靶向sKDR对膀胱癌T24细胞增殖和凋亡的影响

Effects of target sKDR on proliferation and apoptosis of bladder cancer T24 cells

目的:观察靶向可溶性血管内皮生长因子受体(sKDR)对膀胱癌细胞增殖和凋亡的影响,探讨KDR基因作为膀胱癌基因治疗靶点的可行性和有效性。方法:将人膀胱癌细胞T24分为sKDR转染组、未转染组和对照组,构建靶向sKDR真核分泌型表达质粒PCI-KDR,经LipofctamineTM2000转染T24细胞,采用酶联免疫吸附实验(ELISA)检测能否与VEGF结合,逆转录聚合酶链反应(RT-PCR)检测稳定表达sKDR的T24细胞KDR基因表达水平,噻唑蓝比色法(MTT)检测sKDR对T24细胞增殖的作用,转移酶末端标记法(TUNEL)检测细胞凋亡率,裸鼠皮下移植稳定表达sKDR的T24细胞,观察其成瘤性的改变。结果:与未转染组比较,转染组稳定表达sKDR的T24细胞其KDRmRNA表达水平升高了4.16倍,细胞增殖抑制率增加了55.1%(P<0.01),细胞增殖速度明显缓慢并出现显著的凋亡峰。在裸鼠体内转染组与未转染组上述各项指标比较差异有显著性(P<0.01),转染组肿瘤生长明显缓慢,成瘤能力受抑。结论:靶向sKDR能够在很大程度上抑制膀胱癌T24细胞增殖并促进其凋亡。

Objective To observe the effects of target solubility kinase insert domain receptor （sKDR） on proliferation and apoptosis of human bladder cancer T24 cells and the feasibility and effectiveness of sKDR gene as the target of bladder cancer gene therapy . Methods Human bladder cancer T24 cells were divided into sKDR transfected group, untransfected group and contol group. Eukaryotic secretory expression plasmid PCl-sKDR was constructed and T24 cells were transfected by Lipofectamine^TM 2000 and further screened by ELISA . The expressions of sKDR gene in at all groups were determined by RT-PCR, the proliferation of T24 cells in these groups was measured by MTT method, the apoptotic rates were detected by TUNEL. T24 cells treated with sKDR were transplanted subcutanuously in nude mice and the tumorgenesis ability was observed. Results The expression of KDR mRNA in the KDR- expression T24 cells were 4.16 times that in vacant vector PCk-neo cells. Compared with PCl-neo contol, the proliferatory inhibitory rate of cells with sKDR- expression was increased by 55.1 % （P〈 0.01）. The tumorgenesis in transfected group was significantly restrained compared with untransfected group （P〈 0.01）. Conclusion sKDR can inhibit the proliferateion and promote the apoptosis of bladder cancer T24 cells to a certain degree.