RNAi抑制膀胱癌T24细胞中VEGF.docVIP

RNA干扰对膀胱癌T24细胞血管内皮生长因子C表达的影响 祖雄兵 叶章群 周四维 齐琳 张向阳 [摘要] 目的 观察RNA干扰（RNAi）抑制膀胱癌T24细胞中血管内皮生长因子C（VEGF-C）表达和提高化疗敏感性。方法 根据转染效率最高时pEGFPN1质粒与脂质体的比例，转染T24细胞，实验分为3组，RT-PCR检测VEGF-CmRNA的表达，流式细胞仪分析细胞凋亡率。结果　转染后24h，即有mRNA表达水平的下降，Quantity one半定量：24小时0.57±0.17，48小时0.42±0.11，72小时0.33±0.09。VEGF-C抑制后，吉西他滨诱导的凋亡率明显升高，转染组为(41.38±1.54)%，明显高于未转染组(22.87±1.40)%与脂质体组(23.47±1.58)% (P0.05)。结论 RNAi可高效稳定的抑制膀胱癌T24细胞中VEGF-C的表达，明显提高了吉西他滨诱导膀胱癌T24细胞凋亡的药物敏感性。 [关键词] 膀胱癌；VEGF-C；RNA干扰；化学治疗；细胞凋亡 RNA interference on Vascular endothelial growth factor C mRNA expression in bladder cancer T24 cells 　ZU Xiong-bing﹡, YE Zhang-qun, ZHOU Si-wei, QI Lin,ZHANG Xiangyang. *Department of Urology,Xiangya Hospital,Central South Universiy,Changsha 410008, China [Abstract] Objective To evaluate the improvement of sensitivity to chemotherapeutical drugs in T24 cell line of bladder cancer when the expression of VEGF-C has been inhibited by RNA interference (RNAi). Method We constructed RNA interference vectors with H1 promoter against VEGF-C and then transferred them into the T24 cell line. the expression of VEGF-C was determined by RT-PCR, and FCM was employed to examine the apoptosis index of T24 cell line induced by Gemcitabine. Results The expression level of mRNA declined after transfection. Analyzed with Quantity One, the data of semi-quantitative determination shows 0.57?0.17 after 24hs, 0.42?0.11 after 48hs, 0.33?0.09 after 72hs. The apoptosis index was (41.38?1.54)% in the transferred group, (22.87?1.40)%in the control group and (23.47?1.58)% in the liposome group. The sensitivity of T24 cells to gemcitabine increased after VEGF-C had been inhibited and the apoptosis index examined by FCM was significantly increased (P0.01). The expression level of VEGF-C has negative correlation with the apoptosis index (P0.05). Conclusion The small Hairpin siRNA produced by RNAi vectors with H1 promoter can effectively suppress the 基金项目: 国家自然科学基金资助项目；中国博士后科学基金资助项目（2005037675） 作者单位:410008 长沙，中南大学湘雅医院泌尿外科（祖雄兵、齐琳、张向阳）