靶向Rb的siRNA真核表达载体与纳米磁小体靶向药囊联合作用人胆管癌的实验研究
|
摘要
研究目的
胆管癌主要指源于肝外胆管的恶性肿瘤,预后不佳。流行病学少见,国外尸检发现率为0.01%～0.5%,占恶性肿瘤总数3%,我国尸检占0.07%～0.3%,其检出率逐年增高。胆管癌占所有胆道疾病总数的2.32%,男女之比为1.46:1。近年来其发病率有上升的趋势。胆管癌的治疗方法多采用以手术为主放化疗为辅的综合胆管癌的治疗方法多采用以手术为主放化疗为辅的综合治疗。但由于其位置深、隐蔽、早期发现困难,使手术治疗效果很差,生存率低。化疗药物通过作用在细胞繁殖的不同环节,抑制或杀灭肿瘤细胞,从而达到治疗的目的,已成为当今临床治疗肿瘤的重要手段。但是目前中晚期胆管癌的化疗效果很不令人满意。既往报道以5-氟尿嘧啶为主的单药或联合化疗的有效率为14%～18%。
运用纳米技术和化疗药物相结合的方法,在磁导向的作用下靶向性治疗恶性肿瘤是近年来兴起的一种新的治疗手段。载药的磁性纳米微粒是纳米技术和化疗药物相结合的产物。在磁导向作用下聚集在肿瘤部位,这不仅提高了肿瘤局部的化疗药物浓度,而且纳米颗粒本身的可控性降解可使载体内的化疗药物持续性释放,使药物在血液中循环时间延长并维持有效的血药浓度,且可减少代谢产物,易于清除,副作用少。为恶性肿瘤的综合治疗带来了新的希望和途径。
在邹声泉教授的领导下,我们课题组不仅成功获得了国家高新技术研究发展计划(863计划)资助的重大课题《纳米生物磁小体靶向药囊治疗肝胆胰恶性肿瘤的实验研究》(编号2002AA214061)。该项目的核心内容是:采用高新技术研制磁化纳米金属薄膜包覆的支架,在通畅胆道引流的同时,可增强5-Fu纳米磁小体靶向药囊的靶向性,真正达到高选择性区域性化疗的目的,以实现胆管癌治疗观念中的创新和突破。而且在前期的研究中我们对胆管癌肿瘤生物学特性有了深入的研究,认识到胆管癌的发生、发展过程中各种致癌因子和抑癌因子中存在着复杂的网络式的反馈机制,其中Rb的蛋白信号转导通路可能在这个网络中起到重要的作用。在肝外胆管癌的分子细胞遗传学研究取得了很大的进展。
在此基础上,我们拟构建针针对Rb的RNAi真核表达载体,并将其转导人胆管癌细胞中,观察对Rb蛋白信号转导通路的影响及对胆管癌生长的影响。并与纳米磁小体靶向药囊联合作用人胆管癌细胞,观察在体内外实验中它们的联合作用对胆管癌细胞生长的影响。
研究方法
根据Rb基因cDNA序列,设计出针对目的基因的4个siRNA靶序列。克隆到真核表达载体pGenesil vector中,通过酶切鉴定和DNA测序鉴定正确与否。将构建好的4个siRNA表达载体分别转染到QBC939细胞中MTT法、FCM,检测Rb基因沉默后胆管癌细胞生长变化、细胞周期和细胞凋亡情况。RT-PCR和Western Blot检测Rb的mRNA和蛋白在胆管癌细胞系中的表达水平,同时检查与Rb相关的基因的表达情况。将QBC939细胞接种于裸鼠皮下,建立胆管癌裸鼠种植模型,观察体内种植瘤的发生率和生长情况。通过急性毒性试验找出TM5-FuNC对裸鼠的半数致死量(LD50)。根据急性毒性试验结果设定合适的药物剂量。MTT检测TM5-FuNC对转染psiRNARb-1前后QBC939细胞生长抑制的影响。将裸鼠模型再分成4组,分别为对照组,5-Fu组, TM5-FuNC组,psiRNARb-1和TM5-FuNC联合组。观察肿瘤体积变化,计算抑瘤率,绘制生长曲线。
实验结果
成功构建了针对Rb的真核表达载体,转染psiRNARb-1质粒48至72小时后QBC939细胞的生长相对于对照组细胞受到了一定程度的抑制,MTT实验结果显示当Rb基因在QBC939细胞中表达沉默时,对细胞生长有一定的抑制作用,在72小时内呈一定的时间依赖性。转染后QBC939细胞中Rb的mRNA表达相较于未转染细胞明显下调,Rb蛋白表达也明显受抑制。p16的mRNA和蛋白的表达变化情况与Rb相反。E2F1转录因子mRNA和蛋白的表达变化趋势与Rb相同。转染组中P27kip1蛋白的表达较未转染组有所增强。TM5-FuNC的急性动物实验中所有动物均未死亡,最大耐受剂量为1250 mg/kg。是临床给药剂量的5倍。TM5-FuNC 250和200mg/kg组的移植肿瘤体积在治疗后35天与对照组比较有显著差异(P<0.05)。电境结果显示,TM5-FuNC对荷瘤裸鼠的肿瘤细胞有诱导凋亡作用,且随着药物浓度的增大,细胞凋亡的形态改变越明显。转染psiRNARb-1的QBC939细胞可显著增强TM5-FuNC和5-Fu的生长抑制作用(P<0.05)。TM5-FuNC和TM5-FuNC+psiRNARb-1联合组与对照组和5-Fu组比较肿瘤体积的生长有明显差异(P<0.001)。TM5-FuNC和TM5-FuNC+psiRNARb-1联合组之间比较(P=0.44)有一定的差异显著性。
实验结论
1成功构建了4对针对Rb的siRNA真核表达载体,其中转染的psiRNARb-1质粒对胆管癌QBC939细胞有一定的抑制作用,并可诱导的胆管癌细胞G0/G1期阻滞和细胞凋亡。这种作用可能跟Rb与p16、E2F1、P27kip1之间的反馈调节机制相关。为我们将Rb作为基因治疗的靶目标提供了实验依据。
2通过急性毒性实验,我们获得了TM5-FuNC最大耐受剂量为后续实验提供了用药依据。
3 TM5-FuNC新剂型在内磁场的导向作用下能有效抑制人胆管癌裸鼠移植瘤的生长,使我们所研制的TM5-FuNC纳米药物制剂结合内磁场靶向治疗胆管癌的设计理念获得了实验的证实即可磁化的NiTi合金支架在保持胆道通畅引流的情况下,能高靶向的引导TM5-FuNC聚集在肿瘤部位实现对肿瘤高效地、持续地的治疗,并保证对机体的低毒性。
4 TM5-FuNC联合psiRNARb-1作用组相比单用TM5-FuNC组能更有效的抑制胆管癌肿瘤的生长,为TM5-FuNC联合基因治疗提供了理论依据和实验平台,为治疗中晚期胆管癌指明了一条新的途径。
Background & Objective: Cholangiocarcinoma is mainly named as the extrahepatic bile duct malignant tumor. The prognosis of it isn’t idealization. It’s scarce in epidemiology and is about 0.01%~0.5% in biopsy, about 3% in the total count of the malignant tumor. It’s about 0.07%~0.3% in domestic biopsy. The incidence rate of Cholangiocarcinoma is about 2.32% in all disease of biliary tract. The rate of female and male is 1.46:1. The incident rate is increasing progressively in recent years. The treatments of cholangiocarcinoma were given priority to operation or combine chemotherapy with radiation treatment. But the position of cholangiocarcinoma very deep and concealment in anatomy, it’s hard to detect cholangiocarcinoma in earlier period. As a result the effect of operation was discontentment, and the survival rate was very low. The chemiotherapy drug inhibit or kill tumor cells by act on different links of cell breeding, which can treat tumor. This is become a important method for clinical tumor treatment now. But the effect of chemiotherapy drug can not reach the aim of satisfactory. The report of the effect of 5-Fu is only reach 14%-18% regardless of simplex or combine medication.
Targeting treatment to malignant tumor is a new therapeutic method which as a result of conjunct nanotechology with chemotherapeutic drug rising in recent years. The principle is that the carrier including various kinds species group affinity carry particles and magnetic targeting particles take drug which could targeted reach to the locus of medication in order to treat disease. This will afford experctation for cure malignant tumor.
Our topic group lead by professor zoushengquan succeed get a great topic supported by the grant from The National Hi-Tech Research and Development Program of China (863 Program)(No. 2002AA214061). The core content of this project is that a magnetize cradle which be sacked by metal can reinforcement the TM5-FuNC targeting power while the cradle keeping bile duct drain smoothly. As a result that this will to achieve the real high selective and regional treatment. This will realizated break and new idea of Cholangiocarci- noma treatment, Furthermore, our group of the topic group has investigated deeply on the cholangiocarcinoma bionomics before. We concluded that there are complex network feed back mechanism between the oncogene and the anti-oncogene which is exist in the process of occurrence and develop of cholangiocarcinoma. pRb signal pathway were close importa- nt to the complex network feedback system of cholangiocarcinoma. It’s a great break in the research of the molecular cytogenetics in the cancer of extrahepatic biliary duct.
We based on the prophase research, put our emphasis on the pRb signal transduction pathway, and make some investigate on the apoptosis of the cholangiocarcinoma and the Fuction of the sensitization of the chemotherapy. We association the siRNA eukaryotic expression vector Rb with TM5-FuNC act on human cholangiocarcinoma cell in vivo and vitro, which will provid us a test base and realization pathway for the union opoerate of nanotechnology and gene therapy. Restricted to the time, we are limited and superficialis level, and we will make the Further step in the Future.
1. Methods and results: Eukaryotic expression vector of siRNA Rb were constructed, and used to transiently transfect to QBC939 mediated with LipofectamineTM 2000. The growth of QBC939 cell has been inhibited in some degree compare to the control group form 48h to 72h after transfection. Rb slience caused cell cycle arrest at G0/G1. Apoptotic cell index also increased in some degree. It show certain time dependency in 72h.Rb mRNA and protein under expression significantly compare to the control after transfection.p16 mRNA and protein expression inversed to Rb. E2F1 have identical change tendency with Rb. No animal dead in Acute toxicity test. The most tolerance dosage of TM5-FuNC is 1250mg/kg, which is 5 times dosage of clinical administration. High and middle dosage groups of TM5-FuNC have difference significantly compare to the control group (P <0.05) at 35 after treatment.The high and middle dosage groups of TM5-FuNC have discovered cells apoptosis under electron microscope. The ratio of growth inhibiting of TM5-FuNC and 5-Fu increased significantly after transfection. The tumor volume of TM5-FuNC group and psiRNARb-1+TM5-FuNC group have greatly difference compare with those of the control group and 5-Fu group (P<0.001). The tumor volume of TM5-FuNC group compare to tumor volume of TM5-FuNC group also have some difference (P=0.44).
