RNAi抑制人前列腺癌PC3细胞EphB4基因表达的实验研究  

Inhibitory effect of RNAi on the expression of EphB4 gene in human prostate cancer PC3 cells

作　　者：方晓亮[1] 潘骏[2] 顾正勤[1] 康健[1] 齐隽[1] 

机构地区：[1]上海交通大学医学院附属新华医院泌尿外科,上海200092 [2]上海市儿科医学研究所

出　　处：《中国男科学杂志》2009年第6期6-10,共5页Chinese Journal of Andrology

基　　金：上海市科委重点项目基金(06JC14086);上海交通大学医学院附属新华医院院级科技基金(07YJ02)

摘　　要：目的观察RNAi对人前列腺癌PC3细胞EphB4基因表达的抑制效应,寻求高效率的干扰片段。方法设计并化学合成特异性针对EphB4基因的siRNA(siRNA-1、siRNA-2、siRNA-3)为实验组,另设一无意义siRNA为对照组,脂质体转染PC3细胞。荧光显微镜观察siRNA的转染效率;分别应用RT-PCR以及Western Blot、免疫荧光组织化学染色方法检测EphB4 mRNA和蛋白的表达,以及应用MTT法检测siRNA对PC3细胞生长的影响,比较实验组与对照组的差异及评估siRNA阻抑效应。结果实验组siRNA对PC3细胞EphB4 mRNA和蛋白的表达较对照组均有不同程度的下降,统计学分析其差异具有统计学意义(P<0.05)。siRNA对PC3细胞的生长具有抑制作用。结论RNAi可以有效地抑制人前列腺癌PC3细胞EphB4的表达,找到了具有较高抑制效率的siRNA,为进一步探究EphB4的生物学功能、体内实验和临床抗肿瘤药物研究打下基础。Objective To assess the expression state of EphB4 gene in human prostate cancer PC3 cells transfected by EphB4-specific siRNAs, and screen the optimal siRNA（s） sequence segment for the greatest suppressing state. Methods EphB4-specific siRNA was chemically synthesized and transfected into the PC3 ceils by liposome. The transfection efficiency of siRNA was detected with fluorescence microscopy; RT-PCR and western blot, immtmohistochemical methods were respectively applied respectively to detect the EphB4 mRNA and protein expression, MTT was used to detect the growth of PC3 cells under pre and post transfection with EphB4-specific siRNA,A nonsense siRNA was set as control. Results Compared with that in the control group, the level of mRNA or protein expression of EphB4 decreased significantly in transfected with EphB4-specific siRNA, which was statistically significant （P〈0.05）. The proliferation of PC3 cells was found to be inhibited after transfection with EphB4-specific siRNA. Conclusion The EphB4- specific siRNA could significantly inhibited the expression of EphB4 gene in prostate cancer PC3cells, and the optimal siRNA（s） sequence segment was identified. It will be used for function explore of EphB4 gene and discovery new antitumor drug in the future researches.

关 键 词：RNA干扰 前列肿瘤 EphB4基因 

分 类 号：R737.25[医药卫生—肿瘤]