胃癌细胞中RASSF6基因表达及对胃癌细胞增殖迁移的影响

Expression of RASSF6 Gene and Its Effect on Proliferation and Migration of Gastric Cancer Cells

目的探讨RASSF6对胃癌细胞增殖迁移和凋亡的影响。方法采用实时荧光定量聚合酶链反应(quantitative real-time polymerasechain reaction,qRT-PCR)、蛋白质印迹法(Western blot)检测胃癌细胞系BGC823、MGC803及正常胃黏膜上皮细胞系GES-1中RASSF6 mRNA及蛋白表达情况,筛选出低表达的BGC823细胞,通过慢病毒转染技术构建慢病毒对照组(negative control,NC)和RASSF6基因过表达组(over expression,OP),通过CCK-8法、细胞划痕检测各组细胞增殖、迁移能力,流式细胞技术检测各组细胞凋亡情况。结果RASSF6基因及蛋白在胃癌细胞系中表达量均低于正常胃黏膜上皮细胞系(P均<0.05)。CCK-8细胞增殖检测OP组细胞24 h、48 h的OD值均低于NC组,细胞划痕面积比值、细胞凋亡率高于NC组(P均<0.05)。结论RASSF6在胃癌细胞中低表达,上调RASSF6可抑制胃癌BGC823细胞增殖、迁移,促进凋亡,RASSF6可能为胃癌的抑癌基因。

Objective To investigate the effect of RASSF6 on proliferation,migration and apoptosis of gastric cancer cells.Methods q RT-PCR and Western blot were used to detect gastric cancer cell lines BGC823,MGC803 and normal gastric epithelial cell line GES-1’s RASSF6 m RNA and protein expression.The RASSF6 gene of BGC823 cells which was the lowest expression of RASSF6 m RNA was infected by lentivirus control group and RASSF6 gene overexpression lentivirus vector.CCK-8 method and wound healing assay were used to detect the changes of cell proliferation and invasion,cell apoptosis was detected by flow cytometry in the two groups.Results The level of RASSF6 gene and protein expression in human gastric cancer cell lines was lower than normal gastric mucosa epithelial cells(P all<0.05).OD value of OP group was lower than NC group at 24 h and 48 h.Cell scratch area ratio and cell apoptosis rate were significantly higher than NC group(P all<0.05).Conclusion RASSF6 is poorly expressed in gastric cancer cells,and up-regulation of RASSF6 can inhibit the proliferation,migration and promote apoptosis of gastric cancer BGC823 cells.It may be a tumor suppressor gene for gastric cancer.