咪达唑仑通过下调circASXL1抑制胃癌细胞增殖、迁移和侵袭

Midazolam inhibits the proliferation,migration and invasion of gastric cancer cells by down-regulating circASxL1

目的探讨咪达唑仑对胃癌细胞增殖、迁移和侵袭的可能机制。方法体外培养胃癌HGC-27细胞,用不同剂量(10、20、40μmol/L)咪达唑仑干预24 h后,CCK-8法、克隆形成实验、Transwell分别检测细胞增殖抑制率、克隆数、迁移及侵袭,蛋白质印迹法检测细胞中E-cadherin和N-cadherin蛋白表达,qRT-PCR法检测细胞中circASXL1表达。分别以53例癌旁组织、胃黏膜上皮细胞GES-1为对照,qRT-PCR法检测53例胃癌组织和HGC-27细胞中circASXL1表达。同时,咪达唑仑干预HGC-27细胞24 h后,qRT-PCR法检测细胞中circASXL1表达。转染circASXL1小干扰RNA或过表达载体至HGC-27细胞,上述相同方法观察circASXL1对HGC-27细胞增殖、迁移和侵袭的影响。结果与对照组比较,HGC-27细胞经不同剂量咪达唑仑干预后,细胞增殖抑制率、E-cadherin蛋白表达均升高(P<0.05),细胞克隆数、迁移数、侵袭数及N-cadherin蛋白表达均降低(P<0.05)。胃癌组织中circASXL1的表达明显高于癌旁组织(P<0.05);胃癌HGC-27细胞中circASXL1表达明显高于GES-1细胞(P<0.05)。敲减circASXL1后,HGC-27细胞增殖抑制率、E-cadherin蛋白表达均升高(P<0.05),细胞克隆数、迁移数、侵袭数及N-cadherin蛋白表达均降低(P<0.05)。与对照组比较,不同剂量咪达唑仑降低了HGC-27细胞中circASXL1的表达,过表达circASXL1降低了咪达唑仑对HGC-27细胞增殖、迁移和侵袭的影响。结论咪达唑仑可能通过下调circASXL1抑制胃癌HGC-27细胞增殖、迁移和侵袭。

Objective To investigate the effect of midazolam on the proliferation,migration and invasion of gastric cancer cells and its possible mechanism.Methods HGC-27 cells of gastric cancer were cultured in vitro and treated with different doses(10,20,40μmol/L)of midazolam for 24 h,and then CCK-8 assay,clone formation assay,and Transwell were used to detect cell proliferation inhibition rate,clone number,migration and invasion,respectively.The protein expression of E-cadherin and N-cadherins in the cells were detected by Western blotting.The expression of circASXL1 in the cells was detected by qRT-PCR.With 53 cases of adjacent tissues and gastric mucosal epithelial cells GES-1 as controls,qRT-PCR method was used to detect the expression of circASXL1 in 53 cases of gastric cancer tissues and HGC-27 cells.At the same time,the expression of circASXL1 in HGC-27 cells that interfered by midazolam for 24 hours was detected by qRT-PCR.circASXL1 small interfering RNA or overexpression vector was transfected into HGC-27 cells,and the same methods as above were used to observe the effect of circASXL1 on the proliferation,migration and invasion of HGC-27 cells.Results Compared with the control group,after HGC-27 cells were interfered with different doses of midazolam,the cell proliferation inhibition rate and the protein expression of E-cadherin were increased(P<0.05),but the number of cell clone,migration and invasion,and the protein expression of N-cadherin were decreased(P<0.05).The expression of circASXL1 in gastric cancer tissue was significantly higher than that in adjacent tissues(P<0.05);the expression of circASXL1 in gastric cancer HGC-27 cells was significantly higher than that in GES-1 cells(P<o.05).After knocking down circASXL1,the cell proliferation inhibition rate and the protein expression of E-cadherin were increased(P<o.05),but the number of cell clone,migration and invasion,and the protein expression of N-cadherin were decreased(P<o.05).Compared with the control group,different doses of midazolam reduced the expression of circASXL1 in HGC-27 cells,and overexpression of circASXL1 reduced the effects of midazolam on the proliferation,migration and invasion of HGC-27 cells.Conclusion Midazolam may inhibit the proliferation,migration and invasion of gastric cancer HGC-27 cells by down-regulating circASXL1.