异(P＜0.01);伴有外周血微转移的大肠癌组织中和不伴外周血微转移的大肠癌组织中KLK6 mRNA的表达水平分别为(40.64±2.05)×10~(-4)和(30.06±2.45)×10~(-4),差异无统计学意义(P＞0.05);大肠癌组织的KLK6 mRNA表达与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值、肿瘤分化程度、浸润深度、远处转移无关(P＞0.05),但与淋巴结转移、TNM分期相关(P＜0.05)。
结论大肠癌组织的KLK6 mRNA高表达与大肠癌的发生相关;大肠癌组织的KLK6 mRNA表达水平与淋巴结转移、TNM分期相关,与外周血微转移不相关,大肠癌组织的KLK6 mRNA的高表达不能作为大肠癌患者发生外周血微转移的高危因素。
Part One Clinical significance of peripheral blood micro-metastasis in colorectal cancer
Objective Peripheral blood micro-metastasis is one of the important factors for failure of surgical resection in colorectal cancer.Above 50%of the patients underwent surgical resection died of cancerous relapse or metastasis,indicating that micro-metastasis may have exited before the sugeries.CK20 mRNA has thought to be a specific biomarker for early diagnosis of micro-metastasis in colorectal cancer.To explore the role of determination of peripheral blood micro-metastasis in prognosis of colorectal cancer and to explore whether microvessel density(MVD) and lymphatic microvessel density(LMVD) have correlation with peripheral blood micro-metastasis.
Methods Using CK20 mRNA as a specific biomarker of tumor cells in peripheral blood,we determined its levels before and after surgical resection with the RT-PCR technique,also we determined the MVD and LMVD levels by immunohistochemistry.
Results Eighteen of the 51 patients were found positive CK20 mRNA expressions before surgical resection,of them,1(12.50%) was of the 8 patients with stageⅠ,4 (23.53%) was of the 17 patients with stageⅡ,6(35.29%) was of the 17 patients with stageⅢ,and 7(77.78%)was of the 9 patients with stageⅣ.These findings indicated that CK20 mRNA expression in peripheral blood correlated with the TNM classification in colorectal cancer.After surgical resection,3 more patients with rectum cancer were also shown positive CK20 mRNA expressions.CK20 mRNA expression had no relations to age,gender,primary location,tumor size,peripheral blood carcino-embryonic antigen level,and tumor differentiation and invasion,but had relations to lymphonode metastasis,distant metastasis and tumor TNM stage (P＜0.05).The MVD levels had no relations to age,gender,primary location,tumor size and the peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphonode metastasis,distant metastasis,tumor TNM stage(P＜0.05).Also the LMVD levels had no relations to age,gender,primary location,tumor size,the peripheral blood carcino-embryonic antigen level,and tumor differentiation and invasion,but had relations to lymphonode metastasis,distant metastasis,and tumor TNM stage(P＜0.05).The MVD and LMVD levels were 58.83±18.28 and 9.72±1.64 in the positive CK20 mRNA group,however they were 30.48±9.31 and 8.21±1.87 in the negative CK20 mRNA group,the difference was statistically significant(P＜0.01 ).
Conclusions Peripheral blood micro-metastasis is related to lymphnode metastasis, distant metastatis and TNM stage.Surgical procedures may promote the genesis of periphral blood micro-metastasis.Determination of tissue vascularization and genesis of lymphatic vessel is helpful in judging genesis of peripheral blood micro-metastasis in colorectal cancer.
Part Two Tiam1 and DcR3 expressions in colorectal cancer and their correlation with peripheral blood micro-metastasis
Objective Invasion and metastasis are key characteristics of malignant tumors.It has been shown that T lymphoma invasion and metastasis factor 1(Tiam1) closely correlates with tumor invasion and metastasis.Decoy receptor 3(DcR3) is one member of the tumor necrosin receptor hyperfamily,and it is overexpressed in many human malignant tumors,also its overexpression closely relates to tumor lymphnode metastasis and pathologic staging.To determine the expressions of Tiam1 mRNA and DcR3 mRNA in cancerous tissues and non-cancerous tissues of colorectal cancer,and to explore their correlation with pathologic data and peripheral blood micro-metastasis.
Methods Using the RT-PCR method,the CK20 mRNA expression was determined.Using fluorescence quantitative-RT-PCR(FQ-RT-PCR),the expressions of Tiam1 mRNA and DcR3 mRNA were determined in 51 cases of colorectal carcinoma.
Results The Tiam1 mRNA expression level was(20.61±4.99)×10~(-4) in cancerous tissues and was(3.49±2.74)×10~(-4) in non-cancerous tissues(P＜0.01).Also it was (72.11±2.38)×10~(-4) in cancerous tissues with micro-metastasis and was(15.78±5.01)×10~(-4) in cancerous tissues without micro-metastasis,the difference was statistically significant(P＜0.05).Tiam1 mRNA expression had no relations to age,gender, primary location,tumor size and the peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphnode metastasis, distant metastasis and tumor TNM stage(P＜0.05).The DcR3 mRNA expression level was(42.40±2.83)×10~(-4) in cancerous tissues and was(12.59±2.55)×10~(-4) in non-cancerous tissues(P＜0.01).Also it was(61.53±2.60)×10~(-4) in cancerous tissues with micro-metastasis and was(34.60±2.84)×10~(-4) in cancerous tissues without micro-metastasis,but the difference was not statistically significant(P＞0.05).DcR3 mRNA expression had no relations to age,gender,primary location,tumor size and peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphnode metastasis,distant metastasis and tumor TNM stage(P＜0.05).
Conclusions Overexpressions of Tiam1 mRNA and DcR3 mRNA correlate with genesis of colorectal cancer.Tiam1 mRNA overexpression is a high risk factor for peripheral blood micro-metastasis in colorectal cancer,but DcR3 mRNA can not be used as a high risk factor for peripheral blood micro-metastasis in colorectal cancer.
Part Three Human KaUikrein6 expression in colorectal cancer and its correlation with peripheral blood micro-metastasis
Objective Kallikreins(KLKs) have a similar genomic organization and show significant homology at both the nucleotide and the protein level.Human KLK gene family has participated in genesis and development of cancers.A growing body of evidence suggests that KLKs are involved in human malignancies and that many KLKs are promising biomarkers of prostate,ovarian,breast and testicular cancers.To determine the expression of KLK6 mRNA in colorectal cancer and to explore its relationship to clinicopathologic features and peripheral blood micro-metastasis in colorectal cancer.
Methods Using the RT-PCR method,the CK20 mRNA expression was determined.Using the FQ-RT-PCR method,the KLK6 mRNA expression was determined in 51 cases of colorectal carcinoma.
Results The KLK6 mRNA expression level was(33.11±2.34)×10~(-4) in cancerous tissues and was(12.88±2.51)×10~(-4) in non-cancerous tissues(P＜0.01).Also it was (40.64±2.05)×10~(-4) in cancerous tissues with micro-metastasis and was(30.06±2.45)×10~(-4) in cancerous tissues without micro-metastasis,the difference was not statistically significant(P＞0.05).KLK6 mRNA expression had no relations to age, gender,primary location,tumor size,peripheral blood carcino-embryonic antigen level,tumor differentiation and invasion,and distant metastasis,but had relations to lymphnode metastasis and tumor TNM stage(P＜0.05).
Conclusions Overexpression of KLK6 mRNA correlates with genesis of colorectal cancer,however,it can not be used as a high risk factor of peripheral blood micro-metastasis in colorectal cancer.
目的大肠癌是最常见的恶性肿瘤之一,血行转移是大肠癌根治性手术失败的主要原因之一,也是患者死亡的主要原因。在接受了根治术的患者中,仍有超过50%的患者最终死于肿瘤的复发和转移,提示手术时可能已有大肠癌微转移的存在。本研究以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,探讨了大肠癌患者外周血微转移检测的临床意义,手术操作和外周血微转移的关系,以及大肠癌组织中微血管密度(microvessel density,MVD)和微淋巴管密度(lymphatic microvessel density,LMVD)与大肠癌外周血微转移的关系。
方法以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,采用RT-PCR技术检测大肠癌患者术前和术中外周血微转移;采用免疫组织化学方法检测大肠癌组织中MVD和LMVD;分析大肠癌患者外周血微转移、大肠癌组织中MVD和LMVD与临床病理参数的关系以及大肠癌组织中MVD和LMVD与外周血微转移的关系。
结果51例大肠癌患者中18例手术前外周血CK 20mRNA呈阳性表达,总阳性表达率为35.29%(18/51),在Ⅰ～Ⅳ期患者中的阳性表达率分别为12.50%(1/8)、23.53%(4/17)、35.29%(6/17)和77.78%(7/9),CK20 mRNA在外周血中的阳性表达与TNM分期相关(P＜0.05);手术中有21例呈阳性表达(21/51),3例术前阴性表达者术中转为阳性表达,均为直肠癌手术患者;CK20 mRNA在外周血中的阳性表达与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值、肿瘤分化程度、浸润深度无关(P＞0.05),但与淋巴结转移、远处转移、TNM分期相关(P＜0.05);MVD与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值无关(P＞0.05),但与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关(P＜0.05);LMVD与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值、肿瘤分化程度、浸润深度无关(P＞0.05),但与淋巴结转移、远处转移、TNM分期相关(P＜0.01);大肠癌患者外周血CK20 mRNA阳性表达组MVD和LMVD的值分别为58.83±18.28和9.72±1.64,而外周血CK20mRNA阴性表达组MVD和LMVD的值分别为30.48±9.31和8.21±1.87,显著低于阳性表达组(P＜0.01)。
结论大肠癌细胞外周血微转移是大肠癌的早期事件,外周血微转移发生率随临床病理分期的增加而升高,外周血微转移与淋巴结转移、远处转移、TNM分期相关,是反映大肠癌生物学行为的指标之一;手术操作有可能促进直肠癌患者外周血微转移的发生;MVD与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关;LMVD与淋巴结转移、远处转移、TNM分期相关;大肠癌组织血管化程度和淋巴管生成与外周血微转移密切相关,通过对二者的检测有助于判断外周血微转移发生的情况。
第二部分Tiam1、DcR3在大肠癌中表达的临床意义及与大肠癌外周血微转移的关系
