Rap1GAP对胃癌细胞侵袭、迁移及上皮细胞-间充质转化的作用

The Effect of Rap1GAP on Cell Invasion,Migration and Epithelial-Mesenchymal Transition in Gastric Cancer Cells

[目的]分析Rap1GAP对胃癌细胞增殖、侵袭及迁移能力的影响。[方法] 运用qRT-PCR及Western blo法检测Rap1GAP在胃癌细胞中的表达。Rap1GAP载体慢病毒感染MGC803及MKN-45两个细胞系,构建出Rap1GAP高表达胃癌细胞系。MTT、细胞克隆形成实验、Transwell实验及划痕实验检测Rap1GAP 对胃癌细胞增殖、侵袭及迁移能力的影响。运用qRT-PCR及Western blot法检测上皮化标志物E-cadherin、Snail和N-cadherin的表达。[结果]正常细胞株GES-1中Rap1GAP表达量为1.02±0.08,而SGC7901、AGS、MGC803、HGC-27、MKN-45中表达量分别为0.66±0.10、0.51±0.10、0.20±0.08、0.31±0.07、0.26±0.11(P均<0.01)。体外实验显示Rap1GAP能抑制胃癌细胞的增殖(酶标仪检测波长为570 nm时的吸光度)、侵袭及迁移能力;且Rap1GAP能抑制胃癌细胞的EMT。[结论] Rap1GAP在胃癌进展中起到抑癌作用,具有成为胃癌治疗靶点的潜力。

[Objective]To investigate the effect of Rap1GAP on the proliferation,invasion and migration of gastric cancer cells. [Methods] The expression of Rap1GAP in gastric cancer cells was determined by qRT-PCR and Western blot. Gastric cancer MGC803 and MKN-45 cells with high expression of Rap1GAP were constructed by infection with Rap1GAP lent virus vector. The effect of Rap1GAP on the cell proliferation was detected by MTT assay and cell clone formation assay. The effect of Rap1GAP on the cell invasion was detected by Transwell assay. The effect of Rap1GAP on the cell migration was detected by Scratch assay. The expression of epithelial markers (E-cadherin,Snail and N-cadherin) was detected by qRT-PCR and Western blot. [Results]The expression level of Rap1GAP in normal gastric GES-1 cells was 1.02±0.08,while the expression level in SGC7901,AGS,MGC803,HGC-27 and MKN-45 cells was 0.66±0.10,0.51±0.10,0.20±0.08,0.31±0.07 and 0.26±0.11,respectively (all P<0.01). Rap1GAP can inhibited the proliferation,invasion,migration and EMT of gastric cancer cells. [Conclusion] Rap1GAP can inhibit proliferation,migration and invasion of gastric cancer cell,which may be potentially used as a therapeutic target for gastric cancer.