Objective: To investigate the role of ERK in the apoptosis of gastric cancer cells induced by miR-433. Methods: Lentivirus was used to transfect BGC-823 gastric cancer cell line to over-express miR-433. The blank control group (BGC-823), negative control group (BGC-823+ miR-433 negative control) and experimental group (miR-433+ miR-433, BGC-823-pMD18-T- miR-433)were set up. After treatment, the gastric cancer cell line BGC-823 was detected at 24 h, 48 h, and 72 h after culture, in vitro cell activity, cell apoptosis assays were performed by CCK-8 and Annexinv-FITC, respectively, to elucidate biological effects of miRNA-433, and After 72 h of culture, the ERK1/2 detected their protein expression were quantified by BCA method. Result: The growth activity of BGC-823+ miR-433 cells cultured in vitro was significantly lower than that of BGC-823 cells and BGC-823+ miR-433 negative control cells at 48 h and 72 h; BGC-823+ miR-433 cell apoptosis index was significantly increased at 24 h, 48 h, and 72 h; the expression of ERK1/2 was significantly lower than BGC-823 cells and BGC-823+ miR-433 negative contral after 72 h culture. There were no significant differences between BGC-823+ miR-433 negative control cells and BGC-823 cells. Conclusion: ERK plays an important role in the apoptosis of gastric cancer cells induced by miR-433.
目的： 探讨胞外信号调节激酶(ERK1/2)通路在miR－433诱导胃癌细胞凋亡中的作用。方法： 使用慢病毒转染胃癌BGC－823细胞株使其过表达miR－433，实验设置空白对照组(BGC－823)、阴性对照组(BGC－823＋ miR－433空载体)和实验组(miR－433＋ miR－433，BGC－823－pMD18－T－ miR－433)。经过相应处理后胃癌细胞株BGC－823分别于培养后24、48、72 h后使用cck－8试剂盒行细胞活性检测和Annexinv－FITC细胞凋亡实验，并于培养72 h后使用BCA法蛋白定量对ERK1/2检测其蛋白表达。结果： 体外培养的BGC－823＋miR－433细胞生长活性在48 h(0.387±0.005)和72 h(0.614±0.009)时间点上显著低于BGC－823细胞(0.544±0.009、1.008±0.012)和BGC－823＋miR－433阴性对照细胞(0.526±0.008、0.964±0.006)，差异均有统计学意义；BGC－823＋miR－433细胞凋亡指数在24、48 h和72 h时显著增加；ERK1/2的表达在BGC－823＋miR－433细胞培养72 h后显著低于BGC－823细胞组和BGC－823＋miR－433阴性对照组；在各个检测中BGC－823细胞和BGC－823＋miR－433阴性对照细胞差异均无统计学意义。结论： ERK1/2在miR－433诱导胃癌细胞凋亡中起重要作用。.
Keywords: ERK1/2 Pathway; Stomach neoplasms; miR-433.