目的侵袭和转移是恶性肿瘤的重要特征,也是肿瘤患者死亡的主要原因,因此对肿瘤的侵袭和转移及其相关基因的研究具有重要意义。T淋巴瘤侵袭转移诱导因子1(T lymphoma invasion and metastasis factor 1,Tiam1)与肿瘤侵袭和转移密切相关。DcR3(decoy receptor 3)是肿瘤坏死因子受体超家族成员,在多种人类恶性肿瘤中出现过度表达,且DcR3的表达状态与恶性肿瘤的淋巴结转移和病理分期密切相关。关于Tiam1和DcR3与大肠癌外周血微转移的研究报道不多,我们通过检测Tiam1和DcR 3在大肠癌组织和正常大肠粘膜组织中的表达,探讨了Tiam1和DcR3表达与大肠癌临床病理参数的关系以及二者与大肠癌外周血微转移的关系。
方法以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,采用RT-PCR方法检测大肠癌患者外周血微转移;采用实时荧光定量RT-PCR方法检测大肠癌组织和正常组织中Tiam1和DcR3的表达;分析大肠癌组织中Tiam1和DcR3表达与临床病理参数以及患者外周血微转移的关系。
结果51例大肠癌组织及正常粘膜组织中均见Tiam1 mRNA表达,表达水平分别为(20.61±4.99)×10~(-4)和(3.49±2.74)×10~(-4),大肠癌组织中Tiam1mRNA表达明显高于正常组织(P＜0.01);Tiam1 mRNA在伴有外周血微转移的大肠癌组织中的表达水平为(72.11±2.38)×10~(-4),在不伴外周血微转移的大肠癌组织中的表达水平为(15.78±5.01)×10~(-4),二者差异有统计学意义(P＜0.05);大肠癌组织中Tiam1 mRNA表达水平与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值无关(P＞0.05),但与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关(P＜0.05);51例大肠癌组织及正常粘膜组织中均见DcR3 mRNA的表达,表达水平分别为(42.40±2.83)×10~(-4)和(12.59±2.55)×10~(-4),大肠癌组织中DcR3 mRNA表达明显高于正常组织(P＜0.01);DcR3 mRNA在伴有外周血微转移的大肠癌组织中表达水平为(61.53±2.60)×10~(-4),在不伴外周血微转移的大肠癌组织中表达水平为(34.60±2.84)×10~(-4),但二者差异无统计学意义(P＞0.05);DcR3 mRNA的表达水平与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值无关(P＞0.05),但与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关(P＜0.05)。
结论大肠癌组织的Tiam1 mRNA和DcR3 mRNA高表达与大肠癌的发生相关;大肠癌组织的Tiam1 mRNA和DcR3 mRNA表达水平与肿瘤分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关;Tiam1 mRNA高表达是大肠癌患者发生外周血微转移的高危因素;但DcR3 mRNA高表达不能作为大肠癌患者发生外周血微转移的高危因素。
第三部分Kallikrein6在大肠癌中表达的临床意义及与大肠癌外周血微转移的关系
目的激肽释放酶(Kallikreins,KLK)是一组分泌型丝氨酸蛋白酶,存在于多种组织和生物体液中。人KLK基因家族参与恶性肿瘤的发生发展,多种KLK与前列腺癌、卵巢癌、乳腺癌和睾丸癌等内分泌相关肿瘤的发生发展密切相关,是此类肿瘤的潜在标志物。KLK6异常表达与多种肿瘤相关。国内目前未见有关KLK6和大肠癌关系的报道。我们通过检测KLK6 mRNA在大肠癌组织和正常大肠粘膜组织中的表达,探讨了KLK6表达与大肠癌临床病理资料的关系及其与外周血微转移的关系。
方法以CK20 mRNA作为外周血中大肠癌细胞存在的特异性标志物,采用RT-PCR方法检测大肠癌患者外周血微转移;采用实时荧光定量RT-PCR方法检测大肠癌组织和正常组织中KLK6的表达;分析大肠癌组织中KLK6表达与临床病理参数以及患者外周血微转移的关系。
结果51例大肠癌组织及正常粘膜组织中均见KLK6 mRNA的表达。大肠癌组织中和正常粘膜组织中KLK6 mRNA表达水平分别为(33.11±2.34)×10~(-4)和(12.88±2.51)×10~(-4),有高度统计学差异(P＜0.01);伴有外周血微转移的大肠癌组织中和不伴外周血微转移的大肠癌组织中KLK6 mRNA的表达水平分别为(40.64±2.05)×10~(-4)和(30.06±2.45)×10~(-4),差异无统计学意义(P＞0.05);大肠癌组织的KLK6 mRNA表达与患者的性别、年龄、肿瘤原发部位、肿瘤大小、外周血CEA值、肿瘤分化程度、浸润深度、远处转移无关(P＞0.05),但与淋巴结转移、TNM分期相关(P＜0.05)。
结论大肠癌组织的KLK6 mRNA高表达与大肠癌的发生相关;大肠癌组织的KLK6 mRNA表达水平与淋巴结转移、TNM分期相关,与外周血微转移不相关,大肠癌组织的KLK6 mRNA的高表达不能作为大肠癌患者发生外周血微转移的高危因素。
Part One Clinical significance of peripheral blood micro-metastasis in colorectal cancer
Objective Peripheral blood micro-metastasis is one of the important factors for failure of surgical resection in colorectal cancer.Above 50%of the patients underwent surgical resection died of cancerous relapse or metastasis,indicating that micro-metastasis may have exited before the sugeries.CK20 mRNA has thought to be a specific biomarker for early diagnosis of micro-metastasis in colorectal cancer.To explore the role of determination of peripheral blood micro-metastasis in prognosis of colorectal cancer and to explore whether microvessel density(MVD) and lymphatic microvessel density(LMVD) have correlation with peripheral blood micro-metastasis.
Methods Using CK20 mRNA as a specific biomarker of tumor cells in peripheral blood,we determined its levels before and after surgical resection with the RT-PCR technique,also we determined the MVD and LMVD levels by immunohistochemistry.
Results Eighteen of the 51 patients were found positive CK20 mRNA expressions before surgical resection,of them,1(12.50%) was of the 8 patients with stageⅠ,4 (23.53%) was of the 17 patients with stageⅡ,6(35.29%) was of the 17 patients with stageⅢ,and 7(77.78%)was of the 9 patients with stageⅣ.These findings indicated that CK20 mRNA expression in peripheral blood correlated with the TNM classification in colorectal cancer.After surgical resection,3 more patients with rectum cancer were also shown positive CK20 mRNA expressions.CK20 mRNA expression had no relations to age,gender,primary location,tumor size,peripheral blood carcino-embryonic antigen level,and tumor differentiation and invasion,but had relations to lymphonode metastasis,distant metastasis and tumor TNM stage (P＜0.05).The MVD levels had no relations to age,gender,primary location,tumor size and the peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphonode metastasis,distant metastasis,tumor TNM stage(P＜0.05).Also the LMVD levels had no relations to age,gender,primary location,tumor size,the peripheral blood carcino-embryonic antigen level,and tumor differentiation and invasion,but had relations to lymphonode metastasis,distant metastasis,and tumor TNM stage(P＜0.05).The MVD and LMVD levels were 58.83±18.28 and 9.72±1.64 in the positive CK20 mRNA group,however they were 30.48±9.31 and 8.21±1.87 in the negative CK20 mRNA group,the difference was statistically significant(P＜0.01 ).
Conclusions Peripheral blood micro-metastasis is related to lymphnode metastasis, distant metastatis and TNM stage.Surgical procedures may promote the genesis of periphral blood micro-metastasis.Determination of tissue vascularization and genesis of lymphatic vessel is helpful in judging genesis of peripheral blood micro-metastasis in colorectal cancer.
Part Two Tiam1 and DcR3 expressions in colorectal cancer and their correlation with peripheral blood micro-metastasis
Objective Invasion and metastasis are key characteristics of malignant tumors.It has been shown that T lymphoma invasion and metastasis factor 1(Tiam1) closely correlates with tumor invasion and metastasis.Decoy receptor 3(DcR3) is one member of the tumor necrosin receptor hyperfamily,and it is overexpressed in many human malignant tumors,also its overexpression closely relates to tumor lymphnode metastasis and pathologic staging.To determine the expressions of Tiam1 mRNA and DcR3 mRNA in cancerous tissues and non-cancerous tissues of colorectal cancer,and to explore their correlation with pathologic data and peripheral blood micro-metastasis.
Methods Using the RT-PCR method,the CK20 mRNA expression was determined.Using fluorescence quantitative-RT-PCR(FQ-RT-PCR),the expressions of Tiam1 mRNA and DcR3 mRNA were determined in 51 cases of colorectal carcinoma.
Results The Tiam1 mRNA expression level was(20.61±4.99)×10~(-4) in cancerous tissues and was(3.49±2.74)×10~(-4) in non-cancerous tissues(P＜0.01).Also it was (72.11±2.38)×10~(-4) in cancerous tissues with micro-metastasis and was(15.78±5.01)×10~(-4) in cancerous tissues without micro-metastasis,the difference was statistically significant(P＜0.05).Tiam1 mRNA expression had no relations to age,gender, primary location,tumor size and the peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphnode metastasis, distant metastasis and tumor TNM stage(P＜0.05).The DcR3 mRNA expression level was(42.40±2.83)×10~(-4) in cancerous tissues and was(12.59±2.55)×10~(-4) in non-cancerous tissues(P＜0.01).Also it was(61.53±2.60)×10~(-4) in cancerous tissues with micro-metastasis and was(34.60±2.84)×10~(-4) in cancerous tissues without micro-metastasis,but the difference was not statistically significant(P＞0.05).DcR3 mRNA expression had no relations to age,gender,primary location,tumor size and peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphnode metastasis,distant metastasis and tumor TNM stage(P＜0.05).
Conclusions Overexpressions of Tiam1 mRNA and DcR3 mRNA correlate with genesis of colorectal cancer.Tiam1 mRNA overexpression is a high risk factor for peripheral blood micro-metastasis in colorectal cancer,but DcR3 mRNA can not be used as a high risk factor for peripheral blood micro-metastasis in colorectal cancer.
Part Three Human KaUikrein6 expression in colorectal cancer and its correlation with peripheral blood micro-metastasis
Objective Kallikreins(KLKs) have a similar genomic organization and show significant homology at both the nucleotide and the protein level.Human KLK gene family has participated in genesis and development of cancers.A growing body of evidence suggests that KLKs are involved in human malignancies and that many KLKs are promising biomarkers of prostate,ovarian,breast and testicular cancers.To determine the expression of KLK6 mRNA in colorectal cancer and to explore its relationship to clinicopathologic features and peripheral blood micro-metastasis in colorectal cancer.
Methods Using the RT-PCR method,the CK20 mRNA expression was determined.Using the FQ-RT-PCR method,the KLK6 mRNA expression was determined in 51 cases of colorectal carcinoma.
Results The KLK6 mRNA expression level was(33.11±2.34)×10~(-4) in cancerous tissues and was(12.88±2.51)×10~(-4) in non-cancerous tissues(P＜0.01).Also it was (40.64±2.05)×10~(-4) in cancerous tissues with micro-metastasis and was(30.06±2.45)×10~(-4) in cancerous tissues without micro-metastasis,the difference was not statistically significant(P＞0.05).KLK6 mRNA expression had no relations to age, gender,primary location,tumor size,peripheral blood carcino-embryonic antigen level,tumor differentiation and invasion,and distant metastasis,but had relations to lymphnode metastasis and tumor TNM stage(P＜0.05).