Conclusions:
1 We are succeed constructed the eukaryotic expression vector of siRNA Rb which can specifically inhibit Rb expression There are certain inhibited growth effects of QBC939 cell line after transfecting psiRNARb-1 into cell. And Rb slience caused cell cycle arrest at G0/G1. Apoptotic cell index also increased in some degree, which may correlation with the Feedback adjust mechanism of Rb、p16、E2F1、P27kip1 protein . It can be used for later test.
2 We get TM5-FuNC the maximum tolerance dosage by acute toxicity experiment which is provided base for medication.
3 The TM5-FuNC can inhibit the tumor growth of human cholangiocarcinoma xeno- graft on nude mice. It show that our TM5-FuNC and design ideal of infra-magnetic field targeting get confirmation by test. The Ni-Ti alloy cradle who can be magnetized can high targeting led TM5-FuNC collecting in tumor tissue for treatment effectually and lastly when it keeping bile duct drain smoothly.
4 TM5-FuNC + psiRNARb-1 have better inhibit effect for cholangiocarcinoma compare to simplex TM5-FuNC. It would establish a test background for Further inosculatio TM5-FuNC with gene therapy.
胆管癌主要指源于肝外胆管的恶性肿瘤,预后不佳。流行病学少见,国外尸检发现率为0.01%～0.5%,占恶性肿瘤总数3%,我国尸检占0.07%～0.3%,其检出率逐年增高。胆管癌占所有胆道疾病总数的2.32%,男女之比为1.46:1。近年来其发病率有上升的趋势。胆管癌的治疗方法多采用以手术为主放化疗为辅的综合胆管癌的治疗方法多采用以手术为主放化疗为辅的综合治疗。但由于其位置深、隐蔽、早期发现困难,使手术治疗效果很差,生存率低。化疗药物通过作用在细胞繁殖的不同环节,抑制或杀灭肿瘤细胞,从而达到治疗的目的,已成为当今临床治疗肿瘤的重要手段。但是目前中晚期胆管癌的化疗效果很不令人满意。既往报道以5-氟尿嘧啶为主的单药或联合化疗的有效率为14%～18%。
运用纳米技术和化疗药物相结合的方法,在磁导向的作用下靶向性治疗恶性肿瘤是近年来兴起的一种新的治疗手段。载药的磁性纳米微粒是纳米技术和化疗药物相结合的产物。在磁导向作用下聚集在肿瘤部位,这不仅提高了肿瘤局部的化疗药物浓度,而且纳米颗粒本身的可控性降解可使载体内的化疗药物持续性释放,使药物在血液中循环时间延长并维持有效的血药浓度,且可减少代谢产物,易于清除,副作用少。为恶性肿瘤的综合治疗带来了新的希望和途径。
在邹声泉教授的领导下,我们课题组不仅成功获得了国家高新技术研究发展计划(863计划)资助的重大课题《纳米生物磁小体靶向药囊治疗肝胆胰恶性肿瘤的实验研究》(编号2002AA214061)。该项目的核心内容是:采用高新技术研制磁化纳米金属薄膜包覆的支架,在通畅胆道引流的同时,可增强5-Fu纳米磁小体靶向药囊的靶向性,真正达到高选择性区域性化疗的目的,以实现胆管癌治疗观念中的创新和突破。而且在前期的研究中我们对胆管癌肿瘤生物学特性有了深入的研究,认识到胆管癌的发生、发展过程中各种致癌因子和抑癌因子中存在着复杂的网络式的反馈机制,其中Rb的蛋白信号转导通路可能在这个网络中起到重要的作用。在肝外胆管癌的分子细胞遗传学研究取得了很大的进展。
在此基础上,我们拟构建针针对Rb的RNAi真核表达载体,并将其转导人胆管癌细胞中,观察对Rb蛋白信号转导通路的影响及对胆管癌生长的影响。并与纳米磁小体靶向药囊联合作用人胆管癌细胞,观察在体内外实验中它们的联合作用对胆管癌细胞生长的影响。
研究方法
根据Rb基因cDNA序列,设计出针对目的基因的4个siRNA靶序列。克隆到真核表达载体pGenesil vector中,通过酶切鉴定和DNA测序鉴定正确与否。将构建好的4个siRNA表达载体分别转染到QBC939细胞中MTT法、FCM,检测Rb基因沉默后胆管癌细胞生长变化、细胞周期和细胞凋亡情况。RT-PCR和Western Blot检测Rb的mRNA和蛋白在胆管癌细胞系中的表达水平,同时检查与Rb相关的基因的表达情况。将QBC939细胞接种于裸鼠皮下,建立胆管癌裸鼠种植模型,观察体内种植瘤的发生率和生长情况。通过急性毒性试验找出TM5-FuNC对裸鼠的半数致死量(LD50)。根据急性毒性试验结果设定合适的药物剂量。MTT检测TM5-FuNC对转染psiRNARb-1前后QBC939细胞生长抑制的影响。将裸鼠模型再分成4组,分别为对照组,5-Fu组, TM5-FuNC组,psiRNARb-1和TM5-FuNC联合组。观察肿瘤体积变化,计算抑瘤率,绘制生长曲线。
实验结果
成功构建了针对Rb的真核表达载体,转染psiRNARb-1质粒48至72小时后QBC939细胞的生长相对于对照组细胞受到了一定程度的抑制,MTT实验结果显示当Rb基因在QBC939细胞中表达沉默时,对细胞生长有一定的抑制作用,在72小时内呈一定的时间依赖性。转染后QBC939细胞中Rb的mRNA表达相较于未转染细胞明显下调,Rb蛋白表达也明显受抑制。p16的mRNA和蛋白的表达变化情况与Rb相反。E2F1转录因子mRNA和蛋白的表达变化趋势与Rb相同。转染组中P27kip1蛋白的表达较未转染组有所增强。TM5-FuNC的急性动物实验中所有动物均未死亡,最大耐受剂量为1250 mg/kg。是临床给药剂量的5倍。TM5-FuNC 250和200mg/kg组的移植肿瘤体积在治疗后35天与对照组比较有显著差异(P<0.05)。电境结果显示,TM5-FuNC对荷瘤裸鼠的肿瘤细胞有诱导凋亡作用,且随着药物浓度的增大,细胞凋亡的形态改变越明显。转染psiRNARb-1的QBC939细胞可显著增强TM5-FuNC和5-Fu的生长抑制作用(P<0.05)。TM5-FuNC和TM5-FuNC+psiRNARb-1联合组与对照组和5-Fu组比较肿瘤体积的生长有明显差异(P<0.001)。TM5-FuNC和TM5-FuNC+psiRNARb-1联合组之间比较(P=0.44)有一定的差异显著性。
实验结论
1成功构建了4对针对Rb的siRNA真核表达载体,其中转染的psiRNARb-1质粒对胆管癌QBC939细胞有一定的抑制作用,并可诱导的胆管癌细胞G0/G1期阻滞和细胞凋亡。这种作用可能跟Rb与p16、E2F1、P27kip1之间的反馈调节机制相关。为我们将Rb作为基因治疗的靶目标提供了实验依据。
2通过急性毒性实验,我们获得了TM5-FuNC最大耐受剂量为后续实验提供了用药依据。
3 TM5-FuNC新剂型在内磁场的导向作用下能有效抑制人胆管癌裸鼠移植瘤的生长,使我们所研制的TM5-FuNC纳米药物制剂结合内磁场靶向治疗胆管癌的设计理念获得了实验的证实即可磁化的NiTi合金支架在保持胆道通畅引流的情况下,能高靶向的引导TM5-FuNC聚集在肿瘤部位实现对肿瘤高效地、持续地的治疗,并保证对机体的低毒性。
4 TM5-FuNC联合psiRNARb-1作用组相比单用TM5-FuNC组能更有效的抑制胆管癌肿瘤的生长,为TM5-FuNC联合基因治疗提供了理论依据和实验平台,为治疗中晚期胆管癌指明了一条新的途径。
Background & Objective: Cholangiocarcinoma is mainly named as the extrahepatic bile duct malignant tumor. The prognosis of it isn’t idealization. It’s scarce in epidemiology and is about 0.01%~0.5% in biopsy, about 3% in the total count of the malignant tumor. It’s about 0.07%~0.3% in domestic biopsy. The incidence rate of Cholangiocarcinoma is about 2.32% in all disease of biliary tract. The rate of female and male is 1.46:1. The incident rate is increasing progressively in recent years. The treatments of cholangiocarcinoma were given priority to operation or combine chemotherapy with radiation treatment. But the position of cholangiocarcinoma very deep and concealment in anatomy, it’s hard to detect cholangiocarcinoma in earlier period. As a result the effect of operation was discontentment, and the survival rate was very low. The chemiotherapy drug inhibit or kill tumor cells by act on different links of cell breeding, which can treat tumor. This is become a important method for clinical tumor treatment now. But the effect of chemiotherapy drug can not reach the aim of satisfactory. The report of the effect of 5-Fu is only reach 14%-18% regardless of simplex or combine medication.
Targeting treatment to malignant tumor is a new therapeutic method which as a result of conjunct nanotechology with chemotherapeutic drug rising in recent years. The principle is that the carrier including various kinds species group affinity carry particles and magnetic targeting particles take drug which could targeted reach to the locus of medication in order to treat disease. This will afford experctation for cure malignant tumor.
Our topic group lead by professor zoushengquan succeed get a great topic supported by the grant from The National Hi-Tech Research and Development Program of China (863 Program)(No. 2002AA214061). The core content of this project is that a magnetize cradle which be sacked by metal can reinforcement the TM5-FuNC targeting power while the cradle keeping bile duct drain smoothly. As a result that this will to achieve the real high selective and regional treatment. This will realizated break and new idea of Cholangiocarci- noma treatment, Furthermore, our group of the topic group has investigated deeply on the cholangiocarcinoma bionomics before. We concluded that there are complex network feed back mechanism between the oncogene and the anti-oncogene which is exist in the process of occurrence and develop of cholangiocarcinoma. pRb signal pathway were close importa- nt to the complex network feedback system of cholangiocarcinoma. It’s a great break in the research of the molecular cytogenetics in the cancer of extrahepatic biliary duct.
