缺氧诱导因子-1α小干扰RNA对人膀胱癌小鼠移植瘤生长的实验研究

目的 观察缺氧诱导因子-1α(HIF-1α)siRNA对人膀胱癌小鼠膀胱原位移植瘤生长的抑制作用,探讨HIF-1α作为膀胱癌基因治疗靶点的有效性.方法 把已稳定转染pGC-siHIF-1α的人膀胱癌细胞株T24接种至小鼠膀胱.通过CT观察肿瘤生长,应用免疫组化(SABC)检测肿瘤细胞HIF-1α和肿瘤间质CD34的水平,根据CD34表达情况评价肿瘤微血管密度(MVD).末端脱氧核苷酸转移酶标注法(TUNEL)测定肿瘤细胞凋亡指数;二苯基溴化四氮唑蓝(MTT)法测定肿瘤细胞增值率;流式细胞仪(FCM)测定肿瘤细胞周期分布.结果 实验组与对照组相比较,实验组成瘤率低,肿瘤生长速度缓慢(p＜0.05);HIF-1α水平下降(P＜0.01);肿瘤微血管密度减少(P＜0.01);凋亡指数升高(P＜0.01);增殖能力减弱(P＜0.01);G0/GI期细胞增多,G2/M期和S期细胞均减少(P＜0.05);与对照组相比较,组间差异均具有统计学意义.结论 以HIF-1α为靶点的siRNA有抑制H1F-1α表达,遏制膀胱癌T24细胞在体内生长的作用,HIF-1 0有可能成为临床膀胱癌基因治疗的新的有效靶点.

The experimental research of hypoxia-inducible factor-1α small interfering RNA on the growth of human bladder cancer xenografts in mice

Objective To observe the inhibitory effect of hypoxia inducible factor-1α(HIF-1α)siRNA on the growth of human bladder cancer xenografts in mice,and to explore the effectiveness of HIF-1α as a target for bladder cancer gene therapy.Methods pGC-siHIF-1α of stably transfected into human bladder cancer cell line T24 was inoculated to mice bladder,and the mouse model of bladder cancer was established.The tumor growth was observed by CT,and the level of tumor cells HIF-1α and CD34 was detected by immunohistochemistry (SABC),and the expression of tumor microvessel density (MVD) was evaluated according to the expression of CD34.The apoptosis index of tumor cells was determined by terminal transferase labeling (TUNEL).Results Comparison between experimental group and control group,compose got the lower rate experiment,slow tumor growth (P ＜0.05).HIF-1oα levels decreased (P ＜ 0.01),tumor microvessel density decreased (P ＜0.01),and the apoptosis index was significantly increased (P ＜0.01).Compared with the control group,the differences between the two groups were statistically significant.Conclusions As the target of HIF-1α,the siRNA have inhibited HIF-1ααexpression,contain T24 bladder cancer cells in vivo.HIF-lα may become a potential target for clinical gene therapy for bladder cancer,and provide experimental basis for anti-angiogenesis therapy of bladder cancer.