Conclusions Overexpression of KLK6 mRNA correlates with genesis of colorectal cancer,however,it can not be used as a high risk factor of peripheral blood micro-metastasis in colorectal cancer.
引文
1.Parkin DM,Bray F,Ferlay J,et al.Global cancer statistics,2002,CA Cancer J Clin,2005,55:74-108.
2.周晓东,吕农华.大肠癌的流行病学研究现状.现代消化及介入诊疗,2006,11:149-151.
3.Weiss L,Grundmann E,Torhorst J,et al.Haematogenous metastatic patterns in colonic carcinoma:an analysis of 1541 necropsies.J Pathol,1986,150:195-203.
4.Penland SK,Goldberg RM.Current strategies in previously untreated advanced colorectal cancer.Oncology,2004,18:715-722.
5.Midgley RS,Kerr DJ.ABC of colorectal cancer:adjuvant therapy.BMJ,2000,321:1208-1211.
6.Hayashi N,Ito I,Yanagisawa A,et al.Genetic diagosis of lymphnode metastasis in colorectal cancer.Lancet,1995,345:1257-1297.
7.Burchill SA,Bradbury FM,Pittman K,et al.Detection of epithelial cancer cells in peripheral blood by reverse transcriptase polymerase chain reaction.Br J Cancer,1995,71:278-281.
8.Aihara T,Fujiwara Y,Miyake Y,et al.Mammaglobin B gene as a novel marker for lymphnode micrometastasisin patients with abdominal cancer.Cancer Lett,2000,150:79-84.
9.Burchill SA,Bradbury FM,Smith B,et al.Neuroblastoma cell detection by reverse transcriptase-polymerase chain reaction(RT-PCR) for tyrosine hydroxylase mRNA.Int J Cancer,1994,57:671-675.
10.蔡刚,李闻捷,沈茜.实时逆转录聚合酶链反应绝对定量实验优化的研究.上海医学检验杂志,2003,18:343-346.
11.Rasmussen R.Quantification on the light cycler.Rapid cycle real-time PCR,methods and applications.Heidelberg:Springer Press,2001:21.
12.蔡文琴,王伯云.实用免疫细胞化学与核酸分子杂交技术.成都:四川科学技术出版社,1994:72-89.
13. Weidner N. Intratumor microvessel density as a prognostic factor in cancer. Am J Pathol,1995,147:9-19.
14. Ohno M,Nakamura T , Kunimoto Y, et al . Lymphagenesis correlates with expression of vascular endothelial growth factor-C in Colorectal cancer. Oncol Rep, 2003,10:939-943.
15. Mitas M, Hoover L, Silvestri G, et al. Lunx is a superior molecular marker for detection of non-small cell lung cancer in peripheral blood. J Mol Diagn, 2003, 5: 237-242.
16. Gure AO , Chua R , Williamson B , et al. Cancer-testis genes are coordinately expressed and are markers of poor outcome in non-small cell lung cancer . Clin Cancer Res, 2005, 11: 8055-8062.
17. Cheung ST, Leung KL, Ip YC, et al. Claudin210 expression level is associated with recurrence of primary hepatocellular carcinoma. Clin Cancer Res, 2005, 11: 551-556.
18. Poon RT, Chung KK, Cheung ST, et al. Clinical significance of thrombospondin 1 expression in hepatocellular carcinoma. Clin Cancer Res, 2004, 10: 4150-4157.
19. Sonoda H, Yamamoto K, Kushima R, et al. Detection of lymphnode micrometastasis in gastric cancer by MUC2 RT-PCR: usefulness in pT1 cases. J Surg Oncol, 2004, 88: 63-70.
20. Shimomura K, Sakakura C, Takemura M, et al. Combination of L-3-phosphoserine phosphatase and CEA using real-time RT-PCR improves accuracy in detection of peritoneal micrometastasis of gastric cancer. Anticancer Res, 2004, 24: 1113-1120.
21. Baker MK, Mikhitarian K, Osta W, et al. Molecular detection of breast cancer cells in the peripheral blood of advanced stage breast cancer patients using multimarker real-time reverse transcription-poly-merase chain reaction and a novel porous barrier density gradient centrifugation technology. Clin Cancer Res, 2003, 9: 4865-4871.
22. Masuda TA , Kataoka A , Ohno S, et al. Detection of occult cancer cells in peripheral blood and bone marrow by quantitative RT-PCR assay for cytokeratin-7 in breast cancer patients. Int J Oncol, 2005, 26: 721-730.
23. Patel K, Whelan PJ, Prescott S, et al. The use of real-time reverse transcription-PCR for prostate-specific antigen mRNA to discriminate between blood samples from healthy volunteers and from patients wit h metastatic prostate cancer. Clin Cancer Res, 2004,10:7511-7519.
24. Hara N, Kasahara T, Kawasaki T, et al. Reverse transcription-polymerase chain reaction detection of prostate-specific antigen, prostate-specific membrane antigen, and prostate stem cell antigen in one milliliter of peripheral blood: value for the staging of prostate cancer. Clin Cancer Res, 2002, 8:1794-1799.
25. Buskens CJ , Van Rees BP , Sivula A , et al. Prognostic significance of elevated cyclooxygenase 2 expression in patients with adenocarcinoma of the esophagus. Gastroenterology, 2002, 122: 1800-1807.
26. Sakakibara T, Hibi K, Kodera Y, et al. Plasminogen activator inhibitor-1 as a potential marker for the malignancy of esophageal squamous cell carcinoma. Clin Cancer Res, 2004, 10: 1375 - 1378.
27. Ribuffo D, Gradilone A, Vonella M, et al. Prognostic significance of reverse transcriptase-polymerase chain reaction negative sentinel nodes in malignant melanoma. Ann Surg Oncol, 2003,10: 396-402.
28. Scala S, Ottaiano A, Ascierto PA, et al. Expression of CXCR4 predicts poor prognosis in patients with malignant melanoma. Clin Cancer Res, 2005, 11: 1835-1841.
29. Iinuma H, Okinaga K, Egami H, et al. Usefulness and clinical significance of quantitative real-time RT-PCR to detect isolated tumor cells in the peripheral blood and tumor drainage blood of patients with Colorectal cancer. Int J Oncol, 2006, 28: 297-306.
30. Hernandez BY, Frierson HF, Moskaluk CA, et al. CK20 and CK7 protein expression in Colorectal cancer: demonstration of the utility of a population-based tissue microarray.Hum Pathol, 2005, 36: 275-281.
31. Dandachi N, Balic M, Stanzer S, et al .Critical evaluation of real-time reverse transcriptasepolymerase chain reaction for the quantitative detection of cytokeraitn
20 mRNA incolorectal cancer paitents.J Mol Diagn,2005,7:631-637.
32.苏琪,韩霞,殷红专.CK20诊断大肠癌微转移的价值.世界华人消化杂志,2004,12:2153-2155.
33.徐青,陈瑞新,王志伟,等.大肠癌组织血管生成与患者外周血CK20 mRNA 表达的临床研究.中华医学杂志,2005,85:1205-1208.
34.袁正强,曹建林,陈微微,等.大肠癌患者外周血癌细胞CK20检测及其意义.临床肿瘤学杂志,2005,10:74-76.
35.Berois N,Varangot M,Aizen B,et al.Molecular detection of cancer cells in bone marrow and peripheral blood of patients with operable breast cancer,Comparison of ckl9,Muci and CEA using RT-PCR.European J Cancer,2000,36:717-723.
36.Oberg AN,Lindmark GE,Israelsson AC,et al.Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS.Int J Cancer,2004,111:101-110.
37.Moll R,Schiller DL,Franke WW,et al.Identification of protein IT of the intestinal cytoskeleton as a novel type Ⅰ cytokeratin with unusual properties and expression pattenrs.J Cell Biol,1990,111:567-580.
38.Moll R,Lowe A,Laufer J,et al.Cytokeratin 20 in human carcinomas.A new histodiagnostic marker detected by monoclonal antibodies.Am J Pathol,1992;140:427-447.
39.Aihara T,Fujiwara Y,Miyake Y,et al.Mammaglobin B gene as a novel marker for lymphnode micrometastasisin patients with abdominal cancer.Cancer Lett,2000,150:79-84.
40.陈英剑.荧光实时逆转录PCR定量研究进展.国外医学临床生物化学与检验学分册,2004,25:348-351.
41.Bustin SA.Quantification of mRNA using real-time reverse transcription PCR (RT-PCR):trends and problems.J Mol Endocrinol,2002,29:23-39.
42.陈梓宏,梁荣,赖振南,等.大肠癌患者外周血CEAmRNA和CK20mRNA 的联合检测及临床意义.四川肿瘤防治,2007,20:100-102.
43.易秉强,肖白,吴燕.实时定量RT-PCR检测大肠癌病人外周血CK20mRNA 和CEAmRNA的研究.中华腹部疾病杂志,2005,5:391-394.
44.Zhang XW,Yang HY,Fan P,et al.Detection of micrometastasis in peripheral blood by multi-sampling in patients with colorectal cancer.World J Gastroenterol,2005,11:436-438.
45.Nishizaki T,Matsumata T,Kanematsu T,et al.Surgical manipulation of VX2carcinoma in the rabbit liver evokes enhancement of metastasis.J Surg Res,1990,49:92-97.
46.Garcia-Olmo D,Ontanon J,Garcia-Olmo DC,et al.Experimental evidence does not support use of the "no-touch" isolation technique in colorectal cancer.Dis Colon Rectum,1999,42:1449-1456.
47.Yamaguchi K,Takagi Y,Aoki S,et al.Significant detection of circulating cancer cells in the blood by reverse transcriptase-polymerase chain reaction during colorectal cancer resection.Ann Surg,2000,232:58-65.
48.Weitz J,Koch M,Kienle P,et al.Detection of hematogenic tumor cell dissemination in patients undergoing resection of liver metastases of colorectal cancer.Ann Surg,2000,232:66-72.
49 Weitz J,Kienle P,Magener A,et al.Detection of disseminated colorectal cancer cells in lymph nodes,blood and bone marrow.Clin Cancer Res,1999,5:1830-1836.
50.林国乐,邱辉忠,徐彤,等.大肠癌患者术前和术中外周血微转移的检测及其临床意义.中华普通外科杂志,2002:17:605-607.
51.Dalal S,Berry AM,Cullinane CJ,et al.Vascular endothelial growth factor:a therapeutic target for tumors of the Ewing's sarcoma family.Clin Cancer Res,2005,11:2364-2378.