We based on the prophase research, put our emphasis on the pRb signal transduction pathway, and make some investigate on the apoptosis of the cholangiocarcinoma and the Fuction of the sensitization of the chemotherapy. We association the siRNA eukaryotic expression vector Rb with TM5-FuNC act on human cholangiocarcinoma cell in vivo and vitro, which will provid us a test base and realization pathway for the union opoerate of nanotechnology and gene therapy. Restricted to the time, we are limited and superficialis level, and we will make the Further step in the Future.
1. Methods and results: Eukaryotic expression vector of siRNA Rb were constructed, and used to transiently transfect to QBC939 mediated with LipofectamineTM 2000. The growth of QBC939 cell has been inhibited in some degree compare to the control group form 48h to 72h after transfection. Rb slience caused cell cycle arrest at G0/G1. Apoptotic cell index also increased in some degree. It show certain time dependency in 72h.Rb mRNA and protein under expression significantly compare to the control after transfection.p16 mRNA and protein expression inversed to Rb. E2F1 have identical change tendency with Rb. No animal dead in Acute toxicity test. The most tolerance dosage of TM5-FuNC is 1250mg/kg, which is 5 times dosage of clinical administration. High and middle dosage groups of TM5-FuNC have difference significantly compare to the control group (P <0.05) at 35 after treatment.The high and middle dosage groups of TM5-FuNC have discovered cells apoptosis under electron microscope. The ratio of growth inhibiting of TM5-FuNC and 5-Fu increased significantly after transfection. The tumor volume of TM5-FuNC group and psiRNARb-1+TM5-FuNC group have greatly difference compare with those of the control group and 5-Fu group (P<0.001). The tumor volume of TM5-FuNC group compare to tumor volume of TM5-FuNC group also have some difference (P=0.44).
Conclusions:
1 We are succeed constructed the eukaryotic expression vector of siRNA Rb which can specifically inhibit Rb expression There are certain inhibited growth effects of QBC939 cell line after transfecting psiRNARb-1 into cell. And Rb slience caused cell cycle arrest at G0/G1. Apoptotic cell index also increased in some degree, which may correlation with the Feedback adjust mechanism of Rb、p16、E2F1、P27kip1 protein . It can be used for later test.
2 We get TM5-FuNC the maximum tolerance dosage by acute toxicity experiment which is provided base for medication.
3 The TM5-FuNC can inhibit the tumor growth of human cholangiocarcinoma xeno- graft on nude mice. It show that our TM5-FuNC and design ideal of infra-magnetic field targeting get confirmation by test. The Ni-Ti alloy cradle who can be magnetized can high targeting led TM5-FuNC collecting in tumor tissue for treatment effectually and lastly when it keeping bile duct drain smoothly.
4 TM5-FuNC + psiRNARb-1 have better inhibit effect for cholangiocarcinoma compare to simplex TM5-FuNC. It would establish a test background for Further inosculatio TM5-FuNC with gene therapy.
引文
1. Vogelstein B,Kinzler KW.Cancer genes and the pathways they control.Nat Med, 2004 ,10(8): 789-799.
2. Albert B.Lowenfels,Patrick Maisonneuve.Epidemiology of biliary tract cancer:what is known, what is unknown?Biliary Tract Cancer International Workshop,2005,12(9):381-391
3. Zou SQ,Zhang L,Zen GZ.Clinical epidemiologic characteristics of 430 cases of gallbla-dder cancer.Chin Med J,1998,111(5):391-393.
4. Zou SQ,Zhang L.Relative risk factors analysis of 3922 cases of gallbladder cancer.China J Surg,2000,38(11):805-808.
5.黄志强.临床研究与新世纪胆道外科.第11届全国胆道外科学术会议论文编,2004,17-19.
6.邹声泉,龚建平主编.胆道肿瘤.外科学-前沿与争论,第一版.北京:人民卫生出版社,2003:600-607.
7. Zhao Y,Tan J,Zhuang L,et al.Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim.Proc Nat Acad Sci USA.2005 Nov 1;102(44):16090-16095.
8. Ohtani N,Yamakoshi K,Takahashi A,et al.The p16INK4a-RB pathway:molecular link between cellular senescence and tumor suppression.J Med Invest,2004 Aug;51(3-4):146-153.
9.秦军陈澍周施公胜等.p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
10.胡义利史光军李占武.p27KIP1和Rb蛋白在肝门部胆管癌组织中的表达及其意义.中华肝胆外科杂志.2002,8(8):507-508
11.Liu H,Dibling B,Spike B,et al.New roles for the RB tumor suppressor protein.Curr Opin Genet Dev. 2004 ,14(1):55-64.(26):15583-15586.
12.Dasgupta P,Betts V,Rastogi S,et al.Direct binding of apoptosis signal- regulating kinase-
1 to retinoblastoma protein:novel links between apoptotic signaling and cell cycle machinery .J Biol Chem.2004;279 (37): 8762-8769.
13.Mittnacht S. The retinoblastoma protein--from bench to bedside.Eur J Cell Biol.2005 Mar;84(2-3):97-107.
14.Larsen CJ. pRB, p53, p16INK4a, senescence and malignant transformation. Bull Cancer .2004 May;91(5):399-402.RJ,Garcia L,Baim DS,et al. Therapeutic Angiogenesis Using Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor Using Various Delivery Strategies. Curr Interv Cardiol Rep,1999,1(3):228-233.
15.Thongpraser S. The role of chemotherapy in cholangiocarcinoma. Annals of Oncology. 2005, 16 (l2): 93-96.
16.Sahoo SK,Labhasetwar V.Nanotech approaches to drug delivery and imaging. Drug Discov Today,2003,8(24):1112-1120.
1. Bernstein E,Caudy AA,Hammond SM,et al.Role for a bidentate ribonuclease in the initiation step of RNA inference. Nature ,2001,409:363-366
2. Hammond SM,Bernstein E,Beach D,et al. An RNA-directed nuclease mediates post- transcriptional gend silencing in Drosophila cells. Nature ,2000,404:293-296
3. Fire A,Xu S,Montgomery MK,et al. Potent and specific genetic interference by double stranded RNA in Caenorhabditis elegans. Nature,1998,391:806～811.
4. Martubez J, Patkaniowska A,Urlaub H,et al.Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell,2002,110(5):563-574
5. Elabshir J,Harborth SM ,Endeckel W,et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.Nature,2001,411:494-498
6. Schwarz D S, Hutvagner G, Haley B, Zamore PD. Evidence that siRNA Function as guides, not primers, in the Drosophila and human RNAi pathways. Mol Cell, 2002, 10(3): 537-548
7. Ramaswamy G, Slack FJ. siRNA: A guide for RNA silencing. Chem Biol. 2002 ;9(10): 1053-1055.
8. Lee RC, Ambros V. An extensive class of small RNAs in Caenorhabditis elegans.Science. 2001; 294 (5543): 862-864.
9. Khvorova A, Reynolds A, Jayasena SD. Functional siRNAs and miRNAs exhibit strand bias. Cell. 2003 Oct 17; 115(2): 209-216.
10. Ui-Tei K, Naito Y, Takahashi F, et al. Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res. 2004;32(3): 936-948.
11. Silva JM, Sachidanandam R, Hannon GJ. Free energy lights the path toward more effective RNAi. Nat Genet. 2003 Dec; 35(4): 303-305.
12. Reynolds A, Leake D, Boese Q, et al. Rational siRNA design for RNA interference. NatBiotechnol. 2004 Mar; 22(3): 326-330.
13. Miyagishi M,Taira K.U6 promoter driven siRNAs with four uridine 3′overhangs efficiently suppress targeted gene expression in mammalian cells.Nat Biotechnol.2002, 20 (5):497- 500.
14. Lee NS,Dohjima T,Bauer G,et al.Expression of small interfering RNAs targeted against HIV revtranscripts in human cells.Nat Biotechnol .2002,20(5):500 505.
15. Brummelkamp TR,Bernards R,Agami R.A system for stable expression of short interfering RNAs in mammalian cells.Science.2002,296(5567):550 553.
16. Yu JY,DeRuiter SL,Turner DL.RNA interference by expression of short interfering RNAs and hairpin RNAs in mammalian cells.Proc Natl Acad Sci USA.2002,99(9):6047 -6052.
17. Sui G,Soohoo C,Affarel B,Gay F,Shi Y,Forrester WC.A DNA vector-based RNAi technology to suppress gene express in mammalian cells.Proc Natl Acad Sci USA. 2002, 99:5515-5520
18. De Groen PC,Gores GJ,La Russo NF,et al. Biliary tract cancers .N Engl J Med,1999,341(18):1368-1378.
19. Fang X,Jin X,Xu HJ,et al. Expression of p16 induces transcriptionaldownregulation of the RB gene.Oncogene,1998,16(1):1-8.
20.秦军陈澍周施公胜等. p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
21.刘栋才,杨竹林,李永国.肝外胆管癌组织中细胞周期素D1、CDK4、Rb和p16的表达及其意义.中华实验外科杂志.2000,17(3):273
22. Li L,Yang T,Lian X.Effects of exogenous wild-type p16 gene transfection on the expression of cell cycle-related proteins in bladder cancer cell line.Cancer Invest,2005; 23(14):309-315
1.Vogelstein B,Kinzler KW.Cancer genes and the pathways they control.Nat Med, 2004 ,10 8): 789-799.
2.Albert B.Lowenfels,Patrick Maisonneuve.Epidemiology of biliary tract cancer:what is known,what is unknown?Biliary Tract Cancer International Workshop,2005,12(9):381-391
3.Zou SQ,Zhang L,Zen GZ.Clinical epidemiologic characteristics of 430 cases of gallbla- dder cancer.Chin Med J,1998,111(5):391-393.
4.Zou SQ,Zhang L.Relative risk factors analysis of 3922 cases of gallbladder cancer.China J Surg,2000,38(11):805-808.
5.黄志强.临床研究与新世纪胆道外科.第11届全国胆道外科学术会议论文编,2004,17-19.
6.邹声泉,龚建平主编.胆道肿瘤.外科学-前沿与争论,第一版.北京:人民卫生出版社,2003:600-607.
7.Zhao Y,Tan J,Zhuang L,et al.Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim.Proc Natl Acad Sci USA.2005 Nov 1;102(44):16090-16095.
