半边莲煎剂对肝癌H22荷瘤小鼠的抑瘤作用及对P27、Bcl-2及Survivin表达的影响
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摘要
目的：以肝癌H22荷瘤小鼠为研究对象,通过对比应用半边莲煎剂、5-氟脲嘧啶(5-FU)、生理盐水后移植瘤生长转移的情况,观察半边莲煎剂的疗效；并利用免疫组化的方法,检测P27、Bcl-2及Survivin表达情况,研究和讨论半边莲煎剂抑制肝癌的作用机制。
方法：建立肝癌H22荷瘤小鼠移植瘤模型,采用随机法将24只小鼠分为三组,半边莲煎组、5-氟脲嘧啶组、生理盐水组各8只。通过测定肝癌小鼠的瘤重、抑瘤率及胸腺、脾腺指数,观察半边莲煎剂对肿瘤生长的影响；采用免疫组化技术、图像分析技术定量检测P27、Bcl-2、Survivin的表达情况,运用统计学分析数据的意义。
结果：1.半边莲煎剂能使肝癌小鼠的瘤重减少,抑瘤率为33.98%,显著低于对照组(P＜0.05)；2.半边莲煎剂可以提高肝癌小鼠的胸腺、脾的重量,增加免疫能力,较对照组、5-FU组对比,有显著意义(P＜0.05);3.P27、Bcl-2、Survivin在三组中均呈阳性表达,单因素方差分析表明：半边莲煎剂组P27表达增强、Bcl-2表达减弱、Survivin表达减弱,与对照组相比均有显著性差异(P＜0.05),半边莲煎剂组与5-FU组比较(P＞0.05),三者均无统计学意义。
结论：半边莲煎剂具有抑制肝癌生长,显著提高肝癌小鼠的免疫功能的作用；半边莲煎剂可能通过提高P27蛋白表达、减少Bcl-2、Survivn蛋白表达,从而起到抗肿瘤的作用,其作用机制可能与P27、Bcl-2和Survivin的表达有关。
Objective:H22 tumor-bearing mice with liver cancer were established for the study, to observe the effcts of Chinese Lobelia Herb decoction (CLH) by comparing the applications of CLH、5 florouracilu chemotherapy treatment (5-Fu) and normal control inhibiting liver cancer growth and metastasis in tumor-bearing mice. Immunohistochemical method was used to detect the expressions ofP27, Bcl-2 and Survivin to study and discuss its tumor-inhibiting mechanism of Chinese Lobelia Herb decoction.
Methods:Established the mice models of transplantation tumor of H22 carcinomatous.With randomized method categorized 24 mice into 3 groups including Chinese Lobelia Herb decoction group,5-fluorouracil chemo-therapy treatment group and normal control group.The inhiting rate of tumor, the thymus index and spleen index were detectioned to observe the effect of CLH on tumor growth. The methods of immunhistochemistry and image pattern analysis were taken to quantitatively detect and compared the expressions of P27、Bcl-2 and Survivin among the three groups. Last analyzed the statistical significance of datas.
Results:1.Chinese Lobelia Herb decoction was shown to reduce tumor weights that the inhibiting rate of tumor was 33.98%with significant difference compared with normal control(P＜0.05); 2. Chinese Lobelia Herb decoction was shown that it could increase the weights of thymus and spleen, moreover it could increase the immunity and improve the quality of life in the mice with significant difference compared with normal control group and 5-FU group (P＜0.05); 3.The expressions of P27、Bcl-2 and Survivin in the three groups were positive. ANOVA analysis manifests that: the expression of P27 was increased meanwhile the expressions of Bcl-2 and Survivin were decreased in Chinese Lobelia Herb decoction group.They were all statistical significances compared with normal control group (P＜0.05).The comparison between CLH group and 5-Fu group were not statistically different (P＞0.05)
Conclusion:Chinese Lobelia Herb decoction could inhibit the growth of liver cancer, significantly increase the immunity and improve the quality of life in the mice; Chinese Lobelia Herb decoction was also shown significant anti-tumor effect through the expression of P27 increased and the expressions of Bcl-2、Survivn reduced.The mechanism may be relative to the expressions of P27、Bcl-2 and Survivin.