52.Boxer GM,Tsiompanou E,Levine T,et al.Immunohistochemical expression of vascular endothelial growth factor and microvessel counting as prognostic indicators in node-negative colorectal cancer.Tumour Biol,2005,26:1-8.
53.Uzzan B,Nicolas P,Cucherat M,et al.Microvessel density as a prognostic factor in women wit h breast cancer:a systematic review of the literature and meta-analysis.Cancer Res,2004,64:2941-2955.
54.Giatromanolaki A.Prognostic role of angiogenesis in non-small cell lung cancer.Anticancer Res,2001,21:4373-4382.
55.Sonmezer M,Gungor M,Ensari A,et al.Prognostic significance of tumor angiogenesis in epithelial ovarian cancer:in association with transforming growth factor beta and vascular endothelial growth factor.Int J Gynecol Cancer,2004,14:82-88.
56.Witte MH,Bernas M J,Martin CP,et al.Lymphangiogenesis and lymphangiodysplasia:from molecular to clinical lymphology.Microsc Res Tech,2001,55:122-145.
57.Niakosari F,Kahn HJ,Marks A,et al.Detection of lymphatic invasion in primary melanoma with monoclonal antibody D2-40:a new selective immunohistochemical marker of lymphatic endothelium.Arch Dermatol,2005,141:440-444.
58.Shields JD,Borsetti M,Rigby H,et al.Lymphatic density and metastatic spread in human malignant melanoma.Br J Cancer,2004,90:693-700.
59.牟江洪,阎晓初,李增鹏,等.结直肠癌淋巴管生成的特点及其临床病理意义.中华病理学杂志,2005,34:348-352.
60.阎晓初,牟江红,柳凤轩.大肠癌组织淋巴管分布特点及其与转移和预后的关系.第三军医大学学报,2005,27:2255-2258.
61.姜金波,朱民,李雪梅,等.结肠癌VEGF-C表达、淋巴管密度与淋巴转移和预后的关系.山东大学学报(医学版),2005,43:616-620.
1.周晓东,吕农华.大肠癌的流行病学研究现状.现代消化及介入诊疗,2006,11:149-151.
2.Midgley RS,Kerr DJ.ABC of colorectal cancer:adjuvant therapy.BMJ,2000,321:1208-1211.
3.Habets GG,Scholtes EH,Zuydgeest D.Identification of an invasion-inducing gene,Tiam-1,that encodesa protein with homology to GDP-GTP ecchangers for Rho-like protein.Cell,1994,77:537-549.
4.Mertens AE,Roovers RC,Collard JG.Regulation of Tiaml-Rac signalling.FEBS Lett,2003,546:11-16.
5.Minard ME,Kim LS,Price JE,et al.The role of the guanine nucleotide exchange factor Tiam 1 in cellular migration,invasion,adhesion and tumor progression.Breast Cancer Res Treat,2004,84:21-32.
6.Bourguignon LY,Zhu H,Shao L,et al.Ankyrin-Tiaml interaction promotes Racl signaling and metastatic breast tumor cell invasion and migration.J Cell Biol,2000,150:177-191.
7.Malliri A,van der Kammen RA,Clark K,et al.Mice deficient in the Rac activator Tiam 1 are resistant to Ras-induced skin tumours.Nature,2002,417:867-871.
8.钟大平,周进明,章容,等.Tiam1和Rac1在大肠癌组织中的表达及其临床意义.中华肿瘤防治杂志,2007,14:129-132.
9.Minard ME,Herynk MH,Collard JG,et al.The guanine nucleotide exchange factor Tiaml increases colon carcinoma growth at metastatic sites in an orthotopic nude mouse model.Oncogene,2005,24:2568-2573.
10.Pitti RM,Marsters SA,Lawrence DA,et al.Genomic amplification of a decoy receptor for Fas ligand in lung and colon cancer.Nature,1998,396:699-703.
11.Bai C,Connolly B,Metzker ML,et al.Overexpression of M68/DcR3 in human gastrointestinal tract tumors independent of gene amplification and its location in a four-gene cluster.Proc Natl Acad Sci U S A,2000,97:1230-1235.
12.Yu KY,Kwon B,Ni J,et al.A newly identified member of tumor necrosis factor receptor superfamily(TR6) suppresses LIGHT-mediated apoptosis.J Biol Chem,1999,274:13733-13736.
13.Wu Y,Han B,Sheng H,et al.Clinical significance of detecting elevated serum DcR3/TR6/M68 in malignant tumor patients.Int J Cancer,2003,105:724-732.
14.Takahama Y,Yamada Y,Emoto K,et al.The prognostic significance of overexpression of the decoy receptor for Fas ligand(DcR3) in patients with gastric carcinomas.Gastric Cancer,2002,5:61-68.
15.卢佩颖,张红河.TIAM1与KAI1基因在胃癌组织中的表达研究.浙江临床医学,2005,7:725.
16.黄志为,马忠森.DcR3基因检测在恶性肿瘤研究中的地位.广西医科大学学报,2004,21:182-184.
17.Burchill SA,Bradbury MF,Pittman K,et al.Detection of epithelial cancer cells in peripheral blood by reverse transcriptase polymerase chain reaction.Br J Cancer,1995,71:278-281.
18.蔡刚,李闻捷,沈茜.实时逆转录聚合酶链反应绝对定量实验优化的研究.上海医学检验杂志,2003,18:343-346.
19.Rasmussen R.Quantification on the light cycler.Rapid cycle real-time PCR,methods and applications.Heidelberg:Springer Press,2001:21.
20.Zheng Y.Db1 family guanine nucleotide exchange factors.Trends Biochem Sci,2001,26:724-732.
21.Robbe K,Otto-Bruc A,Chardin P,et al.Dissociation of GDP dissociation inhibitor and membrane translocation are required for efficient activation of Rac by the Db1 homology region of Tiam.J Biol Chem,2003,278:4756-4762.
22.Crompton AM,Foley LH,Wood A,et al.Regulation of Tiaml nucleotide exchange activity by pleckstrin domain binding ligands.J Biol Chem,2000,275:25751-25759.
23.Stam JC,Sander EE,Michiels F,et al.Targeting of Tiaml to the plasma membrane requires the cooperative function of the N-terminal pleckstrin homology domain and an adjacent protein interaction domain.J Biol Chem,1997,272:28447 -28454.
24.李新元,张建中,王登顺,等.肿瘤侵袭诱导基因蛋白在正常及癌组织中的表达及意义.中华病理学杂志,2000,29:294-295
25.Malliri A,Colard JG.Role of Rho-family proteins in cell adhesion and cancer.Curr Opin Cell Biol,2003,15:583-589.
26.Schmidt A,Hall A.Guanine nucleotide exchange factors for Rho GTPases:turning on the switch.Genes Dev,2002,16:1587-1609.
27.Fleming IN,Elilott CM,Buchanan FG,et al.Ca~(2+)/calmodulin-dependent protein kinase Ⅱ regulates Tiam 1 by reversible portein phosphorylation.J Biol Chem,1999,274:12753-12758.
28.Lambert JM,Lambert QT,Reuther GW,et al.Tiaml mediates Ras activation of Rac by a PI(3) K-independent mechanism.Nat Cell Biol,2002,4:621-625.
29.Buchsbaum RJ,ConnollyBA,FeigLA.Interaction of Rac exchange factors Tiaml and Ras-GRF1 with a scaffold for the p38 mitogen-activated protein kinase cascade.Mol Cell Biol,2002,22:4073-4085.
30.Matsuo N,Terao M,Nabeshima Y,et al.Roles of STEF/Tiaml,guanine nucleotide exchange factors for Racl,in regulation of growth cone morphology.Mol Cell Neurosci,2003,24:69-81.
31.Kawauchi T,Chihama K,Nabeshima Y,et al.The in vivo roles of STEF/Tiaml,Racl and JNK in cortical neuronal migration.EMBO J,2003,22:4190-4201.
32.Kunda P,Paglini G,Quiroga S,et al.Evidence for the involvement of Tiaml in axon formation.J Neurosci,2001,21:2361-2372.
33.van Leeuwen FN,van der Kammen RA,Habets GG,et al.Oncogenic activity of Tiam 1 and Racl in NIH3T3 cells.Oncogene,1995,11:2215-2221.
34.Bourguignon LY,Zhu H,Shao L,et al.CD44 interaction with tiaml promotes Racl signaling and hyaluronic acid-mediated breast tumor cell migration.J Biol Chem,2000,275:1829-1838.
35.朱金明,余佩武.T淋巴瘤侵袭转移诱导因子1在胃癌组织中的表达及其临床意义.现代肿瘤医学,2005,13:320-323.
36.朱金明,余佩武,赵永亮.Tiam 1与胃癌细胞侵袭转移及与裸鼠移植瘤形成的关系.中国普通外科杂志,2007,16:855-858.
37.吴明富,史艳燕,韩志强,等.人卵巢癌组织中Tiam 1和Rac 1的表达与肿瘤侵袭转移的关系.中国肿瘤临床,2005,32:910-914.
38.莫立根,王会河,黄光武,等.Tiam 1基因表达与鼻咽癌浸润转移的关系.临床耳鼻咽喉科杂志,2005,19:785-787.
39.刘莉,许岸高,杨玉芳,等.Tiam 1对结直肠癌细胞生物学特性的影响.中华病理学杂志,2005,34:664-668.
40.Engers R,Springer E,Michiels F,et al.Rac affects invasion of human renal cell carcinomas by up-regulating tissue inhibitor of metalloproteinases(TIMP)-1 and TIMP-2 expression.J Biol Chem,2001,276:41889-41897.
41.Hordijk PL,ten Klooster JP,van der Kanunen RA,et al.Inhibition of invasion of epithehal cells by Tiaml-Rac signaling.Science,1997,278:1464-1466.
42.Servitja JM,Marinissen MJ,Sodhi A,et al.Rac 1 function is required for Src-induced transformation:evidence of a role for Tiara 1 and Vav2 in Rac aetivation by Src.J Biol Chem,2003,278:34339-34346.
43.Bourguignon LY,Zhu H,Shao L,et al.Ankyrin-Tiaml interaction promotes Racl signaling and metastatic breast tumor cell invasion and migration.J Cell Biol,2000,150:177-191.
44.Otsuld Y,Tanaka M,Kanlo T,et al.Guanine nucleotide exchange factor,Tiam1,directly binds to c-Myc and interferes with c-Myc-mediated apoptosis in rat-1 fibroblasts.J Biol Chem,2003,278:5132-5140.
45.Bourguignon LY.CD44-mediated oncogenic signaling and cytoskeleton activation during mammary tumor progression.J Mammary GIand Biol Neoplasia,2001,6:287-297.