8.Ohtani N,Yamakoshi K,Takahashi A,et al.The p16INK4a-RB pathway:molecular link between cellular senescence and tumor suppression.J Med Invest,2004 Aug;51(3-4):146-153.
9.秦军陈澍周施公胜等.p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
10.胡义利史光军李占武.p27KIP1和Rb蛋白在肝门部胆管癌组织中的表达及其意义.中华肝胆外科杂志.2002,8(8):507-508
11.Liu H,Dibling B,Spike B,et al.New roles for the RB tumor suppressor protein.Curr Opin Genet Dev. 2004 ,14(1):55-64.(26):15583-15586.
12.Dasgupta P,Betts V,Rastogi S,et al.Direct binding of apoptosis signal -regulating kinase 1 to retinoblastoma protein:novel links between apoptotic signaling and cell cycle machinery .J Biol Chem.2004;279 (37):8762-8769.
13.Mittnacht S. The retinoblastoma protein--from bench to bedside.Eur J Cell Biol.2005 Mar;84(2-3):97-107.
14.Larsen CJ.pRB,p53,p16INK4a,senescence and malignant transformation.Bull Cancer . 2004 May;91(5):399-402.
15.Simon B,Lubomierski N.Implication of the INK4a/ARF locus in gastroenteropancreatic neuroendocrine tumorigenesis.Ann N Y Acad Sci. 2004 Apr;1014:284-99.
16.Wang ZM,Yang H,Livingston DM.Endogenous E2F-1 promotes timely G0 exit of resting mouse embryo fibroblasts.Proc Natl Acad Sci USA,1998,95(26):15583- 15586.
17.Carlson CA,Ethier SP.Lack of RB potein correlates with increased sensitivity to UV radiation7 induced apoptosis in human breast cancer cells.Radiat Res,2000,154(5):590- 599.
18.Bramis J,Zacharatos P,Tsukamoto S,et al.E2F1 transcription factor immuno-expression is inversely associated with tumor growth in colon adenocarcinomas.Anticancer Res ,2004,24(5A):3041-3047
19.Polyak K, Kato JY, Solomon MJ, et al. P27Kip1, a cyclin-Cdk inhibitor,links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev 1994; 8:9-22
20.Toyoshima H, Hunter T. P27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21.Cell. 1994 Jul 15;78(1):67-74.
21.Karin Milde-Langosch, Maria Hagen, Ana-Maria.Expression and Prognostic Value of the Cell-cycle Regulatory Proteins, Rb, p16MTS1, p21WAF1, P27KIP1, Cyclin E, and Cyclin D2, in Ovarian Cancer. Int J Gynecol Pathol. 2003; 22(2): 168-174.
22.Masaharu Fukunaga.Immunohistochemical Characterization of Cyclin E and P27KIP1 Expression in Early Hydatidiform Moles. Int J Gynecol Pathol. 2004;23:259-264.
23.Runjan Chetty, MB ChB, FRCPath. P27 Protein and Cancers of the Gastrointestinal Tract and Liver. J Clin Gastroenterol 2003; 37(1):23-27.
24.Hayashi H, Ogawa N, Ishiwa N, et al. High cyclinE and low P27kip1 expressions are potentially poor prognostic factors in lung adenocarcinoma patients .Lung Cancer, 2001, 4(1):59-65
25.Edamoto Y, Hara A, Biernat W, etal. Alterations of RB1, p53 and Wnt pathways in hepatocellular carcinomasassociated with hepatitis C, hepatitis B and alcohol liver cirrhosis. Int Jcancer. 2003,106(3):334-341.
26.Sakatani T,Onyango P. Oncogenomics:prospects for the Future.Expert Rev Anticancer Ther. 2003;3(6):891-901.
27.Momparler RL. Cancer epigenetics.Oncogene, 2003,22(41):6479-6483.
28.Claudio PP,DeLuca A,Howard CM,et al.Function alanalysis of pRb/p130 interaction with cyclins.Cancer Res.1996,56(9):2003-2008.
29.He XS,Rong YH,Su Q,et al.Expression of p16 gene and Rb protein in gastric carcinoma and their clinicopathological significance.World J Gastroenterol.2005 Apr 21;11(15): 2218-2223.
30.Li L,Yang T. Effects of exogenous wild-type p16 gene transfection on the expression of cell cycle-related proteins in bladder cancer cell line.Cancer Invest,2005,23(14):309-315
31.杨贤东,肖晶,朱恩新.口腔鳞癌中Rb/cyclinD1/p16通路表达及病理意义.口腔医学研究.2005,21(3):295-287
32.Hui AM, Cui X. Makuuchi M, et al. Decreased p27(KipD ex-pr-ion and cyclin Dl over- expression.alone and in combination influence recurrence and survival of patients with resectable extra hepatic bile duct carcinoma. Hepatology,1999,30:1167-1173.
33.Lnda M,Cukor B,Tan SW, et al. Increased proteasome dependent degraduation of the cyclin dependent kinase inhibitor p27 in aggressive colorectal carcinomas. Nat Med, 1997,3(2): 231-234.
34.Derenzini M,Montanaro L,Vici1 M,et al.Relationship between the RB1 mRNA level and the expression of phosphorylated RB protein in human breast cancers: their rele- vance in cell proliferation activity and patient clinical outcome.Histol Histopathol. 2007,22(5):505 -513
1.石景生,王炳煌.胆道外科基础与临床(第一版).人民卫生出版社,2002年8月:674-691
2.Khan SA, Davidson RR, Goldin R, et al. Guidelines for the diagnosis and treatment of cholangiocarcinoma:censensus document . Gut.2002;51(supplVl):Vl1-Vl9
3.Rumalla,A, Peterson,B T.Diagnosis and therapy of biliary tract malignancy. Semin Gastrointest Drs .2000;11(3):168-173
4.徐萌主编.恶性肿瘤化疗及其对策(第一版)军事医学出版社,2002年9月:1013
5.Sanz-Altamira PM,Yermte K. Jekins RL. ET et al. A phase 11 o trial of 5-Fu, leurorin and Carboplattio in patients withunresectable biliary tree carcinoma. Cancer.1998;82: 2331-2335
6.Roco M.C.Nanotechnology:convergence with modern biology and medicine.Curr Opin Biotechnol.2003;14:337-346
7.周立国药物毒理学湖北科学技术出版社,2001年:25~26
8.化学药物急性毒性实验指导原则。国家食品药品监督管理局,2005.3
9.Thongpraser S. The role of chemotherapy in cholangiocarcinoma. Annals of Oncology. 2005, 16 (l2): 93-96.
10.Roco MC.Nanotechnology:convergence with modern biology and medicine. Curr Opin Biotechnol,2003,14(3):337-346.
11.Sahoo SK,Labhasetwar V.Nanotech approaches to drug delivery and imaging. Drug Discov Today,2003,8(24):1112-1120.
12.何跃明,吕新生,艾中立,等。纳米及微米磁微粒磁性顺铂微球对肝癌细胞的影响及机理.中国普通外科杂志.2004;13(3):179-184
1.Roco MC.Nanotechnology:convergence with modern biology and medicine. Curr Opin Biotechnol.2003;14(2):337-346
2.Pulfer SK,Ciccoto SL,Gallo JM.Distribution of small magnetic particles in brain tumorbearing rats.J Nurooncol.1999;4(2):99
3.Senyei AE, Reich SD,Goczy C,et al.In vivo kinetics of magnetically targeted low dose doxoruicin.J Pharm Sci,1981:70(4):389一391
4.吴远,王家陇,叶红军等.丝裂霉素C聚碳酸酯磁性微球的缓释功能及对移植性肝癌的靶向治疗作用.中华消化杂志.2001;21(11):691-693
5.Lind K, Kresse M,Debus NP,et al. A novel formulation for superparamagnetic iron oxide (SPIO) particles enhancing MR lymphography: comparison of physicochemical properties and the in vivo behaviour. J Drug Target 2002;10:221-230.
6.Jain S, Mishra V, Singh P, Dubey PK, Saraf DK, Vyas SP. RGD-anchored magnetic liposomes for monocytes/neutrophils-mediated brain targeting. Int J Pharm. 2003; 261:43-55.
7.Chen X, Hou Y, Tohme M, et al. Pegylated Arg-Gly-Asp peptide: 64Cu labeling and PET imaging of brain tumor alphavbeta3-integrin expression. J Nucl Med.2004; 45:1776- 1783.
8.任旭,王义录,李洪松等.逆行胰胆管造影对梗阻性黄疸的诊断价值.中华消化内镜杂志. 1996;13(1):17-19
9.张澍田,于中麟,于永征等.磁共振胰胆管造影术与直接造影术对照诊断胰胆系疾病.中华消化内镜杂志.1999,16(2):72-74
10.Wirleitner BB, Bitterl G, Bock G.7,8-Dihydroneopterin induced apoptosis in Jurkat T lymphocytes: a comparison with anti Fas-andhydrogen peroxide-mediated cell death. Biochempharm acol.1998;56(9):118
11.Muvaffak A, Gurhan I, Gunduz U, Hasirci N. Preparation and characterization of a biodegradable drug targeting system for anticancer drug delivery: microsphere- antibody conjugate. J Drug Target. 2005;13(1):151-159.
12.Liang LS, Wong W, Burt HM. Pharmacokinetic study of methotrexate following intra- articular injection of methotrexate loaded poly (L-lactic acid) microspheres in rabbits. J Pharm Sci.2005;94:1204-1215.
1.Thongpraser S. The role of chemotherapy in cholangiocarcinoma.Annals of Oncology. 2005; 16 (Suppl2): 93-96.
2.Harder J,Blum HE.Cholangiocarcinoma.Schweiz Rundsch Med Prax.2002,21;91(34):1352-1356
3.Coombes AG,Breeze V,Lin W,et al.Lacticacid stabilised albumin for microsphere for mulation and biomedical lcoatings.Biomaterials。2001,22(1):1-8
4.Dasgupta P,Betts V,Rastogi S,et al.Direct binding of apoptosis signal-regulating kinase 1 to retinoblastoma protein:novel links between apoptotic signaling and cell cycle machinery.J Biol Chem.2004 Sep 10;279(37):38762-8769.
5.Mittnacht S. The retinoblastoma protein--from bench to bedside.Eur J Cell Biol.2005 Mar;84(2-3):97-107.
6.Larsen CJ. pRB,p53,p16INK4a,senescence and malignant transformation.Bull Cancer. 2004 May;91(5):399-402.
