RNA干扰沉默ERK2基因体外诱导HepG2细胞凋亡的实验研究

Experimental study of RNA interference induce apoptosis of HepG2 cells by silencing ERK2 gene in vitro

目的:探讨RNA干扰技术对肝癌HepG2细胞ERK2基因表达的抑制作用及对HepG2细胞凋亡的作用。方法:实验分siRNA干扰质粒转染组、阴性质粒对照组和HepG2细胞空白对照组。应用DNA重组技术,制备针对ERK2基因的重组质粒pAAV-ERK2-siRNA-GFP,通过脂质体转染HepG2细胞。MTT细胞增殖实验,TUNEL法细胞凋亡染色及流式细胞仪凋亡细胞法检测体外诱导HepG2细胞凋亡的作用。结果:成功构建ERK2-siRNA重组质粒。MTT细胞增殖实验,TUNEL法细胞凋亡染色及流式细胞仪凋亡细胞检测法显示该重组质粒在体外能够有效抑制HepG2细胞生长,诱导HepG2细胞凋亡。结论:构建ERK2-siRNA重组质粒能抑制HepG2细胞增殖并诱导HepG2细胞凋亡,为肝癌治疗提供新思路。

Objective ：To investigate the inhibitory effect of RNA interference technology on ERK2 gene expression and the induction effect of apoptosis of HepG2 cells. Methods：The experiment was divided into siRNA interference plasmid transfection group, negative plasmid control and HepG2 cells blank control group. The recombinant plasmids pAAV ERK2 -siRNA -GFP against the ERK2 gene were prepared by recombinant DNA technology, and transfected into HepG2 cells by liposomes. HepG2 cells apoptosis was assessed by MTT cell proliferation experiments, TUNEL as- say apoptosis staining and flow cytometry assay in vitro. Results： ERK2 - siRNA recombinant plasmids were success- fully constructed. All the experiments show that the ERK2 - siRNA recombinant plasmids can induce HepG2 cells ap- optosis in vitro. Conclusion ： ERK2 - siRNA recombinant plasmid can inhibit the proliferation of HepG2 cells and in- duce HepG2 cells apoptosis, and this method may be a promising therapeutic strategy for liver cancer.