46.Li H,Zhang L,Lou H,et al.Overexpression of decoy receptor 3 in precancerous lesions and adenocarcinoma of the esophagus.Am J Clin Pathol,2005,124:282-287.
47.Chen J,Zhang L,Kim S.Quantification and detection of DcR3,a decoy receptor in TNFR family.J Immunol Methods,2004,285:63-70.
48. Arakawa Y, Tachibana O, Hasegawa M, et al. Frequent gene amplification and overexpression of decoy receptor 3 in glioblastoma. Acta Neuropathol, 2005, 109: 294-298.
49. Ohshima K, Haraoka S, Sugihara M, et al. Amplification and expression of a decoy receptor for fas ligand (DcR3) in virus (EBV or HTLV-I) associated lymphomas. Cancer Lett, 2000, 160: 89-97.
50. Tsuji S, Hosotani R, Yonehara S, et al. Endogenous decoy receptor 3 blocks the growth inhibition signals mediated by Fas ligand in human pancreatic adenocarcinoma. Int J Cancer, 2003, 106:17-25.
51. Shen HW, Gao SL, Wu YL, et al. Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis. World J Gastroenterol, 2005, 11: 5926-5930.
52. Maeda T, Hao C, Tron VA. Ultraviolet light (UV) regulation of the TNF family decoy receptors DcR2 and DcR3 in human keratinocytes. J Cutan Med Surg, 2001, 5: 294-298.
53. Mild G, Bachmann F, Boulay JL, et al. DCR3 locus is a predictive marker for 5-fluorouracil-based adjuvant chemotherapy in Colorectal cancer. Int J Cancer, 2002, 102: 254-257.
1. Clements J, Hooper J, Dong Y, et al. The expanded human kallikrein (KLK) gene family: genomic organisation, tissue-specific expression and potential functions. Biol Chem, 2001, 382: 5-14.
2. Diamandis EP, Yousef GM, Clements J, et al. New nomenclature for the human tissue kallikrein gene family. Clin Chem, 2000,46: 1855-1858.
3. Yousef GM, Obiezu CV, Luo LY, et al. Human tissue kallikreins: from gene structure to function and clinical applications. Adv Clin Chem, 2005,39:11-79.
4. Yousef GM, Diamandis EP. The new human tissue kallikrein gene family: structure, function, and association to disease. Endocr Rev, 2001,22: 184-204.
5. Borgono CA, Diamandis EP. The emerging roles of human tissue kallikreins in cancer. Nat Rev Cancer, 2004,4: 876-890.
6. Borgono CA, Michael IP, Diamandis EP. Human tissue kallikreins: physiologic roles and applications in cancer. Mol Cancer Res, 2004,2: 257-280.
7. Diamandis EP. Prostate-specific antigen: its usefulness in clinical medicine. Trends Endocrinol Metab, 1998, 9: 310-316.
8. Anisowicz A, Sotiropoulou G, Stenman G, et al. A novel protease homolog differentially expressed in breast and ovarian cancer. Mol Med, 1996, 2: 624-636.
9.Yamashiro K, Tsuruoka N, Kodama S, et al. Molecular cloning of a novel trypsin-like serine protease (neurosin) preferentially expressed in brain. Biochim BiophysActa, 1997,1350: 11-14.
10. Little SP, Dixon EP, Norris F, et al. Zyme, a novel and potentially amyloidogenic enzyme cDNA isolated from Alzheimer's disease brain. J Biol Chem, 1997, 272: 25135-25142.
11. Diamandis EP, Scorilas A, Fracchioli S, et al. Human kallikrein 6 (hK6): a new potential serum biomarker for diagnosis and prognosis of ovarian carcinoma. J Clin Oncol, 2003, 21: 1035-1043.
12. Hoffman BR, Katsaros D, Scorilas A, et al. Immunofluorometric quantitation and histochemical localisation of kallikrein 6 protein in ovarian cancer tissue:a new independent unfavourable prognostic biomarker.Br J Cancer,2002,87:763-771.
13.Tanimoto H,Underwood LJ,Shigemasa K,et al.Increased expression of protease M in ovarian tumors.Tumour Biol,2001,22:11-18.
14.Wolf WC,Evans DM,Chao L,et al.A synthetic tissue kallikrein inhibitor suppresses cancer cell invasiveness.Am J Pathol,2001,159:1797-1805.
15.Diamandis EP,Yousef GM,Soosaipillai AR,et al.Immunofluorometric assay of human kallikrein 6(zyme/protease M/neurosin) and preliminary clinical applications.Clin Biochem,2000,33:369-375.
16.Yousef GM,Borgono CA,Popalis C,et al.In-silico analysis of kallikrein gene expression in pancreatic and colon cancers.Anticancer Res,2004,24:43-51.
17.Ogawa K,Utsunomiya T,Mimori K,et al.Clinical significance of human kallikrein gene 6 messenger RNA expression in colorectal cancer.Clin Cancer Res,2005,11:2889-2893.
18.Ghosh MC,Grass L,Soosaipillai A,et al.Human kallikrein 6 degrades extracellular matrix proteins and may enhance the metastatic potential of tumour cells.Tumour Biol,2004,25:193-199.
19.Iwata A,Maruyama M,Akagi T,et al.Alpha-synuclein degradation by serine protease neurosin:implication for pathogenesis of synucleinopathies.Hum Mol Genet,2003,12:2625-2635.
20.Burchill SA,Bradbury MF,Pittman K,et al.Detection of epithelial cancer cells in peripheral blood by reverse transcriptase polymerase chain reaction.Br J Cancer,1995,71:278-281.
21.蔡刚,李闻捷,沈茜.实时逆转录聚合酶链反应绝对定量实验优化的研究.上海医学检验杂志,2003,18:343-346.
22.Rasmussen R.Quantification on the light cycler.Rapid cycle real-time PCR,methods and applications.Heidelberg:Springer Press,2001:21.
23.Schmaier AH.The plasma kallikrein-kinin system counterbalances the renin-angiotensin system.J Clin Invest,2002,109:1007-1009.
24.Diamandis EP,Yousef GM,Luo LY,et al.The new human kallikrein gene family:implications in carcinogenesis.Trends Endocrinol Metab,2000,11:54-60.
25.Yu H,Levesque MA,Clark GM,et al.Prognostic value of prostate-specific antigen for women with breast cancer:A large United States cohort study.Clin Cancer Res,1998,4:1489-1497.
26.Yousef GM,Diamandis EP.Human kallilreins:common structural features,sequence analysis and evaluation.Curr Genomics,2003,276:127-232.
27.Clements JA,Willemsen NM,Myers SA,et al.The tissue kallikrein family of serine proteases:functional roles in human disease and potential as clinical biomarkers.Crit Rev Clin Lab Sci,2004,41:265-312.
28.Jin E,Fujiwara M,Pan X,et al.Protease-activated receptor(PAR)-1 and PAR-2participate in the cell growth of alveolar capillary endothelium in primary lung adenocarcinomas.Cancer,2003,97:703-713.
29.Schuster R,Max N,Mann B,et al.Quantitative real-time RT-PCR for detection of disseminated tumor cells in peripheral blood of patients with colorectal cancer using different mRNA markers.Int J Cancer,2004,108:219-227.
30.Lu KH,Patterson AP,Wang L,et al.Selection of potential markers for epithelial ovarian cancer with gene expression arrays and recursive descent partition analysis.Clin Cancer Res,2004,10:3291-3300.
31.Ni X,Zhang W,Huang KC,et al.Characterisation of human kallikrein6/protease M expression in ovarian cancer.Br J Cancer,2004,91:725-731.
32.胡成进,陈英剑,闻新棉,等.KLK6 mRNA在乳腺癌中的表达及其临床病理和生物学意义.解放军医学杂志,2007,32:1085-1087.
33.Petraki CD,Gregorakis AK,Vaslamatzis MM,et al.Prognostic implications of the immunohistochemical expression of human kallikreins 5,6,10 and 11 in renal cell carcinoma.Tumour Biol,2006,27:1-7.
34.Nagahara H,Mimori K,Utsunomiya T,et al.Clinicopathologic and biological significance of kallikrein 6 overexpression in human gastric cancer.Clin Cancer Res,2005,11:6800-6806.
35.Liotta LA,Steeg PS,Stetler-Stevenson WG.Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation.Cell,1991,64:327-336.
36.Tryggvason K,Hoyhtya M,Salo T.Proteolytic degradation of extracellular matrix in tumor invasion.Biochim Biophys Acta,1987,907:191-217.
37.Duffy MJ.The role of proteolytic enzymes in cancer invasion and metastasis.Clin Exp Metastasis,1992,10:145-155.
38.Aznavoorian S,Murphy AN,Stetler-Stevenson WG,et al.Molecular aspects of tumor cell invasion and metastasis.Cancer,1993,71:1368-1383.
39.Matrisian LM.Cancer biology:extracellular proteinases in malignancy.Curr Biol,1999,9:R776-778.
40.Nelson AR,Fingleton B,Rothenberg ML,et al.Matrix metalloproteinases:biologic activity and clinical implications.J Clin Oncol,2000,18:1135-1149.
41.McCawley LJ,Matrisian LM.Tumor progression:defining the soil round the tumor seed.Curr Biol,2001,11:R25-27.
42.Magklara A,Mellati AA,Wasney GA,et al.Characterization of the enzymatic activity of human kallikrein 6:Autoactivation,substrate specificity,and regulation by inhibitors.Biochem Biophys Res Commun,2003,307:948-955.
43.Bernett MJ,Blaber SI,Scarisbrick IA,et al.Crystal structure and biochemical characterization of human kallikrein 6 reveals that a trypsin-like kallikrein is expressed in the central nervous system.J Biol Chem,2002,277:24562-24570.
44.Yousef GM,Luo LY,Scherer SW,et al.Molecular characterization of zyme/protease M/neurosin(PRSS9),a hormonally regulated kallikrein-like serine protease.Genomics,1999,62:251-259.
45.Yousef GM,Diamandis EP.Kallikreins,steroid hormones and ovarian cancer:is there a link? Minerva Endocrinol,2002,27:157-166.
46.张芳,陈英剑,胡成进.17β雌二醇对人卵巢癌细胞株H08910 KLK6基因表达的调节.山东大学学报(医学版),2007,45:42-45.
47.陈英剑,胡成进,闻新棉,等.雌激素对人乳腺癌细胞株MCF-7 KLK6mRNA和蛋白表达水平的影响.中国病理生理杂志,2007,23:246-249.
48.Qiu Y,Langman MJ,Eggo MC.Targets of 17 beta-oestradiol-induced apoptosis in colon cancer cells:a mechanism for the protective effects of hormone replacement therapy? J Endocrinol, 2004, 181: 327-337.