7.Adams PD.Regulation of the retinoblastoma tumorsuppressor protein by cyclin/cdks. Biochim Biophys.2001;Acta 1471, M123-133.
8.Classon M.Harlow E.The retinoblastoma tumour suppressor in development and cancer. Nat Rev Cancer. 2002;2(1):910-917.
9.Derenzini M., Ceccarelli C., Santini D,et al.The prognostic value of the AgNOR parameter in human breast cancer depends on the ppRB and p53 status. J Clin Pathol. 2004;57:755-761.
10. Sherr CJ. McCormick F. . The RB and p53 pathways in cancer. Cancer Cell.(2002) 2, 103-112.
11.Semczuk A, Marzec B, Roessner A, et al. Loss of heterozygosity of the retinoblastoma gene is correlated with the altered ppRB expression in human endometrial cancer. Virchows Arch. 2002;441:577-583.
1.Brummelkamp TR, Bernards R, Agami R. Stable suppression of tumorigenicity by virus- mediated RNA interference. Cancer Cell. 2002 Sep; 2(3): 243-247.
2. Nagy P, Arndt-Jovin DJ, Jovin TM. Small interfering RNAs suppress the expression ofendogenous and GFP-fused epidermal growth factor receptor (erbB1) and induce apoptosis in erbB1-overexpressing cells. Exp Cell Res. 2003 Apr 15;285(1): 39-49.
3. Wilda M, Fuchs U, Wossmann W, et al. Killing of leukemic cells with BCR/ABL fusion gene by RNA interference (RNAi). Oncogene. 2002; 21(37):5716-5724.
4.Zhang L, Yang N, Mohamed-Hadley A, et al. Vector-based RNAi, a novel tool for isoform-specific knock-down of VEGF and anti-angiogenesis gene therapy of cancer. Biochem Biophys Res Commun. 2003 Apr 18; 303(4): 1169-1178.
5.Cioca DP, Aoki Y, Kiyosawa K. RNAi is a functional pathway with therapeutic potential in human myeloid leukemia cell lines. Cancer Gene Ther. 2003 Feb;10(2): 125-133.
6.Zhao Y,Tan J,Zhuang L,et al.Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim.Proc Natl Acad Sci USA.2005 Nov 1;102(44):16090-16095.
7.Ohtani N,Yamakoshi K,Takahashi A,et al.The p16INK4a-RB pathway:molecular link between cellular senescence and tumor suppression.J Med Invest,2004;51(3):146-153.
8.秦军陈澍周施公胜等.p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
9.胡义利史光军李占武.p27KIP1和Rb蛋白在肝门部胆管癌组织中的表达及其意义.中华肝胆外科杂志.2002,8(8):507-508
10.Pulfer S.K,Ciccoto S.L,Gallo J.M.Distribution of small magnetic particles in brain tumor-bearing rats.J Nurooncol,1999,4(2):99
11.吴远,王家陇,叶红军等.丝裂霉素C聚碳酸酯磁性微球的缓释功能及对移植性肝癌的靶向治疗作用.中华消化杂志,2001,21(11):691-693
12.Lind K, Kresse M,Debus NP,Muller RH.A novel formulation for superparamagnetic iron oxide (SPIO) particles enhancing MR lymphography: comparison of physicochemical properties and the in vivo behaviour. J Drug Target 2002;10:221-230.
13.Jain S, Mishra V, Singh P,et al. RGD-anchored magnetic liposomes for monocytes/ neutrophils-mediated brain targeting. Int J Pharm 2003;261:43-55.
14.Chen X, Hou Y, Tohme M, et al. Pegylated Arg-Gly-Asp peptide: 64Cu labeling and PET imaging of brain tumor alphavbeta3-integrin expression.Jnucl Med.2004;45: 1776-1783.
1.Bernstein E,Caudy AA,Hammond SM,et al.Role for a bidentate ribonuclease in the initiation step of RNA inference. Nature .2001,409:363-366
2.Hammond SM,Bernstein E,Beach D,et al. An RNA-directed nuclease mediates post- transcriptional gend silencing in Drosophila cells. Nature .2000;404:293-296
3.Colot V,Maloisel L,Rossignol JL.Interchromosomal transfer of epigenetic states inascobulus :Transfer of DNA methylation is mechanistically related to homologousrec ombination.Cell.1996;86:855-864.
4.Wassenegger M, Heimes S, Riedel L, et al. RNA-directed de novo methylation of genom- ic sequences in plants. Cell, 1994, 76: 567-576
5.Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993 Dec 3; 75(5): 843-854.
6.Reinhart BJ, Slack FJ, Basson M, et al. The 21-nucleotide let-7 RNA regulates develop- mental timing in Caenorhabditis elegans. Nature. 2000 Feb 24; 403(6772):901-906.
7. Grishok A, Pasquinelli AE, Conte D, et al. Genes and mechanisms related to RNA inter- ference regulate expression of the small temporal RNAs that control C. elegans develop- mental timing. Cell. 2001 Jul 13; 106(1): 23-34.
8.Hutvagner G, McLachlan J, Pasquinelli AE, A cellular function for the RNA-interference enzyme Dicer in the maturation of the let-7 small temporal RNA. Science. 2001 Aug 3; 293(5531): 834-838.
9. Doench JG, Petersen CP, Sharp PA. siRNAs can function as miRNAs. Genes Dev. 2003; Feb 15; 17(4): 438-442
10. Tabara H, Yigit E, Siomi H, et al. The dsRNA binding Protein RDE-4 interacts with RDE-l, DCR-I, and a DExH-box helicase to direct RNAi in C.elegans. Cell, 2002;28, 109(7): 861-871
11. Martubez J, Patkaniowska A,Urlaub H,et al.Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell,2002,110(5):563-574
12. Pairish S,Fleenor J,Xu S,et al.Functional amatpmy of a dsRNA trigger:Differental requirement for the to trigger strands in RNA interference.Mol Cell,6:1077-1087.
13. Martubez J, Patkaniowska A,Urlaub H,et al.Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell,2002,110(5):563-574
14. Elabshir J,Harborth SM ,Endeckel W,et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.Nature,2001,411:494-498
15. ]Schwarz D S, Hutvagner G, Haley B, Zamore PD. Evidence that siRNA function as guides, not primers, in the Drosophila and human RNAi pathways. Mol Cell, 2002, 10(3): 537-548
16. Ramaswamy G, Slack FJ. siRNA: A guide for RNA silencing. Chem Biol. 2002;9(10): 1053-1055.
17.Reynolds A, Leake D, Boese Q, et al. Rational siRNA design for RNA interference. Nat Biotechnol. 2004 Mar; 22(3): 326-330.
18. Brummelkamp,TR,Bemards R,Agami R. A system for stable expression of short inter- fering RNAs in mammalian cells.Science,2002,296:550-553.
19. Yu JY, DeRuiter SL. Turner DL. RNA interference by expression of shortinterfering RNAs and hairpin RNAs in mammalian cells .Proc Natl A cad Sci USA.2002;99(9): 6047-52.
20.Zamore PD,Tuschl T,Sharp PA,et al.RNAi:dsRNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals.Cell, 1999,101:25-33.
21.Song E, Lee SK, Wang J,et al. RNA interference targeting Fas protects mice from fulm- inant hepatitis. Nat Med, 2003,9(3):347-51
22.Caplen NJ,Fleenor J.Fire A,et al, dsRNA-mediated gene silencing in cultured Drosoph- ila cells :a tissue culture model for the analysis of RNA interference. Gene.2000;253:95
23.Elbashir SM, Harborth J, Weber K, Tuschl T. Analysis of gene function in somatic mam- malian cells using small interfering RNAs. Methods. 2002 Feb; 26(2): 199-213.
24.Paddison PJ,Caudy AA,Hannon GJ.Stable suppression of gene expression by RNAi in mammalian cells.Prpc Natl Acad Sci USA.2002,99(3):1443-1448.
25.Myers JW,Jones JT,Meyer T,et al.Recombinant Dicer efficiently converts large dsRNAs into siRNAs suitable for gene silencing.Nat Biotechnol.2003,21(3):324 328.
26.Miyagishi M.U6 promoter driven siRNAs with four uridine 3’overhangs efficiently supp ress targeted gene expression in mammalian cells.Nat Biotechnol.2002, 20 (5):497- 500.
27.Lee NS,Dohjima T,Bauer G,et al.Expression of small interfering RNAs targeted against HIV revtranscripts in human cells.Nat Biotechnol.2002,20(5):500 505.
28.Brummelkamp TR,Bernards R,Agami R.A system for stable expression of short interfer- ing RNAs in mammalian cells.Science.2002,296(5567):550-553.
29.Bettencourt R.Hemolin gene silencing by dsRNA injected into Cecropia pupae is to next generation embryos .Insect Molecular Biology ,2002,11(3):267-271.
30.Andrew G.Functional genomic analysis of C.elegans chromosome I by systematio RNA interference.Nature,2000,408:325-330.
31.Lee NS, Dohhjima T,Baner G,et al.Expression of small interference RNAs targeted agai- nst HIV-1 rev transcriptions in human cells. Nat Biotechnol,2002,20(19):500-505
32.Novina CD, Murray MF, Dykxhoom DM,et al. siRNA-directed inhibition of HIV-1 infe- ction. Nar Med.2002,8(15):681-686.
33. Kawasaki H, Taira K. Short hairpin type of dsRNAs that are controlled by tRNA(Val) promoter significantly induce RNAi-mediated gene silencing in the cytoplasm of human cells. Nucleic Acids Res.2003; 31(2):700-707.
34.Holle L, Hicks I。Song W, et al. Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro.Int J Oncol, 2004, 24(3):615-621..
35. Stege A, Priebsch A, Nieth C, et al. Stable and complete overcoming of MDRI/Pglycop rotein-mediated multidrug resistance in human gastric carcinoma cells by RNA interfere- nce. Cancer Gene Ther. 2004.11(11):699-706.
1. In Vivo Magnetic Resonance Detection of Cancer by Using Multifunctional Magnetic Nanocrystals.J Am Chem Soc. 2005 Sep 7;127(35):12387-91
2. Gangloff A,Hsueh WA,Kesner AL,et al. Estimation of Paclitaxel Biodistribution and Uptake in Human-Derived Xenografts In Vivo with 18F- Fluoropaclitaxel. J Nucl Med.2005 Nov;46(11):1866-71
3. Sun L,Fu BB,Liu DG. Systemic delivery of full-length C/EBP beta/liposome complex suppresses growth of human colon cancer in nude mice.Cell Res.2005;15(10):770-6.
