shRNA沉寂HK-Ⅱ表达对胃癌SGC7901细胞增殖、凋亡的影响

shRNA-mediated silencing of hexokinase Ⅱ inhibits proliferation but promotes apoptosis in SGC7901 cells

目的:观察shRNA沉寂HK-Ⅱ基因的表达对胃癌细胞株SGC7901细胞增殖、凋亡的影响,初步评价HK-Ⅱ基因治疗胃癌的应用前景.方法:取胃癌细胞株SGC7901及胃上皮细胞株GES-1细胞为研究对象,将每株细胞分别分成5组(A:干扰组;B:阳性对照组;C:阴性对照组;D:脂质体组;E:空白对照组).用shRNA沉寂胃癌细胞株SGC7901及胃上皮细胞株GES-1细胞中HK-Ⅱ的表达.用逆转录-聚合酶链反应(RT-PCR)检测转染shRNA后每株细胞中各组HK-ⅡmRNA的表达变化;分别用MTT及流式细胞仪检测转染后细胞增殖、凋亡的变化.结果:HK-ⅡmRNA在胃癌细胞株SGC7901细胞中的表达明显高于胃上皮细胞株GES-1细胞(t=12.119,P<0.01),HK-ⅡshRNA可以明显抑制两株细胞中HK-Ⅱ的表达(P<0.01).沉寂HK-Ⅱ的表达可抑制SGC7901细胞的增殖(F=159.811,P<0.01),并促进其凋亡(χ2=21.324,P<0.01);但对胃上皮细胞增殖(F=0.704,P=0.592)及凋亡(χ2=1.007,P=0.909)无明显影响.结论:沉寂HK-Ⅱ表达可以抑制胃癌细胞株SGC7901的增殖,促进其凋亡;但对胃上皮细胞株GES-1无明显影响.HK-Ⅱ可能成为胃癌治疗的一个靶点.

AIM：To observe the changes in cell proliferation and apoptosis in gastric cancer cell line SGC7901 after silencing the hexokinase II （HK-II） gene with an HK-II-specific shRNA, and to assess the potential application of HK-II-targeted gene therapy for gastric cancer.METHODS：Gastric cancer cell line SGC7901 and gastric epithelial cell line GES-1 were used in this study.Both SGC7901 cells and GES-1 cells were divided into 5 groups：HK-II shRNA group, positive control group, negative control group, empty liposome group, and blank control group.After transfection of HK-II-specific shRNA into SGC7901 and GES-1 cells, the change in the expression of HK-II mRNA was detected by reverse transcription-polymerase chain reaction （RT-PCR）, and the changes in cell proliferation and apoptosis were assessed by MTT assay and flow cytometry, respectively.RESULTS：The expression level of HK-II mRNA in SGC7901 cells was significantly higher than that in GES-1 cells （t=12.119, P0.01）.The expression of HK-II mRNA was obviously silenced after transfection of HK-II-specific shRNA （P0.01）.HK-II knockdown could significantly inhibit proliferation （F=159.811, P0.01） and promote apoptosis （χ2=21.324, P0.01） in SGC7901 cells, but had no significant effect on proliferation and apoptosis in GES-1 cells （F=0.704, P=0.592; χ2=1.007, P=0.909）.CONCLUSION：HK-II knockdown significantly inhibits proliferation and promotes apoptosis in SGC7901 cells, but has no significant impact in GES-1 cells.HK-II may be a potential target for the treatment of gastric cancer.