靶向EGFR基因RNA干扰对胃癌细胞BGC823细胞增殖和凋亡的影响

目的利用R NA干扰技术下调表皮生长因子受体(EGFR)基因的表达,观察其对胃癌BGC823细胞增殖和凋亡的影响。方法将细胞分为3组,空白对照组、空载体组和实验组(转染EGFRshRNA组)。构建针对EGFR基因的s和R NA真核表达载体,转染入胃癌BGC823细胞中,运用R T-PCR和Western blot检测EGFR在mRNA和蛋白水平的变化;WST-1法检测细胞增殖的影响:Hoechst33258染色观察细胞核形态学变化。实验重复3次。结果成功构建表达质粒pGenSil-EGFR并转染BGC823细胞,shRNA下调EGFR的表达后,与空白对照组相比EGFR在mR NA和蛋白水平的表达均下降,抑制率分别为58.6%和75.2%。shR NA转染24、48、72 h,WST-1法检测胃癌细胞BGC823的增殖情况,与空白对照组相比,24 h后三组差异不明显(F=0.326,P>0.05),48 h及72 h后,细胞增殖明显下降(F=68.25及274.45,P<0.001),有统计学意义。EGFR shR NA作用BGC823细胞72 h后,经Hoechst33258染色发现,实验组细胞核染色质边集,出现凋亡小体。结论靶向EGFR基因RNA干扰能够阻抑胃癌细胞BGC823细胞的EGFR表达,诱导细胞凋亡,抑制细胞增殖,EGFR基因有望成为胃癌基因治疗靶点。

RNA interference targeting EGFR on proliferation and apoptosis of human gastric carcinoma cell line BGC823

【Objective】 To explore the effect of down-regulating epidermal growth factor receptor(EGFR) expression by RNA interference on the proliferation of gastric carcinoma BGC823 cells.【Methods】 BGC823 cells were divided into three groups: blank control group,negative control group(transfected with an empty vector),and experiment group(transfected with EGFR shRNA).The recombinant eukaryotic expression plasmids pGenSil-EGFR were constructed and transfected into BGC823 cells respectively,the mRNA and protein expression of EGFR was examined by RT-PCR and Western blot,respectively;cell proliferation was evaluated by WST-1assay;and cell apoptosis was detected by Hoechst33258.【Results】 The expressions of EGFR in BGC823 cells transfected with pGenSil-EGFR were inhibited significantly at both mRNA and protein levels,with an inhibitory rate of 58.6%and 75.2%.At 48,and 72 h after transfection,the proliferation rates of BGC823 cells transfected with EGFR shRNA significantly lower than those of non-transfected cells(F =68.25 and 274.45,P <0.001),24 h besides(F =0.326,P >0.05).After 72 h,the BGC823 transfected with EGFR shRNA was proved to be more apoptosis than those of cells of the blank control group and negative control by Hoechst33258 dyeing.【Conclusion】 RNA interference targeting EGFR gene down-regulates the EGFR expression,induces cell apoptosis and inhibits cell proliferation in gastric carcinoma BGC823 cells.EGFR may be a new gene therapy target for gastric cancer.