方法：建立肝癌H22荷瘤小鼠移植瘤模型,采用随机法将24只小鼠分为三组,半边莲煎组、5-氟脲嘧啶组、生理盐水组各8只。通过测定肝癌小鼠的瘤重、抑瘤率及胸腺、脾腺指数,观察半边莲煎剂对肿瘤生长的影响；采用免疫组化技术、图像分析技术定量检测P27、Bcl-2、Survivin的表达情况,运用统计学分析数据的意义。
结果：1.半边莲煎剂能使肝癌小鼠的瘤重减少,抑瘤率为33.98%,显著低于对照组(P＜0.05)；2.半边莲煎剂可以提高肝癌小鼠的胸腺、脾的重量,增加免疫能力,较对照组、5-FU组对比,有显著意义(P＜0.05);3.P27、Bcl-2、Survivin在三组中均呈阳性表达,单因素方差分析表明：半边莲煎剂组P27表达增强、Bcl-2表达减弱、Survivin表达减弱,与对照组相比均有显著性差异(P＜0.05),半边莲煎剂组与5-FU组比较(P＞0.05),三者均无统计学意义。
结论：半边莲煎剂具有抑制肝癌生长,显著提高肝癌小鼠的免疫功能的作用；半边莲煎剂可能通过提高P27蛋白表达、减少Bcl-2、Survivn蛋白表达,从而起到抗肿瘤的作用,其作用机制可能与P27、Bcl-2和Survivin的表达有关。
Objective:H22 tumor-bearing mice with liver cancer were established for the study, to observe the effcts of Chinese Lobelia Herb decoction (CLH) by comparing the applications of CLH、5 florouracilu chemotherapy treatment (5-Fu) and normal control inhibiting liver cancer growth and metastasis in tumor-bearing mice. Immunohistochemical method was used to detect the expressions ofP27, Bcl-2 and Survivin to study and discuss its tumor-inhibiting mechanism of Chinese Lobelia Herb decoction.
Methods:Established the mice models of transplantation tumor of H22 carcinomatous.With randomized method categorized 24 mice into 3 groups including Chinese Lobelia Herb decoction group,5-fluorouracil chemo-therapy treatment group and normal control group.The inhiting rate of tumor, the thymus index and spleen index were detectioned to observe the effect of CLH on tumor growth. The methods of immunhistochemistry and image pattern analysis were taken to quantitatively detect and compared the expressions of P27、Bcl-2 and Survivin among the three groups. Last analyzed the statistical significance of datas.
Results:1.Chinese Lobelia Herb decoction was shown to reduce tumor weights that the inhibiting rate of tumor was 33.98%with significant difference compared with normal control(P＜0.05); 2. Chinese Lobelia Herb decoction was shown that it could increase the weights of thymus and spleen, moreover it could increase the immunity and improve the quality of life in the mice with significant difference compared with normal control group and 5-FU group (P＜0.05); 3.The expressions of P27、Bcl-2 and Survivin in the three groups were positive. ANOVA analysis manifests that: the expression of P27 was increased meanwhile the expressions of Bcl-2 and Survivin were decreased in Chinese Lobelia Herb decoction group.They were all statistical significances compared with normal control group (P＜0.05).The comparison between CLH group and 5-Fu group were not statistically different (P＞0.05)
Conclusion:Chinese Lobelia Herb decoction could inhibit the growth of liver cancer, significantly increase the immunity and improve the quality of life in the mice; Chinese Lobelia Herb decoction was also shown significant anti-tumor effect through the expression of P27 increased and the expressions of Bcl-2、Survivn reduced.The mechanism may be relative to the expressions of P27、Bcl-2 and Survivin.
引文
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[10]Hsu YL,Kuo PL,Tzeng WS,et al.Chalcone inhibits the proliferation of human breast cancer cell by blocking cell cycle progression and inducing apoptosis.Food Chem Toxicol,2006,44:704-713.
