摘要
目的:探究miRNA-216b(miR-216b)对顺铂处理的胃癌细胞的影响及其可能的作用机制。方法:体外培养人胃黏膜细胞系GES-1、人胃癌细胞系SGC7901和胃癌顺铂耐药细胞系SGC7901/DDP,将SGC7901/DDP细胞随机分为空白对照组、agomir-NC+oe-NC组、agomir-miR-216b+oe-NC组、agomir-miR-216b+oe-Beclin1组,按照分组进行细胞转染,荧光素酶报告实验检测miR-216b与Beclin1的靶向结合;qPCR检测细胞miR-216b的表达;Western blot检测细胞Beclin1、LC3、p62、Bax和Bcl-2蛋白表达;CCK-8检测细胞活力;克隆形成实验检测细胞增殖;流式细胞术检测细胞凋亡。结果:与GES-1组比较,SGC7901组细胞中miR-216b的表达显著降低(P<0.05),Beclin1蛋白表达显著升高(P<0.05);与SGC7901组比较,SGC7901/DDP组细胞中miR-216b的表达显著降低(P<0.05),Beclin1蛋白表达显著升高(P<0.05)。miR-216b靶向调控Beclin-1,与空白对照组比较,agomir-NC+oe-NC组SGC7901/DDP细胞中各指标差异无统计学意义;与agomir-NC+oe-NC组比较,agomir-miR-216b+oe-NC组SGC7901/DDP细胞中miR-216b的表达显著升高(P<0.05),Beclin1、LC3Ⅱ/LC3Ⅰ和Bcl-2蛋白表达显著降低(P<0.05),细胞活力和克隆数显著降低(P<0.05),细胞凋亡率、Bax和p62蛋白表达显著升高(P<0.05);与agomir-miR-216b+oe-NC组比较,agomir-miR-216b+oe-Beclin1组SGC7901/DDP细胞中Beclin1、LC3Ⅱ/LC3Ⅰ和Bcl-2蛋白表达显著升高(P<0.05),细胞活力和克隆数显著升高(P<0.05),细胞凋亡率、Bax和p62蛋白表达显著降低(P<0.05)。结论:miR-216b通过靶向调控Beclin1抑制自噬并促进凋亡,增加SGC7901/DDP细胞顺铂敏感性。
Objective: To explore the effect of miR-216 b on gastric cancer cells treated with cisplatin and its possible mechanism. Methods: Human gastric mucosal cell line GES-1, human gastric cancer cell line SGC7901 and gastric cancer cisplatin-resistant cell line SGC7901/DDP were cultured in vitro. SGC7901/DDP cells were randomly divided into blank control group, agomir-NC+oe-NC group, agomir-miR-216 b+oe-NC group, agomir-miR-216 b+oe-Beclin1 group, the cells were transfected according to the groups, and the luciferase reporter experiment was used to detect the targeted binding of miR-216 b and Beclin1;qPCR was used to detect the expression of miR-216 b in cells;Western blot was used to detect the expression of Beclin1, LC3, p62, Bax, and Bcl-2 proteins in cells;CCK-8 was used to detect cell viability;clone formation experiments were used to detect cell proliferation;flow cytometry was used to detect apoptosis. Results: Compared with the GES-1 group, the expression of miR-216 b in the cells of the SGC7901 group was significantly reduced(P<0.05), and the expression of Beclin1 protein was significantly increased(P<0.05);compared with the SGC7901 group, the expression of miR-216 b in the cells of the SGC7901/DDP group was significantly reduced(P<0.05), and the expression of Beclin1 protein was significantly increased(P<0.05). miR-216 b regulates Beclin-1 expression through targeted binding. Compared with the blank control group, there was no statistically significant difference in the indicators of SGC7901/DDP cells in the agomir-NC+oe-NC group(P>0.05);compared with agomir-NC+oe-NC group, the expression of miR-216 b in SGC7901/DDP cells in the agomir-miR-216 b+oe-NC group was significantly increased(P<0.05), and the expression of Beclin1, LC3Ⅱ/LC3Ⅰ and Bcl-2 protein was significantly decreased(P<0.05), cell viability and clone number were significantly reduced(P<0.05), cell apoptosis rate, expression of Bax and p62 protein were significantly increased(P<0.05);compared with agomir-miR-216 b+oe-NC group, the expression of Beclin1, LC3Ⅱ/LC3Ⅰ and Bcl-2 protein in SGC7901/DDP cells in agomir-miR-216 b+oe-Beclin1 group was significantly increased(P<0.05), cell viability and clone number were significantly increased(P<0.05), cell apoptosis rate, expression of Bax and p62 protein were significantly reduced(P<0.05). Conclusion: miR-216 b inhibits autophagy and promotes apoptosis through targeted regulation of Beclin1 to increase the cisplatin sensitivity of SGC7901/DDP cells.
作者
周京涛
刘佳
李亮
李建刚
努尔买买提·阿米都拉
闫军
寇旭东
艾孜买提·艾海提
马博
ZHOU Jingtao;LIU Jia;LI Liang;LI Jiungang;Nurmumat Atnidulu;YAN Ju;KOU Xudong;Aizmat Aiheti;MA Bo(Department of General Surgery,Seventh Affiliated Hospital of Xinjiang Medical University,Urumqi,830000,China;Department of Emergency,Eighth People's Hospital of Xinjiang Uygur Autonomous Region;Department of General Surgery,Second Affiliated Hospital of Xinjiang Medical University)
出处
《中国中西医结合消化杂志》
CAS
2021年第6期398-405,共8页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
基金
新疆维吾尔自治区卫生健康青年医学科技人才专项科研项目(No:WJWY-202006)。
作者简介
通信作者:马博,E-mail:maboyang@sina.com。