4. Rong L,Wan YL,Liu YC,et al.Role of tissue factor in lung metastasis of colorectal carcinoma cells in vivo.Zhonghua Wei Chang Wai Ke Za Zhi.2005 Sep;8(5):447-50.
5. Shen J,Wang RF,Zhang CL,et al.Study on biodistribution and imaging of radioiodinated antisense oligonucleotides in nude mice bearing human lymphoma.Beijing Da Xue Xue Bao.2004 Dec;36(6):655-9.
6. Sagawa T,Takahashi M,Sato T,et al.Prolonged survival of mice with multiple liver metastases of human colon cancer by intravenous administration of replicable E1B- 55K-deleted adenovirus with E1A expressed by CEA promoter.Mol Ther.2004 Dec; 10(6):1043-50.
7. Chen X, Conti PS, Moats RA.In vivo near-infrared fluorescence imaging of integrin alphavbeta3 in brain tumor xenografts.Cancer Res.2004 Nov 1;64(21):8009-14.
8. van Furth WR,Laughlin S,Taylor MD.Imaging of murine brain tumors using a 1.5 Tesla clinical MRI system.Can J Neurol Sci.2003 Nov;30(4):326-32.
9. Bendel P,Koudinova N,Salomon Y.In vivo imaging of the neutron capture therapy agent BSH in mice using (10)B MRI.Magn Reson Med.2001 Jul;46(1):13-7.
2. Albert B.Lowenfels,Patrick Maisonneuve.Epidemiology of biliary tract cancer:what is known, what is unknown?Biliary Tract Cancer International Workshop,2005,12(9):381-391
3. Zou SQ,Zhang L,Zen GZ.Clinical epidemiologic characteristics of 430 cases of gallbla-dder cancer.Chin Med J,1998,111(5):391-393.
4. Zou SQ,Zhang L.Relative risk factors analysis of 3922 cases of gallbladder cancer.China J Surg,2000,38(11):805-808.
5.黄志强.临床研究与新世纪胆道外科.第11届全国胆道外科学术会议论文编,2004,17-19.
6.邹声泉,龚建平主编.胆道肿瘤.外科学-前沿与争论,第一版.北京:人民卫生出版社,2003:600-607.
7. Zhao Y,Tan J,Zhuang L,et al.Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim.Proc Nat Acad Sci USA.2005 Nov 1;102(44):16090-16095.
8. Ohtani N,Yamakoshi K,Takahashi A,et al.The p16INK4a-RB pathway:molecular link between cellular senescence and tumor suppression.J Med Invest,2004 Aug;51(3-4):146-153.
9.秦军陈澍周施公胜等.p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
10.胡义利史光军李占武.p27KIP1和Rb蛋白在肝门部胆管癌组织中的表达及其意义.中华肝胆外科杂志.2002,8(8):507-508
11.Liu H,Dibling B,Spike B,et al.New roles for the RB tumor suppressor protein.Curr Opin Genet Dev. 2004 ,14(1):55-64.(26):15583-15586.
12.Dasgupta P,Betts V,Rastogi S,et al.Direct binding of apoptosis signal- regulating kinase-
1 to retinoblastoma protein:novel links between apoptotic signaling and cell cycle machinery .J Biol Chem.2004;279 (37): 8762-8769.
13.Mittnacht S. The retinoblastoma protein--from bench to bedside.Eur J Cell Biol.2005 Mar;84(2-3):97-107.
14.Larsen CJ. pRB, p53, p16INK4a, senescence and malignant transformation. Bull Cancer .2004 May;91(5):399-402.RJ,Garcia L,Baim DS,et al. Therapeutic Angiogenesis Using Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor Using Various Delivery Strategies. Curr Interv Cardiol Rep,1999,1(3):228-233.
15.Thongpraser S. The role of chemotherapy in cholangiocarcinoma. Annals of Oncology. 2005, 16 (l2): 93-96.
16.Sahoo SK,Labhasetwar V.Nanotech approaches to drug delivery and imaging. Drug Discov Today,2003,8(24):1112-1120.
1. Bernstein E,Caudy AA,Hammond SM,et al.Role for a bidentate ribonuclease in the initiation step of RNA inference. Nature ,2001,409:363-366
2. Hammond SM,Bernstein E,Beach D,et al. An RNA-directed nuclease mediates post- transcriptional gend silencing in Drosophila cells. Nature ,2000,404:293-296
3. Fire A,Xu S,Montgomery MK,et al. Potent and specific genetic interference by double stranded RNA in Caenorhabditis elegans. Nature,1998,391:806～811.
4. Martubez J, Patkaniowska A,Urlaub H,et al.Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell,2002,110(5):563-574
5. Elabshir J,Harborth SM ,Endeckel W,et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.Nature,2001,411:494-498
6. Schwarz D S, Hutvagner G, Haley B, Zamore PD. Evidence that siRNA Function as guides, not primers, in the Drosophila and human RNAi pathways. Mol Cell, 2002, 10(3): 537-548
7. Ramaswamy G, Slack FJ. siRNA: A guide for RNA silencing. Chem Biol. 2002 ;9(10): 1053-1055.
8. Lee RC, Ambros V. An extensive class of small RNAs in Caenorhabditis elegans.Science. 2001; 294 (5543): 862-864.
9. Khvorova A, Reynolds A, Jayasena SD. Functional siRNAs and miRNAs exhibit strand bias. Cell. 2003 Oct 17; 115(2): 209-216.
10. Ui-Tei K, Naito Y, Takahashi F, et al. Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res. 2004;32(3): 936-948.
11. Silva JM, Sachidanandam R, Hannon GJ. Free energy lights the path toward more effective RNAi. Nat Genet. 2003 Dec; 35(4): 303-305.
12. Reynolds A, Leake D, Boese Q, et al. Rational siRNA design for RNA interference. NatBiotechnol. 2004 Mar; 22(3): 326-330.
13. Miyagishi M,Taira K.U6 promoter driven siRNAs with four uridine 3′overhangs efficiently suppress targeted gene expression in mammalian cells.Nat Biotechnol.2002, 20 (5):497- 500.
14. Lee NS,Dohjima T,Bauer G,et al.Expression of small interfering RNAs targeted against HIV revtranscripts in human cells.Nat Biotechnol .2002,20(5):500 505.
15. Brummelkamp TR,Bernards R,Agami R.A system for stable expression of short interfering RNAs in mammalian cells.Science.2002,296(5567):550 553.
16. Yu JY,DeRuiter SL,Turner DL.RNA interference by expression of short interfering RNAs and hairpin RNAs in mammalian cells.Proc Natl Acad Sci USA.2002,99(9):6047 -6052.
17. Sui G,Soohoo C,Affarel B,Gay F,Shi Y,Forrester WC.A DNA vector-based RNAi technology to suppress gene express in mammalian cells.Proc Natl Acad Sci USA. 2002, 99:5515-5520
18. De Groen PC,Gores GJ,La Russo NF,et al. Biliary tract cancers .N Engl J Med,1999,341(18):1368-1378.
19. Fang X,Jin X,Xu HJ,et al. Expression of p16 induces transcriptionaldownregulation of the RB gene.Oncogene,1998,16(1):1-8.
20.秦军陈澍周施公胜等. p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
21.刘栋才,杨竹林,李永国.肝外胆管癌组织中细胞周期素D1、CDK4、Rb和p16的表达及其意义.中华实验外科杂志.2000,17(3):273
22. Li L,Yang T,Lian X.Effects of exogenous wild-type p16 gene transfection on the expression of cell cycle-related proteins in bladder cancer cell line.Cancer Invest,2005; 23(14):309-315
1.Vogelstein B,Kinzler KW.Cancer genes and the pathways they control.Nat Med, 2004 ,10 8): 789-799.
2.Albert B.Lowenfels,Patrick Maisonneuve.Epidemiology of biliary tract cancer:what is known,what is unknown?Biliary Tract Cancer International Workshop,2005,12(9):381-391
3.Zou SQ,Zhang L,Zen GZ.Clinical epidemiologic characteristics of 430 cases of gallbla- dder cancer.Chin Med J,1998,111(5):391-393.
4.Zou SQ,Zhang L.Relative risk factors analysis of 3922 cases of gallbladder cancer.China J Surg,2000,38(11):805-808.
5.黄志强.临床研究与新世纪胆道外科.第11届全国胆道外科学术会议论文编,2004,17-19.
6.邹声泉,龚建平主编.胆道肿瘤.外科学-前沿与争论,第一版.北京:人民卫生出版社,2003:600-607.
7.Zhao Y,Tan J,Zhuang L,et al.Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim.Proc Natl Acad Sci USA.2005 Nov 1;102(44):16090-16095.
8.Ohtani N,Yamakoshi K,Takahashi A,et al.The p16INK4a-RB pathway:molecular link between cellular senescence and tumor suppression.J Med Invest,2004 Aug;51(3-4):146-153.
9.秦军陈澍周施公胜等.p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
10.胡义利史光军李占武.p27KIP1和Rb蛋白在肝门部胆管癌组织中的表达及其意义.中华肝胆外科杂志.2002,8(8):507-508
11.Liu H,Dibling B,Spike B,et al.New roles for the RB tumor suppressor protein.Curr Opin Genet Dev. 2004 ,14(1):55-64.(26):15583-15586.
12.Dasgupta P,Betts V,Rastogi S,et al.Direct binding of apoptosis signal -regulating kinase 1 to retinoblastoma protein:novel links between apoptotic signaling and cell cycle machinery .J Biol Chem.2004;279 (37):8762-8769.
13.Mittnacht S. The retinoblastoma protein--from bench to bedside.Eur J Cell Biol.2005 Mar;84(2-3):97-107.
14.Larsen CJ.pRB,p53,p16INK4a,senescence and malignant transformation.Bull Cancer . 2004 May;91(5):399-402.
15.Simon B,Lubomierski N.Implication of the INK4a/ARF locus in gastroenteropancreatic neuroendocrine tumorigenesis.Ann N Y Acad Sci. 2004 Apr;1014:284-99.
16.Wang ZM,Yang H,Livingston DM.Endogenous E2F-1 promotes timely G0 exit of resting mouse embryo fibroblasts.Proc Natl Acad Sci USA,1998,95(26):15583- 15586.
