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目的:拟探讨RegⅣ基因在胃癌细胞中的表达及干扰该基因对胃癌细胞增殖和凋亡的影响.方法:应用实时定量PCR方法检测九株胃癌细胞株中RegⅣ基因的mRNA表达.针对RegⅣ基因设计三条小干扰RNA片段(siRNA1、siRNA2、siRNA3),瞬时转染高表达胃癌细胞株,实时定量PCR方法检测转染后RegⅣ基因的mRNA的表达水平,CCK-8法检测细胞增殖能力,流式细胞仪检测细胞凋亡.结果:以永生化正常胃黏膜细胞株GES-1作为参照,除MKN-45和SNU-1胃癌细胞株外,其余胃癌细胞株中RegⅣ表达水平均升高5倍以上,尤以SNU-16细胞株明显,其RegⅣ的表达水平高出GES-1细胞数千倍,遂选用SNU-16细胞进行RNA干扰.合成的三对siRNA对RegⅣ基因表达均有明显抑制作用,抑制率分别为79.3%、77.4%和60.4%.选用siRNA1干扰SNU-16细胞株后96h和120h,其细胞增殖率受到明显抑制(P=0.0057,0.0173).流式细胞仪检测显示72h细胞凋亡率明显增加(P=0.0231).结论:干扰RegⅣ基因具有抑制胃癌细胞增殖,促进凋亡的作用,RegⅣ基因可能成为胃癌基因靶向治疗的分子靶点.
Objective: To investigate the expression of RegⅣ gene in gastric cancer cells and to interfere with the effect of this gene on the proliferation and apoptosis of gastric cancer cells.Methods: Real-time quantitative PCR was used to detect the mRNA expression of RegⅣ gene in nine gastric cancer cell lines. Three small interfering RNA fragments (siRNA1, siRNA2 and siRNA3) were transiently transfected into gastric cancer cell lines. The expression of RegⅣ mRNA was detected by real-time PCR. The proliferation of cells was detected by CCK-8, The apoptosis of gastric cancer cells was detected by flow cytometry.Results: The expression levels of RegⅣ in gastric cancer cell lines except MKN-45 and SNU-1 gastric cancer cell lines were all increased by more than 5 times with GES-1 immortalized normal gastric mucosal cell line as reference, Especially in SNU-16 cell line, the expression level of RegⅣ was several thousand times higher than that of GES-1 cells, SNU-16 cells were selected for RNA interference.The three pairs of siRNAs were significantly inhibited RegⅣ gene expression, inhibition The rates of proliferation were significantly inhibited at the rates of 79.3%, 77.4% and 60.4%, respectively. The proliferation of SNU-16 cells was significantly inhibited at 96h and 120h (P = 0.0057,0.0173). Flow cytometry showed apoptosis at 72h The rate was significantly increased (P = 0.023 1) .Conclusion: The interference of RegⅣ gene can inhibit the proliferation and promote the apoptosis of gastric cancer cells, and RegⅣ gene may be the molecular target of targeted therapy of gastric cancer.