摘要
目的:探讨miR-124对胃癌细胞增殖、凋亡与迁移的影响及其相关调控机制。方法:收集确诊为胃癌的癌组织和癌旁组织42组,采用实时定量PCR(qRT-PCR)检测癌和癌旁组织以及正常胃黏膜上皮细胞GES-1和胃癌细胞株BGC-823、MGC-803、SGC-7901、AGS中miR-124的相对表达量;在胃癌细胞株BGC-823中过表达miR-124,CCK-8法检测细胞增殖,流式细胞技术检测细胞凋亡,Transwell检测细胞迁移,Westernblot检测过表达miR-124前后磷酸化PI3K/AKT的变化。结果:胃癌组织中miR-124水平低于癌旁组织(0.70±0.15vs.1.14±0.27,t=10.14,P=0.000),胃癌细胞株中miR-124水平低于GES-1(F=38.012,P=0.000);过表达miR-124后细胞BGC-823的增殖与迁移能力均减低(0.94±0.09vs.0.62±0.06,t=6.615,P=0.000;152.33±14.05vs.53.67±12.58,t=9.061,P=0.001),而细胞凋亡率增加(2.10±0.56vs.8.30±0.31,t=16.777,P=0.000);过表达miR-124后p-AKT、p-PI3K蛋白表达均出现下调(118.90±10.51vs.34.87±4.23,t=12.847,P=0.000;158.53±10.94vs.71.07±4.51,t=12.802,P=0.000)。结论:miR-124在胃癌中低表达;miR-124抑制胃癌细胞的增殖、迁移,促进胃癌细胞凋亡,可能是靶向PI3K/AKT信号通路实现的。
Objective :To investigate the effects of miR-124 on the proliferation,apoptosis and migration of gastric cancer cells and its related regulatory mechanisms. Methods: Cancer and paracancerous tissues were obtained from 42 patients diagnosed as gastric cancer. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-124 in GES-1 and BGC-823,MGC-803,SGC-7901 and AGS in cancer and adjacent tissues as well as normal gastric mucosal epithelial cells besides overexpression of miR-124 in gastric cancer cell line BGC-823. CCK-8 assay was performed to determine the cell proliferation. Flow cytometry was used to detect the apoptosis,and Transwell assay for cell migration. Western-blot was performed to examine the changes of phosphorylated PI3K/AKT before and after overexpression of miR-124. Results: The level of miR-124 in gastric cancer tissues was significantly lower than that in adjacent tissues (0.70±0.15 vs. 1.14±0.27, t= 10.14, P= 0.000),and miR-124 level in gastric cancer cell lines was lower than that of GES-1 ( F= 38.012, P= 0.000). Proliferation and migration ability of BGC-823 cells were significantly decreased after overexpression of miR-124 (0.94±0.09 vs. 0.62±0.06, t= 6.615, P= 0.000;152.33±14.05 vs. 53.67±12.58, t= 9.061, P= 0.001),yet the apoptotic rate was increased (2.10±0.56 vs. 8.30±0.31, t= 16.777, P= 0.000). Protein p-AKT and p-PI3K were down-regulated after overexpression of miR-124 (118.90 ±10.51 vs. 34.87±4.23, t= 12.847, P= 0.000;158.53±10.94 vs. 71.07±4.51, t= 12.802, P= 0.000). Conclusion: Lower expression of miR-124 is seen in gastric cancer tissues,suggesting that this gene may inhibit the proliferation and migration yet promote apoptosis of gastric cancer cells. The regulatory mechanisms may be involved in targeted PI3K/AKT signaling pathway.
作者
符清胜
吴克盛
赵军
赵国海
FU Qingsheng;WU Kesheng;ZHAO Jun;ZHAO Guohai(Department of Gastrointestinal Surgery,The First Affiliated Hospital of Wannan Medical College,Wuhu 241001,China)
出处
《皖南医学院学报》
CAS
2019年第5期414-418,共5页
Journal of Wannan Medical College
基金
安徽重点研究和开发计划项目(1804h08020242)
