应用RNA干扰技术研究PPARδ基因对人大肠癌细胞增殖能力的影响

RNA Interference against Peroxisome Proliferator-Activated Receptor δ Gene Promotes Proliferation of Human Colorectal Cancer Cells

目的:探索PPARδ基因对大肠癌细胞增殖能力的影响。方法:构建表达短发夹结构RNA(short-hair-pinRNA,shRNA)的质粒载体,采用脂质体2000将质粒载体转染入大肠癌细胞株HCT-116,应用实时荧光定量逆转录PCR分析PPARδ基因沉默效果,采用噻唑蓝(MTT)比色实验、及流式细胞术观察HCT-116细胞株增殖及细胞周期的变化。结果:MTT试验显示,转染后24～96h,干扰组细胞在各时点的光吸收值均显著高于对照组(P<0.05)。转染后48h,干扰组的G1期细胞分布比对照组显著降低(26.8%VS38.6%,P=0.037),各组在S期与G2/M期的细胞分布比例(S期:G2/M期)无显著差异(P=0.782)。结论:PPARδ基因可增加G1期的大肠癌细胞分布,并抑制细胞增殖。

Objective:To investigate the effect of peroxisome proliferator-activated receptor δ(PPARδ)on the proliferation of human colorectal cancer(CRC)cells.Methods:For RNA interfering(RNAi),HCT-116 cells were transfected with short hairpin RNA(shRNA)-expressing plasmids against PPARδ or negative control vectors.The efficacy of RNAi was assessed by real-time reverse transcription polymerase chain reaction(RT-PCR).The proliferation and the cell cycle of HCT-116 cells treated by RNAi were analyzed respectively using M...