49. Campbell-Thompson M, Lynch IJ, Bhardwaj B. Expression of estrogen receptor (ER) subtypes and ERbeta isoforms in colon cancer. Cancer Res, 2001,61: 632-640.
50. Konstantinopoulos PA, Kominea A, Vandoros G, et al. Oestrogen receptor beta (ERbeta) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumour's dedifferentiation. Eur J Cancer, 2003, 39: 1251-1258.
2.周晓东,吕农华.大肠癌的流行病学研究现状.现代消化及介入诊疗,2006,11:149-151.
3.Weiss L,Grundmann E,Torhorst J,et al.Haematogenous metastatic patterns in colonic carcinoma:an analysis of 1541 necropsies.J Pathol,1986,150:195-203.
4.Penland SK,Goldberg RM.Current strategies in previously untreated advanced colorectal cancer.Oncology,2004,18:715-722.
5.Midgley RS,Kerr DJ.ABC of colorectal cancer:adjuvant therapy.BMJ,2000,321:1208-1211.
6.Hayashi N,Ito I,Yanagisawa A,et al.Genetic diagosis of lymphnode metastasis in colorectal cancer.Lancet,1995,345:1257-1297.
7.Burchill SA,Bradbury FM,Pittman K,et al.Detection of epithelial cancer cells in peripheral blood by reverse transcriptase polymerase chain reaction.Br J Cancer,1995,71:278-281.
8.Aihara T,Fujiwara Y,Miyake Y,et al.Mammaglobin B gene as a novel marker for lymphnode micrometastasisin patients with abdominal cancer.Cancer Lett,2000,150:79-84.
9.Burchill SA,Bradbury FM,Smith B,et al.Neuroblastoma cell detection by reverse transcriptase-polymerase chain reaction(RT-PCR) for tyrosine hydroxylase mRNA.Int J Cancer,1994,57:671-675.
10.蔡刚,李闻捷,沈茜.实时逆转录聚合酶链反应绝对定量实验优化的研究.上海医学检验杂志,2003,18:343-346.
11.Rasmussen R.Quantification on the light cycler.Rapid cycle real-time PCR,methods and applications.Heidelberg:Springer Press,2001:21.
12.蔡文琴,王伯云.实用免疫细胞化学与核酸分子杂交技术.成都:四川科学技术出版社,1994:72-89.
13. Weidner N. Intratumor microvessel density as a prognostic factor in cancer. Am J Pathol,1995,147:9-19.
14. Ohno M,Nakamura T , Kunimoto Y, et al . Lymphagenesis correlates with expression of vascular endothelial growth factor-C in Colorectal cancer. Oncol Rep, 2003,10:939-943.
15. Mitas M, Hoover L, Silvestri G, et al. Lunx is a superior molecular marker for detection of non-small cell lung cancer in peripheral blood. J Mol Diagn, 2003, 5: 237-242.
16. Gure AO , Chua R , Williamson B , et al. Cancer-testis genes are coordinately expressed and are markers of poor outcome in non-small cell lung cancer . Clin Cancer Res, 2005, 11: 8055-8062.
17. Cheung ST, Leung KL, Ip YC, et al. Claudin210 expression level is associated with recurrence of primary hepatocellular carcinoma. Clin Cancer Res, 2005, 11: 551-556.
18. Poon RT, Chung KK, Cheung ST, et al. Clinical significance of thrombospondin 1 expression in hepatocellular carcinoma. Clin Cancer Res, 2004, 10: 4150-4157.
19. Sonoda H, Yamamoto K, Kushima R, et al. Detection of lymphnode micrometastasis in gastric cancer by MUC2 RT-PCR: usefulness in pT1 cases. J Surg Oncol, 2004, 88: 63-70.
20. Shimomura K, Sakakura C, Takemura M, et al. Combination of L-3-phosphoserine phosphatase and CEA using real-time RT-PCR improves accuracy in detection of peritoneal micrometastasis of gastric cancer. Anticancer Res, 2004, 24: 1113-1120.
21. Baker MK, Mikhitarian K, Osta W, et al. Molecular detection of breast cancer cells in the peripheral blood of advanced stage breast cancer patients using multimarker real-time reverse transcription-poly-merase chain reaction and a novel porous barrier density gradient centrifugation technology. Clin Cancer Res, 2003, 9: 4865-4871.
22. Masuda TA , Kataoka A , Ohno S, et al. Detection of occult cancer cells in peripheral blood and bone marrow by quantitative RT-PCR assay for cytokeratin-7 in breast cancer patients. Int J Oncol, 2005, 26: 721-730.
23. Patel K, Whelan PJ, Prescott S, et al. The use of real-time reverse transcription-PCR for prostate-specific antigen mRNA to discriminate between blood samples from healthy volunteers and from patients wit h metastatic prostate cancer. Clin Cancer Res, 2004,10:7511-7519.
24. Hara N, Kasahara T, Kawasaki T, et al. Reverse transcription-polymerase chain reaction detection of prostate-specific antigen, prostate-specific membrane antigen, and prostate stem cell antigen in one milliliter of peripheral blood: value for the staging of prostate cancer. Clin Cancer Res, 2002, 8:1794-1799.
25. Buskens CJ , Van Rees BP , Sivula A , et al. Prognostic significance of elevated cyclooxygenase 2 expression in patients with adenocarcinoma of the esophagus. Gastroenterology, 2002, 122: 1800-1807.
26. Sakakibara T, Hibi K, Kodera Y, et al. Plasminogen activator inhibitor-1 as a potential marker for the malignancy of esophageal squamous cell carcinoma. Clin Cancer Res, 2004, 10: 1375 - 1378.
27. Ribuffo D, Gradilone A, Vonella M, et al. Prognostic significance of reverse transcriptase-polymerase chain reaction negative sentinel nodes in malignant melanoma. Ann Surg Oncol, 2003,10: 396-402.
28. Scala S, Ottaiano A, Ascierto PA, et al. Expression of CXCR4 predicts poor prognosis in patients with malignant melanoma. Clin Cancer Res, 2005, 11: 1835-1841.
29. Iinuma H, Okinaga K, Egami H, et al. Usefulness and clinical significance of quantitative real-time RT-PCR to detect isolated tumor cells in the peripheral blood and tumor drainage blood of patients with Colorectal cancer. Int J Oncol, 2006, 28: 297-306.
30. Hernandez BY, Frierson HF, Moskaluk CA, et al. CK20 and CK7 protein expression in Colorectal cancer: demonstration of the utility of a population-based tissue microarray.Hum Pathol, 2005, 36: 275-281.
31. Dandachi N, Balic M, Stanzer S, et al .Critical evaluation of real-time reverse transcriptasepolymerase chain reaction for the quantitative detection of cytokeraitn
20 mRNA incolorectal cancer paitents.J Mol Diagn,2005,7:631-637.
32.苏琪,韩霞,殷红专.CK20诊断大肠癌微转移的价值.世界华人消化杂志,2004,12:2153-2155.
33.徐青,陈瑞新,王志伟,等.大肠癌组织血管生成与患者外周血CK20 mRNA 表达的临床研究.中华医学杂志,2005,85:1205-1208.
34.袁正强,曹建林,陈微微,等.大肠癌患者外周血癌细胞CK20检测及其意义.临床肿瘤学杂志,2005,10:74-76.
35.Berois N,Varangot M,Aizen B,et al.Molecular detection of cancer cells in bone marrow and peripheral blood of patients with operable breast cancer,Comparison of ckl9,Muci and CEA using RT-PCR.European J Cancer,2000,36:717-723.
36.Oberg AN,Lindmark GE,Israelsson AC,et al.Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS.Int J Cancer,2004,111:101-110.
37.Moll R,Schiller DL,Franke WW,et al.Identification of protein IT of the intestinal cytoskeleton as a novel type Ⅰ cytokeratin with unusual properties and expression pattenrs.J Cell Biol,1990,111:567-580.
38.Moll R,Lowe A,Laufer J,et al.Cytokeratin 20 in human carcinomas.A new histodiagnostic marker detected by monoclonal antibodies.Am J Pathol,1992;140:427-447.
39.Aihara T,Fujiwara Y,Miyake Y,et al.Mammaglobin B gene as a novel marker for lymphnode micrometastasisin patients with abdominal cancer.Cancer Lett,2000,150:79-84.
40.陈英剑.荧光实时逆转录PCR定量研究进展.国外医学临床生物化学与检验学分册,2004,25:348-351.
41.Bustin SA.Quantification of mRNA using real-time reverse transcription PCR (RT-PCR):trends and problems.J Mol Endocrinol,2002,29:23-39.
42.陈梓宏,梁荣,赖振南,等.大肠癌患者外周血CEAmRNA和CK20mRNA 的联合检测及临床意义.四川肿瘤防治,2007,20:100-102.
43.易秉强,肖白,吴燕.实时定量RT-PCR检测大肠癌病人外周血CK20mRNA 和CEAmRNA的研究.中华腹部疾病杂志,2005,5:391-394.
44.Zhang XW,Yang HY,Fan P,et al.Detection of micrometastasis in peripheral blood by multi-sampling in patients with colorectal cancer.World J Gastroenterol,2005,11:436-438.
45.Nishizaki T,Matsumata T,Kanematsu T,et al.Surgical manipulation of VX2carcinoma in the rabbit liver evokes enhancement of metastasis.J Surg Res,1990,49:92-97.
46.Garcia-Olmo D,Ontanon J,Garcia-Olmo DC,et al.Experimental evidence does not support use of the "no-touch" isolation technique in colorectal cancer.Dis Colon Rectum,1999,42:1449-1456.
47.Yamaguchi K,Takagi Y,Aoki S,et al.Significant detection of circulating cancer cells in the blood by reverse transcriptase-polymerase chain reaction during colorectal cancer resection.Ann Surg,2000,232:58-65.
48.Weitz J,Koch M,Kienle P,et al.Detection of hematogenic tumor cell dissemination in patients undergoing resection of liver metastases of colorectal cancer.Ann Surg,2000,232:66-72.
49 Weitz J,Kienle P,Magener A,et al.Detection of disseminated colorectal cancer cells in lymph nodes,blood and bone marrow.Clin Cancer Res,1999,5:1830-1836.
50.林国乐,邱辉忠,徐彤,等.大肠癌患者术前和术中外周血微转移的检测及其临床意义.中华普通外科杂志,2002:17:605-607.
51.Dalal S,Berry AM,Cullinane CJ,et al.Vascular endothelial growth factor:a therapeutic target for tumors of the Ewing's sarcoma family.Clin Cancer Res,2005,11:2364-2378.
52.Boxer GM,Tsiompanou E,Levine T,et al.Immunohistochemical expression of vascular endothelial growth factor and microvessel counting as prognostic indicators in node-negative colorectal cancer.Tumour Biol,2005,26:1-8.
