RNA干扰REG1A基因抑制人膀胱癌T24细胞增殖

Small interfering RNA targeting REG1A gene inhibits the proliferation of human bladder cancer cell line T24 in vitro

目的:观察小干扰RNA(small interfering RNA,siRNA)沉默REG1A(regenerating islet-derived1 alpha)基因的表达对人膀胱癌T24细胞增殖及细胞周期的影响。方法:化学合成针对REG1A基因的3对siRNA,使用脂质体将其转染至T24细胞中。应用实时荧光定量PCR和蛋白质印迹法分别检测REG1AsiRNA转染后T24细胞REG1AmRNA和蛋白的表达,CCK-8(cell counting kit-8)法检测转染后的细胞增殖情况,FCM检测转染后细胞周期的变化。结果:与空白对照组比较,REG1AsiRNA转染T24细胞后,REG1AmRNA和蛋白表达分别下降了(83.10±0.06)%和(55.81±0.16)%,差异有统计学意义(P<0.05);细胞增殖受到抑制(P<0.05),G0/G1期细胞百分比明显增加(P<0.05),细胞增殖指数下降(P<0.05)。结论:REG1AsiRNA能有效抑制膀胱癌T24细胞REG1AmRNA和蛋白的表达,并抑制细胞增殖。

Objective：To investigate the effects of silencing regenerating islet-derived 1 alpha（REG1A） gene by small interfering RNA（siRNA） on the cell proliferation and the cell cycle of the human bladder cancer cell line T24.Methods：Three siRNAs targeting human REG1A were synthesized,which were transfected into T24 cells using a liposome approach.The expressions of REG1A mRNA and protein in T24 cells after transfection with REG1A siRNA were determined by real-time fluorescence quantitative-PCR（RFQ-PCR） and Western blotting,respectively.The cell proliferation and cell cycle were detected by cell counting kit-8（CCK-8） method and flow cytometry（FCM）,respectively.Results：As compared with the blank control（untransfected with siRNA）,the expression levels of REG1A mRNA and protein were decreased（83.10±0.06）% and（55.81±0.16）% after transfection with REG1A siRNA,respectively（P0.05）.The cell proliferation was inhibited（P0.05）,the cell cycle was arrested at G0/G1 phase（P0.05）,and the proliferation index was decreased（P0.05）.Conclusion：REG1A siRNA can effectively suppress the expressions of REG1A mRNA and protein in bladder cancer cell line T24,and inhibit the cell proliferation in vitro.