微小 RNA-588靶向调控脾酪氨酸激酶的表达及其对胃癌细胞增殖、凋亡的作用

目的 探讨微小RNA-588(miR-588)对胃癌细胞增殖、凋亡的影响以及潜在的作用机制.方法 运用实时荧光定量逆转录聚合酶链式反应(qRT-PCR)检测胃癌细胞SGC-7901、BGC-823和正常胃黏膜细胞GES-1中miR-588的表达水平;将胃癌细胞SGC-7901、BGC-823分为anti-miR-NC组、anti-miR-588组、anti-miR-588+si-NC组、anti-miR-588+si-脾酪氨酸激酶(Syk)组;蛋白质印迹法(Western blotting)检测蛋白表达;四甲基偶氮唑盐微量酶反应比色(MTT)法检测BGC-823细胞活性;流式细胞术检测BGC-823细胞凋亡率;双荧光素酶报告实验检测miR-588和Syk的靶向关系.结果 胃癌细胞SGC-7901、BGC-823中miR-588的表达水平高于GES-1细胞(0.58±0.06)、(0.84±0.08)比(0.36±0.04),P<0.05].抑制miR-588表达可抑制胃癌细胞增殖,诱导细胞凋亡;促进P21、Bax蛋白的表达,抑制cyclin D1、Bcl-2蛋白的表达.miR-588靶向负调控Syk的表达,抑制Syk表达能逆转抑制miR-588对细胞BGC-823细胞的作用.结论 miR-588可抑制胃癌细胞细胞增殖,诱导细胞凋亡,其机制可能与Syk有关.

Mir-588 targeted regulation of spleen tyrosine kinase expression and its effect on proliferation and apoptosis of gastric cancer cells

Objective To investigate the effect of miR-588 on proliferation and apoptosis of gastric cancer cells and its potential mechanism.Methods qRT-PCR was used to detect the expression level of miR-588 in gastric cancer cells SGC-7901, BGC-823 and normal gastric mucosal cells GES-1; gastric cancer cells SGC-7901 and BGC-823 were divided into anti-miR-NC group, anti-miR-588 group, anti-miR-588+si-NC group, anti-miR-588+si-Syk group; Western Blot was used to detect protein expression; MTT method was employed to detect BGC-823 cell viability; flow cytometry was performed to detect BGC-823 cell apoptosis rate; the dual luciferase re. porter experiment was used to detect the targeting relationship between miR-588 and Syk.Results The expression level of miR-588 in gastric cancer cells SGC-7901 and BGC-823 was higher than that of GES-1 cells (0.58±0.06),(0.84±0.08)vs.(0.36±0.04),P < 0.05]. In. hibition of miR-588 expression could inhibit the proliferation of gastric cancer cells, induce apoptosis, promote the expression of P21and Bax proteins, and inhibit the expression of cyclin D1 and Bcl-2 proteins. miR-588 targeting negatively regulated the expression of Syk, and inhibition of Syk expression could reverse the effect of inhibiting miR-588 on BGC-823 cells.Conclusion MiR-588 can in. hibit cell proliferation and induce apoptosis of gastric cancer cells, and its mechanism may be related to Syk.