RNA干扰Hedgehog信号通路对胃癌细胞增殖和凋亡影响及其机制的探讨

目的:观察RNA干扰Hedgehog信号通路关键因子Glil对胃癌细胞的生物学行为的影响并分析相关机制.方法:人工合成靶向Hedgehog信号通路活化关键因子Glil的siRNA(Glil siRNA)和对照无关siRNA(Con siRNA),应用脂质体将其转入人胃癌细胞SGC-7901,48 h后收集细胞,分别应用MTT法和流式细胞仪检测对细胞增殖、周期和凋亡的影响,同时应用TaqMan探针实时定量PCR检测靶基因Glil、Hedgehog活化标志基因PTCH1、细胞周期负调控蛋白基因CDKN1A(p21)和抗凋亡基因Bel-2的表达变化.结果:以单纯转染剂对照组为参照,siRNA转染后48 h,Glil siRNA和Con siRNA的抑制率分别为(53.33±6.06)%和(13.33±6.11)%,两组差异有统计学意义,t=5.701,P=0.005,n=3;G0/G1期分别为(80.67±4.51)%和(66.00±3.10)%,两组差异有统计学意义,t=5.500,P=0.005,n=3;凋亡率分别为(15.97±1.76)%和(7.77±1.11)%,两组差异有统计学意义,t=4.671,P=0.01,n=3;GlilsiRNA转染细胞内Glil、PTCH1、Bcl-2和CDKN1A(p21)基因的相对(参照单纯转染剂)表达分别为对照的0.24±0.11、0.43±0.09、0.52±0.13和2.10±0.30.分别与Con siRNA相比4条基因表达均差异有统计学意义,t=9.759,P=0.001,n=3;t=8.645,P=0.001,n=3;t=4.940,P=0.008,n=3;t=5.962,P=0.004,n=3.结论:应用RNA干扰技术可以阻断Hedgehog信号通路关键因子Glil的表达,抑制Hedgehog信号通路活化,从而有效地抑制胃癌细胞的增长,诱导凋亡,可能成为治疗胃癌新的生物靶向治疗途径.

Effect of Hedgehog signaling pathway interfered by siRNA in gastric cancer cell proliferation and apoptosis and its mechanism

OBJECTIVE:To study the effect of the small interfering RNA targeting Gli1(Gli1 siRNA),and the key factor of Hedgehog signaling pathway,on the biological behavior of gastric cancer cells and its mechanism.METHODS: Synthetic Gli1 siRNA and the irrelevant control siRNA(Con siRNA) were transferred into gastric cancer SGC-7901 cells.After 48 h the cell proliferation and the cell cycle and apoptosis were detected by MTT and flow cytometry.Simultaneously,the mRNA expression of the relevant genes including Gli1,PTC...