53.Uzzan B,Nicolas P,Cucherat M,et al.Microvessel density as a prognostic factor in women wit h breast cancer:a systematic review of the literature and meta-analysis.Cancer Res,2004,64:2941-2955.
54.Giatromanolaki A.Prognostic role of angiogenesis in non-small cell lung cancer.Anticancer Res,2001,21:4373-4382.
55.Sonmezer M,Gungor M,Ensari A,et al.Prognostic significance of tumor angiogenesis in epithelial ovarian cancer:in association with transforming growth factor beta and vascular endothelial growth factor.Int J Gynecol Cancer,2004,14:82-88.
56.Witte MH,Bernas M J,Martin CP,et al.Lymphangiogenesis and lymphangiodysplasia:from molecular to clinical lymphology.Microsc Res Tech,2001,55:122-145.
57.Niakosari F,Kahn HJ,Marks A,et al.Detection of lymphatic invasion in primary melanoma with monoclonal antibody D2-40:a new selective immunohistochemical marker of lymphatic endothelium.Arch Dermatol,2005,141:440-444.
58.Shields JD,Borsetti M,Rigby H,et al.Lymphatic density and metastatic spread in human malignant melanoma.Br J Cancer,2004,90:693-700.
59.牟江洪,阎晓初,李增鹏,等.结直肠癌淋巴管生成的特点及其临床病理意义.中华病理学杂志,2005,34:348-352.
60.阎晓初,牟江红,柳凤轩.大肠癌组织淋巴管分布特点及其与转移和预后的关系.第三军医大学学报,2005,27:2255-2258.
61.姜金波,朱民,李雪梅,等.结肠癌VEGF-C表达、淋巴管密度与淋巴转移和预后的关系.山东大学学报(医学版),2005,43:616-620.
1.周晓东,吕农华.大肠癌的流行病学研究现状.现代消化及介入诊疗,2006,11:149-151.
2.Midgley RS,Kerr DJ.ABC of colorectal cancer:adjuvant therapy.BMJ,2000,321:1208-1211.
3.Habets GG,Scholtes EH,Zuydgeest D.Identification of an invasion-inducing gene,Tiam-1,that encodesa protein with homology to GDP-GTP ecchangers for Rho-like protein.Cell,1994,77:537-549.
4.Mertens AE,Roovers RC,Collard JG.Regulation of Tiaml-Rac signalling.FEBS Lett,2003,546:11-16.
5.Minard ME,Kim LS,Price JE,et al.The role of the guanine nucleotide exchange factor Tiam 1 in cellular migration,invasion,adhesion and tumor progression.Breast Cancer Res Treat,2004,84:21-32.
6.Bourguignon LY,Zhu H,Shao L,et al.Ankyrin-Tiaml interaction promotes Racl signaling and metastatic breast tumor cell invasion and migration.J Cell Biol,2000,150:177-191.
7.Malliri A,van der Kammen RA,Clark K,et al.Mice deficient in the Rac activator Tiam 1 are resistant to Ras-induced skin tumours.Nature,2002,417:867-871.
8.钟大平,周进明,章容,等.Tiam1和Rac1在大肠癌组织中的表达及其临床意义.中华肿瘤防治杂志,2007,14:129-132.
9.Minard ME,Herynk MH,Collard JG,et al.The guanine nucleotide exchange factor Tiaml increases colon carcinoma growth at metastatic sites in an orthotopic nude mouse model.Oncogene,2005,24:2568-2573.
10.Pitti RM,Marsters SA,Lawrence DA,et al.Genomic amplification of a decoy receptor for Fas ligand in lung and colon cancer.Nature,1998,396:699-703.
11.Bai C,Connolly B,Metzker ML,et al.Overexpression of M68/DcR3 in human gastrointestinal tract tumors independent of gene amplification and its location in a four-gene cluster.Proc Natl Acad Sci U S A,2000,97:1230-1235.
12.Yu KY,Kwon B,Ni J,et al.A newly identified member of tumor necrosis factor receptor superfamily(TR6) suppresses LIGHT-mediated apoptosis.J Biol Chem,1999,274:13733-13736.
13.Wu Y,Han B,Sheng H,et al.Clinical significance of detecting elevated serum DcR3/TR6/M68 in malignant tumor patients.Int J Cancer,2003,105:724-732.
14.Takahama Y,Yamada Y,Emoto K,et al.The prognostic significance of overexpression of the decoy receptor for Fas ligand(DcR3) in patients with gastric carcinomas.Gastric Cancer,2002,5:61-68.
15.卢佩颖,张红河.TIAM1与KAI1基因在胃癌组织中的表达研究.浙江临床医学,2005,7:725.
16.黄志为,马忠森.DcR3基因检测在恶性肿瘤研究中的地位.广西医科大学学报,2004,21:182-184.
17.Burchill SA,Bradbury MF,Pittman K,et al.Detection of epithelial cancer cells in peripheral blood by reverse transcriptase polymerase chain reaction.Br J Cancer,1995,71:278-281.
18.蔡刚,李闻捷,沈茜.实时逆转录聚合酶链反应绝对定量实验优化的研究.上海医学检验杂志,2003,18:343-346.
19.Rasmussen R.Quantification on the light cycler.Rapid cycle real-time PCR,methods and applications.Heidelberg:Springer Press,2001:21.
20.Zheng Y.Db1 family guanine nucleotide exchange factors.Trends Biochem Sci,2001,26:724-732.
21.Robbe K,Otto-Bruc A,Chardin P,et al.Dissociation of GDP dissociation inhibitor and membrane translocation are required for efficient activation of Rac by the Db1 homology region of Tiam.J Biol Chem,2003,278:4756-4762.
22.Crompton AM,Foley LH,Wood A,et al.Regulation of Tiaml nucleotide exchange activity by pleckstrin domain binding ligands.J Biol Chem,2000,275:25751-25759.
23.Stam JC,Sander EE,Michiels F,et al.Targeting of Tiaml to the plasma membrane requires the cooperative function of the N-terminal pleckstrin homology domain and an adjacent protein interaction domain.J Biol Chem,1997,272:28447 -28454.
24.李新元,张建中,王登顺,等.肿瘤侵袭诱导基因蛋白在正常及癌组织中的表达及意义.中华病理学杂志,2000,29:294-295
25.Malliri A,Colard JG.Role of Rho-family proteins in cell adhesion and cancer.Curr Opin Cell Biol,2003,15:583-589.
26.Schmidt A,Hall A.Guanine nucleotide exchange factors for Rho GTPases:turning on the switch.Genes Dev,2002,16:1587-1609.
27.Fleming IN,Elilott CM,Buchanan FG,et al.Ca~(2+)/calmodulin-dependent protein kinase Ⅱ regulates Tiam 1 by reversible portein phosphorylation.J Biol Chem,1999,274:12753-12758.
28.Lambert JM,Lambert QT,Reuther GW,et al.Tiaml mediates Ras activation of Rac by a PI(3) K-independent mechanism.Nat Cell Biol,2002,4:621-625.
29.Buchsbaum RJ,ConnollyBA,FeigLA.Interaction of Rac exchange factors Tiaml and Ras-GRF1 with a scaffold for the p38 mitogen-activated protein kinase cascade.Mol Cell Biol,2002,22:4073-4085.
30.Matsuo N,Terao M,Nabeshima Y,et al.Roles of STEF/Tiaml,guanine nucleotide exchange factors for Racl,in regulation of growth cone morphology.Mol Cell Neurosci,2003,24:69-81.
31.Kawauchi T,Chihama K,Nabeshima Y,et al.The in vivo roles of STEF/Tiaml,Racl and JNK in cortical neuronal migration.EMBO J,2003,22:4190-4201.
32.Kunda P,Paglini G,Quiroga S,et al.Evidence for the involvement of Tiaml in axon formation.J Neurosci,2001,21:2361-2372.
33.van Leeuwen FN,van der Kammen RA,Habets GG,et al.Oncogenic activity of Tiam 1 and Racl in NIH3T3 cells.Oncogene,1995,11:2215-2221.
34.Bourguignon LY,Zhu H,Shao L,et al.CD44 interaction with tiaml promotes Racl signaling and hyaluronic acid-mediated breast tumor cell migration.J Biol Chem,2000,275:1829-1838.
35.朱金明,余佩武.T淋巴瘤侵袭转移诱导因子1在胃癌组织中的表达及其临床意义.现代肿瘤医学,2005,13:320-323.
36.朱金明,余佩武,赵永亮.Tiam 1与胃癌细胞侵袭转移及与裸鼠移植瘤形成的关系.中国普通外科杂志,2007,16:855-858.
37.吴明富,史艳燕,韩志强,等.人卵巢癌组织中Tiam 1和Rac 1的表达与肿瘤侵袭转移的关系.中国肿瘤临床,2005,32:910-914.
38.莫立根,王会河,黄光武,等.Tiam 1基因表达与鼻咽癌浸润转移的关系.临床耳鼻咽喉科杂志,2005,19:785-787.
39.刘莉,许岸高,杨玉芳,等.Tiam 1对结直肠癌细胞生物学特性的影响.中华病理学杂志,2005,34:664-668.
40.Engers R,Springer E,Michiels F,et al.Rac affects invasion of human renal cell carcinomas by up-regulating tissue inhibitor of metalloproteinases(TIMP)-1 and TIMP-2 expression.J Biol Chem,2001,276:41889-41897.
41.Hordijk PL,ten Klooster JP,van der Kanunen RA,et al.Inhibition of invasion of epithehal cells by Tiaml-Rac signaling.Science,1997,278:1464-1466.
42.Servitja JM,Marinissen MJ,Sodhi A,et al.Rac 1 function is required for Src-induced transformation:evidence of a role for Tiara 1 and Vav2 in Rac aetivation by Src.J Biol Chem,2003,278:34339-34346.
43.Bourguignon LY,Zhu H,Shao L,et al.Ankyrin-Tiaml interaction promotes Racl signaling and metastatic breast tumor cell invasion and migration.J Cell Biol,2000,150:177-191.
44.Otsuld Y,Tanaka M,Kanlo T,et al.Guanine nucleotide exchange factor,Tiam1,directly binds to c-Myc and interferes with c-Myc-mediated apoptosis in rat-1 fibroblasts.J Biol Chem,2003,278:5132-5140.
45.Bourguignon LY.CD44-mediated oncogenic signaling and cytoskeleton activation during mammary tumor progression.J Mammary GIand Biol Neoplasia,2001,6:287-297.
46.Li H,Zhang L,Lou H,et al.Overexpression of decoy receptor 3 in precancerous lesions and adenocarcinoma of the esophagus.Am J Clin Pathol,2005,124:282-287.
47.Chen J,Zhang L,Kim S.Quantification and detection of DcR3,a decoy receptor in TNFR family.J Immunol Methods,2004,285:63-70.