17.Carlson CA,Ethier SP.Lack of RB potein correlates with increased sensitivity to UV radiation7 induced apoptosis in human breast cancer cells.Radiat Res,2000,154(5):590- 599.
18.Bramis J,Zacharatos P,Tsukamoto S,et al.E2F1 transcription factor immuno-expression is inversely associated with tumor growth in colon adenocarcinomas.Anticancer Res ,2004,24(5A):3041-3047
19.Polyak K, Kato JY, Solomon MJ, et al. P27Kip1, a cyclin-Cdk inhibitor,links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev 1994; 8:9-22
20.Toyoshima H, Hunter T. P27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21.Cell. 1994 Jul 15;78(1):67-74.
21.Karin Milde-Langosch, Maria Hagen, Ana-Maria.Expression and Prognostic Value of the Cell-cycle Regulatory Proteins, Rb, p16MTS1, p21WAF1, P27KIP1, Cyclin E, and Cyclin D2, in Ovarian Cancer. Int J Gynecol Pathol. 2003; 22(2): 168-174.
22.Masaharu Fukunaga.Immunohistochemical Characterization of Cyclin E and P27KIP1 Expression in Early Hydatidiform Moles. Int J Gynecol Pathol. 2004;23:259-264.
23.Runjan Chetty, MB ChB, FRCPath. P27 Protein and Cancers of the Gastrointestinal Tract and Liver. J Clin Gastroenterol 2003; 37(1):23-27.
24.Hayashi H, Ogawa N, Ishiwa N, et al. High cyclinE and low P27kip1 expressions are potentially poor prognostic factors in lung adenocarcinoma patients .Lung Cancer, 2001, 4(1):59-65
25.Edamoto Y, Hara A, Biernat W, etal. Alterations of RB1, p53 and Wnt pathways in hepatocellular carcinomasassociated with hepatitis C, hepatitis B and alcohol liver cirrhosis. Int Jcancer. 2003,106(3):334-341.
26.Sakatani T,Onyango P. Oncogenomics:prospects for the Future.Expert Rev Anticancer Ther. 2003;3(6):891-901.
27.Momparler RL. Cancer epigenetics.Oncogene, 2003,22(41):6479-6483.
28.Claudio PP,DeLuca A,Howard CM,et al.Function alanalysis of pRb/p130 interaction with cyclins.Cancer Res.1996,56(9):2003-2008.
29.He XS,Rong YH,Su Q,et al.Expression of p16 gene and Rb protein in gastric carcinoma and their clinicopathological significance.World J Gastroenterol.2005 Apr 21;11(15): 2218-2223.
30.Li L,Yang T. Effects of exogenous wild-type p16 gene transfection on the expression of cell cycle-related proteins in bladder cancer cell line.Cancer Invest,2005,23(14):309-315
31.杨贤东,肖晶,朱恩新.口腔鳞癌中Rb/cyclinD1/p16通路表达及病理意义.口腔医学研究.2005,21(3):295-287
32.Hui AM, Cui X. Makuuchi M, et al. Decreased p27(KipD ex-pr-ion and cyclin Dl over- expression.alone and in combination influence recurrence and survival of patients with resectable extra hepatic bile duct carcinoma. Hepatology,1999,30:1167-1173.
33.Lnda M,Cukor B,Tan SW, et al. Increased proteasome dependent degraduation of the cyclin dependent kinase inhibitor p27 in aggressive colorectal carcinomas. Nat Med, 1997,3(2): 231-234.
34.Derenzini M,Montanaro L,Vici1 M,et al.Relationship between the RB1 mRNA level and the expression of phosphorylated RB protein in human breast cancers: their rele- vance in cell proliferation activity and patient clinical outcome.Histol Histopathol. 2007,22(5):505 -513
1.石景生,王炳煌.胆道外科基础与临床(第一版).人民卫生出版社,2002年8月:674-691
2.Khan SA, Davidson RR, Goldin R, et al. Guidelines for the diagnosis and treatment of cholangiocarcinoma:censensus document . Gut.2002;51(supplVl):Vl1-Vl9
3.Rumalla,A, Peterson,B T.Diagnosis and therapy of biliary tract malignancy. Semin Gastrointest Drs .2000;11(3):168-173
4.徐萌主编.恶性肿瘤化疗及其对策(第一版)军事医学出版社,2002年9月:1013
5.Sanz-Altamira PM,Yermte K. Jekins RL. ET et al. A phase 11 o trial of 5-Fu, leurorin and Carboplattio in patients withunresectable biliary tree carcinoma. Cancer.1998;82: 2331-2335
6.Roco M.C.Nanotechnology:convergence with modern biology and medicine.Curr Opin Biotechnol.2003;14:337-346
7.周立国药物毒理学湖北科学技术出版社,2001年:25~26
8.化学药物急性毒性实验指导原则。国家食品药品监督管理局,2005.3
9.Thongpraser S. The role of chemotherapy in cholangiocarcinoma. Annals of Oncology. 2005, 16 (l2): 93-96.
10.Roco MC.Nanotechnology:convergence with modern biology and medicine. Curr Opin Biotechnol,2003,14(3):337-346.
11.Sahoo SK,Labhasetwar V.Nanotech approaches to drug delivery and imaging. Drug Discov Today,2003,8(24):1112-1120.
12.何跃明,吕新生,艾中立,等。纳米及微米磁微粒磁性顺铂微球对肝癌细胞的影响及机理.中国普通外科杂志.2004;13(3):179-184
1.Roco MC.Nanotechnology:convergence with modern biology and medicine. Curr Opin Biotechnol.2003;14(2):337-346
2.Pulfer SK,Ciccoto SL,Gallo JM.Distribution of small magnetic particles in brain tumorbearing rats.J Nurooncol.1999;4(2):99
3.Senyei AE, Reich SD,Goczy C,et al.In vivo kinetics of magnetically targeted low dose doxoruicin.J Pharm Sci,1981:70(4):389一391
4.吴远,王家陇,叶红军等.丝裂霉素C聚碳酸酯磁性微球的缓释功能及对移植性肝癌的靶向治疗作用.中华消化杂志.2001;21(11):691-693
5.Lind K, Kresse M,Debus NP,et al. A novel formulation for superparamagnetic iron oxide (SPIO) particles enhancing MR lymphography: comparison of physicochemical properties and the in vivo behaviour. J Drug Target 2002;10:221-230.
6.Jain S, Mishra V, Singh P, Dubey PK, Saraf DK, Vyas SP. RGD-anchored magnetic liposomes for monocytes/neutrophils-mediated brain targeting. Int J Pharm. 2003; 261:43-55.
7.Chen X, Hou Y, Tohme M, et al. Pegylated Arg-Gly-Asp peptide: 64Cu labeling and PET imaging of brain tumor alphavbeta3-integrin expression. J Nucl Med.2004; 45:1776- 1783.
8.任旭,王义录,李洪松等.逆行胰胆管造影对梗阻性黄疸的诊断价值.中华消化内镜杂志. 1996;13(1):17-19
9.张澍田,于中麟,于永征等.磁共振胰胆管造影术与直接造影术对照诊断胰胆系疾病.中华消化内镜杂志.1999,16(2):72-74
10.Wirleitner BB, Bitterl G, Bock G.7,8-Dihydroneopterin induced apoptosis in Jurkat T lymphocytes: a comparison with anti Fas-andhydrogen peroxide-mediated cell death. Biochempharm acol.1998;56(9):118
11.Muvaffak A, Gurhan I, Gunduz U, Hasirci N. Preparation and characterization of a biodegradable drug targeting system for anticancer drug delivery: microsphere- antibody conjugate. J Drug Target. 2005;13(1):151-159.
12.Liang LS, Wong W, Burt HM. Pharmacokinetic study of methotrexate following intra- articular injection of methotrexate loaded poly (L-lactic acid) microspheres in rabbits. J Pharm Sci.2005;94:1204-1215.
1.Thongpraser S. The role of chemotherapy in cholangiocarcinoma.Annals of Oncology. 2005; 16 (Suppl2): 93-96.
2.Harder J,Blum HE.Cholangiocarcinoma.Schweiz Rundsch Med Prax.2002,21;91(34):1352-1356
3.Coombes AG,Breeze V,Lin W,et al.Lacticacid stabilised albumin for microsphere for mulation and biomedical lcoatings.Biomaterials。2001,22(1):1-8
4.Dasgupta P,Betts V,Rastogi S,et al.Direct binding of apoptosis signal-regulating kinase 1 to retinoblastoma protein:novel links between apoptotic signaling and cell cycle machinery.J Biol Chem.2004 Sep 10;279(37):38762-8769.
5.Mittnacht S. The retinoblastoma protein--from bench to bedside.Eur J Cell Biol.2005 Mar;84(2-3):97-107.
6.Larsen CJ. pRB,p53,p16INK4a,senescence and malignant transformation.Bull Cancer. 2004 May;91(5):399-402.
7.Adams PD.Regulation of the retinoblastoma tumorsuppressor protein by cyclin/cdks. Biochim Biophys.2001;Acta 1471, M123-133.
8.Classon M.Harlow E.The retinoblastoma tumour suppressor in development and cancer. Nat Rev Cancer. 2002;2(1):910-917.
9.Derenzini M., Ceccarelli C., Santini D,et al.The prognostic value of the AgNOR parameter in human breast cancer depends on the ppRB and p53 status. J Clin Pathol. 2004;57:755-761.
10. Sherr CJ. McCormick F. . The RB and p53 pathways in cancer. Cancer Cell.(2002) 2, 103-112.
11.Semczuk A, Marzec B, Roessner A, et al. Loss of heterozygosity of the retinoblastoma gene is correlated with the altered ppRB expression in human endometrial cancer. Virchows Arch. 2002;441:577-583.
1.Brummelkamp TR, Bernards R, Agami R. Stable suppression of tumorigenicity by virus- mediated RNA interference. Cancer Cell. 2002 Sep; 2(3): 243-247.
2. Nagy P, Arndt-Jovin DJ, Jovin TM. Small interfering RNAs suppress the expression ofendogenous and GFP-fused epidermal growth factor receptor (erbB1) and induce apoptosis in erbB1-overexpressing cells. Exp Cell Res. 2003 Apr 15;285(1): 39-49.
3. Wilda M, Fuchs U, Wossmann W, et al. Killing of leukemic cells with BCR/ABL fusion gene by RNA interference (RNAi). Oncogene. 2002; 21(37):5716-5724.