[11]彭民浩,韦建宝,陈滨等.P27在肝细胞癌中的表达及其临床意义.2004,10(1)：48-51
[12]Tannapfel A,Grund D,Katalinic A,et al.Decreased expression of p27 protein is associated with advanced tumors stage in hepatocellular carcinomaInt.J Cancer,
2000,89(4):350-355
[13]王科,王学浩,张峰.P27在肝细胞肝癌组织中的表达及意义.肝胆外科杂志,2002,10(1)：71-72
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[15]Fiorention M,Altimari A,et al.Acquired Expression of p27 Is a Favorable Prognostic Indicator in Patients with Hepatocellular Carcinoma.Clin Cancer Res,2000,6(10):3966-3972
[16]Tsujimoto Y,Cossman J,Jafe,et al.Involvement ofbel-2 gene in human follocular lymphoma[J].Science,1985,228(4706):1440-1443
[17]Mathai JP,Germain M,Shore GC.BH3-only BIK regulates BAXBAK-dependent release of Ga2+from endoplasmic reticulum stores and mitochondrial apoptosis during stress-induced cell death[J].J Biol Chem,2005,280:23829-23836
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[20]Brooks C,Dong Z.Regulation of mitochondrial morphological dynamics during poptosis by Bcl-2 family proteins:a key in Bak[J].Cell Cycle,2007,6(24): 3043-3047
[21]Adams JM,Cotry S.The Bcl-2 protein family:arbiters of cell survival [J]. Science,1998,281(5381):1322-1326
[22]Takahashi M,Saito H,Okuyama T,et al.Overexpression of bcl-2 proects human hepatoma cells from Fas-antibody-mediated apoptosis[J].J Hepatol,1999,31,(2): 315-322
[23]Zeppa P,Benincasa G,Fulciniti F,et al.Apoptosis and cytologic differentiation in hepatocullular carcinima on fine needle aspiration samples.Acta Cytol,1996, 40(5):861
[24]王万忠,王卫屏,王家耀等.Bcl-2蛋白在肝肿瘤组织的表达及意义.中华肝脏病杂志,1996,4(4)：207
[25]Beerheide W,Tan YJ,Teng E,et al.Downregulation of proapoptotic proteins bax and bcl-X(S) in p53 overexpressing hepaocellular carcinomas[J].Biochem
Biophys Res Commun,2000.273(1):54-61
[26]刘战培,严律南.Bcl-2蛋白在肝细胞肝癌发生过程中的表达及其与肝细胞凋亡变化的关系.中国普外基础与临床杂志,1999,6(4)：193-195
[27]Ambrosini G,Adida C,Altieri DC.Anove 1 anti-apoptgene survivin expressed in cancer and lymphoma [J].Nature Med,1997,3(8):917-921
[28]陈明,伊新,林木生等.Survivin在肝癌中的表达及与P53表达的关系[J].中华肝胆外科杂志,2004,10(5)：332
[29]Suzuki A,Ito T,Kawano H,et al.Survivin initiates pro-caspase 3/p21 complex formation as a result of interaction with Cdk4 to resist Fas-mediated cell death[J].Onco-gene,2000,19(10):1346-1353
[30]Papapetropoulos A,Fulton D,Mahboubi K,et al.Angiopoietin-1 inhibits En-dothelial cell apoptosis via the Akt/survivin pathway[J].J Biol Chem,2000,275 (13):9102-9105
[31]Ito T,Shiraki K,Sugimoto K,et al.Survivin promotes cell proliferation in human hepatocellular carcinoma [J].Hepatology,2000,31(5):1080-1085
[32]王颖,王家龙.Survivin在原发性肝细胞癌中的表达及意义[J].中华消化杂志,2002,22(8)：463-466
[33]Ikeguchi M,Ueta T,Yamane Y,et al.Inducible nitric oxide synthase and survivin messenger RNA expression in hepatocellular carcinoma.Clin Cancer Res,2002, 8(10):3131-3136
[34]Satob K,Kaneko K,Hirota M.Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors.Cancer,2001,92(2):271-278
[35]王颖,王家龙.生存素反义寡核苷酸诱导肝癌细胞凋亡的实验研究[J].中华消化杂志,2003,23(1)：11-14
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