48. Arakawa Y, Tachibana O, Hasegawa M, et al. Frequent gene amplification and overexpression of decoy receptor 3 in glioblastoma. Acta Neuropathol, 2005, 109: 294-298.
49. Ohshima K, Haraoka S, Sugihara M, et al. Amplification and expression of a decoy receptor for fas ligand (DcR3) in virus (EBV or HTLV-I) associated lymphomas. Cancer Lett, 2000, 160: 89-97.
50. Tsuji S, Hosotani R, Yonehara S, et al. Endogenous decoy receptor 3 blocks the growth inhibition signals mediated by Fas ligand in human pancreatic adenocarcinoma. Int J Cancer, 2003, 106:17-25.
51. Shen HW, Gao SL, Wu YL, et al. Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis. World J Gastroenterol, 2005, 11: 5926-5930.
52. Maeda T, Hao C, Tron VA. Ultraviolet light (UV) regulation of the TNF family decoy receptors DcR2 and DcR3 in human keratinocytes. J Cutan Med Surg, 2001, 5: 294-298.
53. Mild G, Bachmann F, Boulay JL, et al. DCR3 locus is a predictive marker for 5-fluorouracil-based adjuvant chemotherapy in Colorectal cancer. Int J Cancer, 2002, 102: 254-257.
1. Clements J, Hooper J, Dong Y, et al. The expanded human kallikrein (KLK) gene family: genomic organisation, tissue-specific expression and potential functions. Biol Chem, 2001, 382: 5-14.
2. Diamandis EP, Yousef GM, Clements J, et al. New nomenclature for the human tissue kallikrein gene family. Clin Chem, 2000,46: 1855-1858.
3. Yousef GM, Obiezu CV, Luo LY, et al. Human tissue kallikreins: from gene structure to function and clinical applications. Adv Clin Chem, 2005,39:11-79.
4. Yousef GM, Diamandis EP. The new human tissue kallikrein gene family: structure, function, and association to disease. Endocr Rev, 2001,22: 184-204.
5. Borgono CA, Diamandis EP. The emerging roles of human tissue kallikreins in cancer. Nat Rev Cancer, 2004,4: 876-890.
6. Borgono CA, Michael IP, Diamandis EP. Human tissue kallikreins: physiologic roles and applications in cancer. Mol Cancer Res, 2004,2: 257-280.
7. Diamandis EP. Prostate-specific antigen: its usefulness in clinical medicine. Trends Endocrinol Metab, 1998, 9: 310-316.
8. Anisowicz A, Sotiropoulou G, Stenman G, et al. A novel protease homolog differentially expressed in breast and ovarian cancer. Mol Med, 1996, 2: 624-636.
9.Yamashiro K, Tsuruoka N, Kodama S, et al. Molecular cloning of a novel trypsin-like serine protease (neurosin) preferentially expressed in brain. Biochim BiophysActa, 1997,1350: 11-14.
10. Little SP, Dixon EP, Norris F, et al. Zyme, a novel and potentially amyloidogenic enzyme cDNA isolated from Alzheimer's disease brain. J Biol Chem, 1997, 272: 25135-25142.
11. Diamandis EP, Scorilas A, Fracchioli S, et al. Human kallikrein 6 (hK6): a new potential serum biomarker for diagnosis and prognosis of ovarian carcinoma. J Clin Oncol, 2003, 21: 1035-1043.
12. Hoffman BR, Katsaros D, Scorilas A, et al. Immunofluorometric quantitation and histochemical localisation of kallikrein 6 protein in ovarian cancer tissue:a new independent unfavourable prognostic biomarker.Br J Cancer,2002,87:763-771.
13.Tanimoto H,Underwood LJ,Shigemasa K,et al.Increased expression of protease M in ovarian tumors.Tumour Biol,2001,22:11-18.
14.Wolf WC,Evans DM,Chao L,et al.A synthetic tissue kallikrein inhibitor suppresses cancer cell invasiveness.Am J Pathol,2001,159:1797-1805.
15.Diamandis EP,Yousef GM,Soosaipillai AR,et al.Immunofluorometric assay of human kallikrein 6(zyme/protease M/neurosin) and preliminary clinical applications.Clin Biochem,2000,33:369-375.
16.Yousef GM,Borgono CA,Popalis C,et al.In-silico analysis of kallikrein gene expression in pancreatic and colon cancers.Anticancer Res,2004,24:43-51.
17.Ogawa K,Utsunomiya T,Mimori K,et al.Clinical significance of human kallikrein gene 6 messenger RNA expression in colorectal cancer.Clin Cancer Res,2005,11:2889-2893.
18.Ghosh MC,Grass L,Soosaipillai A,et al.Human kallikrein 6 degrades extracellular matrix proteins and may enhance the metastatic potential of tumour cells.Tumour Biol,2004,25:193-199.
19.Iwata A,Maruyama M,Akagi T,et al.Alpha-synuclein degradation by serine protease neurosin:implication for pathogenesis of synucleinopathies.Hum Mol Genet,2003,12:2625-2635.
20.Burchill SA,Bradbury MF,Pittman K,et al.Detection of epithelial cancer cells in peripheral blood by reverse transcriptase polymerase chain reaction.Br J Cancer,1995,71:278-281.
21.蔡刚,李闻捷,沈茜.实时逆转录聚合酶链反应绝对定量实验优化的研究.上海医学检验杂志,2003,18:343-346.
22.Rasmussen R.Quantification on the light cycler.Rapid cycle real-time PCR,methods and applications.Heidelberg:Springer Press,2001:21.
23.Schmaier AH.The plasma kallikrein-kinin system counterbalances the renin-angiotensin system.J Clin Invest,2002,109:1007-1009.
24.Diamandis EP,Yousef GM,Luo LY,et al.The new human kallikrein gene family:implications in carcinogenesis.Trends Endocrinol Metab,2000,11:54-60.
25.Yu H,Levesque MA,Clark GM,et al.Prognostic value of prostate-specific antigen for women with breast cancer:A large United States cohort study.Clin Cancer Res,1998,4:1489-1497.
26.Yousef GM,Diamandis EP.Human kallilreins:common structural features,sequence analysis and evaluation.Curr Genomics,2003,276:127-232.
27.Clements JA,Willemsen NM,Myers SA,et al.The tissue kallikrein family of serine proteases:functional roles in human disease and potential as clinical biomarkers.Crit Rev Clin Lab Sci,2004,41:265-312.
28.Jin E,Fujiwara M,Pan X,et al.Protease-activated receptor(PAR)-1 and PAR-2participate in the cell growth of alveolar capillary endothelium in primary lung adenocarcinomas.Cancer,2003,97:703-713.
29.Schuster R,Max N,Mann B,et al.Quantitative real-time RT-PCR for detection of disseminated tumor cells in peripheral blood of patients with colorectal cancer using different mRNA markers.Int J Cancer,2004,108:219-227.
30.Lu KH,Patterson AP,Wang L,et al.Selection of potential markers for epithelial ovarian cancer with gene expression arrays and recursive descent partition analysis.Clin Cancer Res,2004,10:3291-3300.
31.Ni X,Zhang W,Huang KC,et al.Characterisation of human kallikrein6/protease M expression in ovarian cancer.Br J Cancer,2004,91:725-731.
32.胡成进,陈英剑,闻新棉,等.KLK6 mRNA在乳腺癌中的表达及其临床病理和生物学意义.解放军医学杂志,2007,32:1085-1087.
33.Petraki CD,Gregorakis AK,Vaslamatzis MM,et al.Prognostic implications of the immunohistochemical expression of human kallikreins 5,6,10 and 11 in renal cell carcinoma.Tumour Biol,2006,27:1-7.
34.Nagahara H,Mimori K,Utsunomiya T,et al.Clinicopathologic and biological significance of kallikrein 6 overexpression in human gastric cancer.Clin Cancer Res,2005,11:6800-6806.
35.Liotta LA,Steeg PS,Stetler-Stevenson WG.Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation.Cell,1991,64:327-336.
36.Tryggvason K,Hoyhtya M,Salo T.Proteolytic degradation of extracellular matrix in tumor invasion.Biochim Biophys Acta,1987,907:191-217.
37.Duffy MJ.The role of proteolytic enzymes in cancer invasion and metastasis.Clin Exp Metastasis,1992,10:145-155.
38.Aznavoorian S,Murphy AN,Stetler-Stevenson WG,et al.Molecular aspects of tumor cell invasion and metastasis.Cancer,1993,71:1368-1383.
39.Matrisian LM.Cancer biology:extracellular proteinases in malignancy.Curr Biol,1999,9:R776-778.
40.Nelson AR,Fingleton B,Rothenberg ML,et al.Matrix metalloproteinases:biologic activity and clinical implications.J Clin Oncol,2000,18:1135-1149.
41.McCawley LJ,Matrisian LM.Tumor progression:defining the soil round the tumor seed.Curr Biol,2001,11:R25-27.
42.Magklara A,Mellati AA,Wasney GA,et al.Characterization of the enzymatic activity of human kallikrein 6:Autoactivation,substrate specificity,and regulation by inhibitors.Biochem Biophys Res Commun,2003,307:948-955.
43.Bernett MJ,Blaber SI,Scarisbrick IA,et al.Crystal structure and biochemical characterization of human kallikrein 6 reveals that a trypsin-like kallikrein is expressed in the central nervous system.J Biol Chem,2002,277:24562-24570.
44.Yousef GM,Luo LY,Scherer SW,et al.Molecular characterization of zyme/protease M/neurosin(PRSS9),a hormonally regulated kallikrein-like serine protease.Genomics,1999,62:251-259.
45.Yousef GM,Diamandis EP.Kallikreins,steroid hormones and ovarian cancer:is there a link? Minerva Endocrinol,2002,27:157-166.
46.张芳,陈英剑,胡成进.17β雌二醇对人卵巢癌细胞株H08910 KLK6基因表达的调节.山东大学学报(医学版),2007,45:42-45.
47.陈英剑,胡成进,闻新棉,等.雌激素对人乳腺癌细胞株MCF-7 KLK6mRNA和蛋白表达水平的影响.中国病理生理杂志,2007,23:246-249.
48.Qiu Y,Langman MJ,Eggo MC.Targets of 17 beta-oestradiol-induced apoptosis in colon cancer cells:a mechanism for the protective effects of hormone replacement therapy? J Endocrinol, 2004, 181: 327-337.
49. Campbell-Thompson M, Lynch IJ, Bhardwaj B. Expression of estrogen receptor (ER) subtypes and ERbeta isoforms in colon cancer. Cancer Res, 2001,61: 632-640.
50. Konstantinopoulos PA, Kominea A, Vandoros G, et al. Oestrogen receptor beta (ERbeta) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumour's dedifferentiation. Eur J Cancer, 2003, 39: 1251-1258.