4.Zhang L, Yang N, Mohamed-Hadley A, et al. Vector-based RNAi, a novel tool for isoform-specific knock-down of VEGF and anti-angiogenesis gene therapy of cancer. Biochem Biophys Res Commun. 2003 Apr 18; 303(4): 1169-1178.
5.Cioca DP, Aoki Y, Kiyosawa K. RNAi is a functional pathway with therapeutic potential in human myeloid leukemia cell lines. Cancer Gene Ther. 2003 Feb;10(2): 125-133.
6.Zhao Y,Tan J,Zhuang L,et al.Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim.Proc Natl Acad Sci USA.2005 Nov 1;102(44):16090-16095.
7.Ohtani N,Yamakoshi K,Takahashi A,et al.The p16INK4a-RB pathway:molecular link between cellular senescence and tumor suppression.J Med Invest,2004;51(3):146-153.
8.秦军陈澍周施公胜等.p16和Rb在胆道良恶性病变中的表达及其意义.中国肿瘤临床与康复.2004,11(5):392-394
9.胡义利史光军李占武.p27KIP1和Rb蛋白在肝门部胆管癌组织中的表达及其意义.中华肝胆外科杂志.2002,8(8):507-508
10.Pulfer S.K,Ciccoto S.L,Gallo J.M.Distribution of small magnetic particles in brain tumor-bearing rats.J Nurooncol,1999,4(2):99
11.吴远,王家陇,叶红军等.丝裂霉素C聚碳酸酯磁性微球的缓释功能及对移植性肝癌的靶向治疗作用.中华消化杂志,2001,21(11):691-693
12.Lind K, Kresse M,Debus NP,Muller RH.A novel formulation for superparamagnetic iron oxide (SPIO) particles enhancing MR lymphography: comparison of physicochemical properties and the in vivo behaviour. J Drug Target 2002;10:221-230.
13.Jain S, Mishra V, Singh P,et al. RGD-anchored magnetic liposomes for monocytes/ neutrophils-mediated brain targeting. Int J Pharm 2003;261:43-55.
14.Chen X, Hou Y, Tohme M, et al. Pegylated Arg-Gly-Asp peptide: 64Cu labeling and PET imaging of brain tumor alphavbeta3-integrin expression.Jnucl Med.2004;45: 1776-1783.
1.Bernstein E,Caudy AA,Hammond SM,et al.Role for a bidentate ribonuclease in the initiation step of RNA inference. Nature .2001,409:363-366
2.Hammond SM,Bernstein E,Beach D,et al. An RNA-directed nuclease mediates post- transcriptional gend silencing in Drosophila cells. Nature .2000;404:293-296
3.Colot V,Maloisel L,Rossignol JL.Interchromosomal transfer of epigenetic states inascobulus :Transfer of DNA methylation is mechanistically related to homologousrec ombination.Cell.1996;86:855-864.
4.Wassenegger M, Heimes S, Riedel L, et al. RNA-directed de novo methylation of genom- ic sequences in plants. Cell, 1994, 76: 567-576
5.Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993 Dec 3; 75(5): 843-854.
6.Reinhart BJ, Slack FJ, Basson M, et al. The 21-nucleotide let-7 RNA regulates develop- mental timing in Caenorhabditis elegans. Nature. 2000 Feb 24; 403(6772):901-906.
7. Grishok A, Pasquinelli AE, Conte D, et al. Genes and mechanisms related to RNA inter- ference regulate expression of the small temporal RNAs that control C. elegans develop- mental timing. Cell. 2001 Jul 13; 106(1): 23-34.
8.Hutvagner G, McLachlan J, Pasquinelli AE, A cellular function for the RNA-interference enzyme Dicer in the maturation of the let-7 small temporal RNA. Science. 2001 Aug 3; 293(5531): 834-838.
9. Doench JG, Petersen CP, Sharp PA. siRNAs can function as miRNAs. Genes Dev. 2003; Feb 15; 17(4): 438-442
10. Tabara H, Yigit E, Siomi H, et al. The dsRNA binding Protein RDE-4 interacts with RDE-l, DCR-I, and a DExH-box helicase to direct RNAi in C.elegans. Cell, 2002;28, 109(7): 861-871
11. Martubez J, Patkaniowska A,Urlaub H,et al.Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell,2002,110(5):563-574
12. Pairish S,Fleenor J,Xu S,et al.Functional amatpmy of a dsRNA trigger:Differental requirement for the to trigger strands in RNA interference.Mol Cell,6:1077-1087.
13. Martubez J, Patkaniowska A,Urlaub H,et al.Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell,2002,110(5):563-574
14. Elabshir J,Harborth SM ,Endeckel W,et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.Nature,2001,411:494-498
15. ]Schwarz D S, Hutvagner G, Haley B, Zamore PD. Evidence that siRNA function as guides, not primers, in the Drosophila and human RNAi pathways. Mol Cell, 2002, 10(3): 537-548
16. Ramaswamy G, Slack FJ. siRNA: A guide for RNA silencing. Chem Biol. 2002;9(10): 1053-1055.
17.Reynolds A, Leake D, Boese Q, et al. Rational siRNA design for RNA interference. Nat Biotechnol. 2004 Mar; 22(3): 326-330.
18. Brummelkamp,TR,Bemards R,Agami R. A system for stable expression of short inter- fering RNAs in mammalian cells.Science,2002,296:550-553.
19. Yu JY, DeRuiter SL. Turner DL. RNA interference by expression of shortinterfering RNAs and hairpin RNAs in mammalian cells .Proc Natl A cad Sci USA.2002;99(9): 6047-52.
20.Zamore PD,Tuschl T,Sharp PA,et al.RNAi:dsRNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals.Cell, 1999,101:25-33.
21.Song E, Lee SK, Wang J,et al. RNA interference targeting Fas protects mice from fulm- inant hepatitis. Nat Med, 2003,9(3):347-51
22.Caplen NJ,Fleenor J.Fire A,et al, dsRNA-mediated gene silencing in cultured Drosoph- ila cells :a tissue culture model for the analysis of RNA interference. Gene.2000;253:95
23.Elbashir SM, Harborth J, Weber K, Tuschl T. Analysis of gene function in somatic mam- malian cells using small interfering RNAs. Methods. 2002 Feb; 26(2): 199-213.
24.Paddison PJ,Caudy AA,Hannon GJ.Stable suppression of gene expression by RNAi in mammalian cells.Prpc Natl Acad Sci USA.2002,99(3):1443-1448.
25.Myers JW,Jones JT,Meyer T,et al.Recombinant Dicer efficiently converts large dsRNAs into siRNAs suitable for gene silencing.Nat Biotechnol.2003,21(3):324 328.
26.Miyagishi M.U6 promoter driven siRNAs with four uridine 3’overhangs efficiently supp ress targeted gene expression in mammalian cells.Nat Biotechnol.2002, 20 (5):497- 500.
27.Lee NS,Dohjima T,Bauer G,et al.Expression of small interfering RNAs targeted against HIV revtranscripts in human cells.Nat Biotechnol.2002,20(5):500 505.
28.Brummelkamp TR,Bernards R,Agami R.A system for stable expression of short interfer- ing RNAs in mammalian cells.Science.2002,296(5567):550-553.
29.Bettencourt R.Hemolin gene silencing by dsRNA injected into Cecropia pupae is to next generation embryos .Insect Molecular Biology ,2002,11(3):267-271.
30.Andrew G.Functional genomic analysis of C.elegans chromosome I by systematio RNA interference.Nature,2000,408:325-330.
31.Lee NS, Dohhjima T,Baner G,et al.Expression of small interference RNAs targeted agai- nst HIV-1 rev transcriptions in human cells. Nat Biotechnol,2002,20(19):500-505
32.Novina CD, Murray MF, Dykxhoom DM,et al. siRNA-directed inhibition of HIV-1 infe- ction. Nar Med.2002,8(15):681-686.
33. Kawasaki H, Taira K. Short hairpin type of dsRNAs that are controlled by tRNA(Val) promoter significantly induce RNAi-mediated gene silencing in the cytoplasm of human cells. Nucleic Acids Res.2003; 31(2):700-707.
34.Holle L, Hicks I。Song W, et al. Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro.Int J Oncol, 2004, 24(3):615-621..
35. Stege A, Priebsch A, Nieth C, et al. Stable and complete overcoming of MDRI/Pglycop rotein-mediated multidrug resistance in human gastric carcinoma cells by RNA interfere- nce. Cancer Gene Ther. 2004.11(11):699-706.
1. In Vivo Magnetic Resonance Detection of Cancer by Using Multifunctional Magnetic Nanocrystals.J Am Chem Soc. 2005 Sep 7;127(35):12387-91
2. Gangloff A,Hsueh WA,Kesner AL,et al. Estimation of Paclitaxel Biodistribution and Uptake in Human-Derived Xenografts In Vivo with 18F- Fluoropaclitaxel. J Nucl Med.2005 Nov;46(11):1866-71
3. Sun L,Fu BB,Liu DG. Systemic delivery of full-length C/EBP beta/liposome complex suppresses growth of human colon cancer in nude mice.Cell Res.2005;15(10):770-6.
4. Rong L,Wan YL,Liu YC,et al.Role of tissue factor in lung metastasis of colorectal carcinoma cells in vivo.Zhonghua Wei Chang Wai Ke Za Zhi.2005 Sep;8(5):447-50.
5. Shen J,Wang RF,Zhang CL,et al.Study on biodistribution and imaging of radioiodinated antisense oligonucleotides in nude mice bearing human lymphoma.Beijing Da Xue Xue Bao.2004 Dec;36(6):655-9.
6. Sagawa T,Takahashi M,Sato T,et al.Prolonged survival of mice with multiple liver metastases of human colon cancer by intravenous administration of replicable E1B- 55K-deleted adenovirus with E1A expressed by CEA promoter.Mol Ther.2004 Dec; 10(6):1043-50.
7. Chen X, Conti PS, Moats RA.In vivo near-infrared fluorescence imaging of integrin alphavbeta3 in brain tumor xenografts.Cancer Res.2004 Nov 1;64(21):8009-14.
8. van Furth WR,Laughlin S,Taylor MD.Imaging of murine brain tumors using a 1.5 Tesla clinical MRI system.Can J Neurol Sci.2003 Nov;30(4):326-32.
9. Bendel P,Koudinova N,Salomon Y.In vivo imaging of the neutron capture therapy agent BSH in mice using (10)B MRI.Magn Reson Med.2001 Jul;46(1):13-7.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |