siRNA沉默HIF-1α基因对前列腺癌PC-3细胞多西紫杉醇化疗敏感性的影响  

Knocking down the HIF-1α gene with siRNA increases the chemosensitivity of prostate cancer PC-3 cells to docetaxel

作　　者：付振宇 张鸽[1,2] 黄玉华 严春寅[3] 俞江[4] 孙利国[1,2] 陈永昌[1,2] 张杰 

机构地区：[1]扬州大学第五临床学院 [2]常熟市第二人民医院泌尿外科,江苏常熟215500 [3]苏州大学附属第一医院泌尿外科,江苏苏州215006 [4]上海交通大学医学院附属苏州九龙医院泌尿外科,江苏苏州215000

出　　处：《中华男科学杂志》2014年第10期881-885,共5页National Journal of Andrology

摘　　要：目的:探讨沉默缺氧诱导因子-1α(HIF-1α)基因对前列腺癌PC-3细胞多西紫杉醇(docetaxel,DTX)化疗敏感性的影响。方法:实验分为PC-3组,PC-3+NCsiRNA组,PC-3+HIF-lαsiRNA组。用脂质体Lipofectamine2000将经过筛选证实有效的HIF-1αsiRNA序列和阴性NC siRNA序列转染PC-3细胞。待细胞转染或不转染后继续培养,根据实验要求时间点加入DTX。采用CCK-8法检测转染后96 h内各组细胞的生长增殖情况,流式细胞仪检测加DTX后48 h各组细胞的周期分布、凋亡率及多药耐药基因P糖蛋白(MDR/P-gp)的表达。结果:PC-3+HIF-1αsiRNA组肿瘤抑制率高于PC-3+NC siRNA组,尤以转染后48 h表现的较为明显;转染后48 h,PC-3+HIF-1αsiRNA组细胞存活率均明显低于PC-3组和PC-3+NCsiRNA组(P均<0.01)。细胞周期分布变化,PC-3+HIF-1αsiRNA组较两对照组比较,G0/G1期细胞百分数较低,而G2/M期细胞百分数稍高(P均<0.01)。细胞凋亡结果,PC-3+HIF-1αsiRNA组细胞凋亡率明显高于两对照组(P均<0.01)。各组细胞MDR/P-gp表达无明显差异(P均>0.05)。结论:沉默HIF-1α基因能增强DTX对前列腺癌PC-3细胞的生长抑制,通过调整细胞周期重新分布、诱导细胞凋亡,增加前列腺癌PC-3细胞对化疗的敏感性,而这一作用与MDR/P-gp无明显相关。Objective： To explore the effect of knockdown of the HIF-1α gene on the chemosensitivity of PC-3 cells to docetaxel（ DTX）. Methods： PC-3 cells were divided into groups A（ PC-3）,B（ PC-3 ＋ NC siRNA）,and C（ PC-3 ＋ HIF-1α siRNA）.Using cationic liposomes Lipofectamine 2000,the cells in groups B and C were transfected with negative NC siRNA and HIF-1α-siRNA,respectively,which had been confirmed to be effective in the previous study. After transfection,all the cells were cultured in the medium with DTX at the concentration of 0. 25 μmol /L. Then the proliferation of the PC-3 cells within 96 hours was measured via the CCK-8 test. The cycle distribution and apoptosis rate of the PC-3 cells and the expression of the multidrug-resistance gene /P glucose protein（ MDR /P-gp） were determined by flow cytometry at 48 hours after DTX treatment. Results： The survival rate of the PC-3 cells was significantly lower in group C than in A and B,particularly at 48 hours after transfection（ P〈 0. 01）. Flow cytometry analysis showed a lower percentage of the PC-3 cells in the G0 /G1 phase and a little higher percentage in the G2 /M phase in group C than in the other two groups（ P〈 0. 01）. The rate of cell apoptosis was remarkably higher in group C than in A and B（ P 〈0. 01）. There was no significant difference in the expression of MDR /P-gp among the three groups（ P〉 0. 05）. Conclusion： Knocking down the HIF-1α gene expression may enhance the inhibitory effect of DTX on PC-3 cells by regulating the distribution of cell cycles,inducing cell apoptosis,and increasing the chemosensitivity of the cells to DTX,but with no obvious association with the expression of MDR /P-gp.

关 键 词：PC-3细胞 缺氧诱导因子-1α 增殖 细胞凋亡 细胞周期 化疗增敏 前列腺癌 

分 类 号：R737.25[医药卫生—肿瘤]