二甲双胍联合miR-145对胃癌BGC823细胞增殖凋亡及AKT信号通路的影响  

Effects of metformin combined with miR-145 on proliferation,apoptosis and AKT signaling pathway in gastric cancer BGC823 cells

作　　者：田春阳[1] 刘晓政[1] 范军朝 TIAN Chunyang;LIU Xiaozheng;FAN Junchao(Department of Gastroenterology,Nanyang Central Hospital,Nanyang,Henan,China,473000;Department of Anesthesiology,the First Affiliated Hospital of Henan University,Zhengzhou,Henan,China,475000)

机构地区：[1]南阳市中心医院消化内科,河南南阳473000 [2]河南大学第一附属医院麻醉科,河南郑州475000

出　　处：《分子诊断与治疗杂志》2021年第9期1498-1502,共5页Journal of Molecular Diagnostics and Therapy

基　　金：河南省教育厅科研基金项目(16B320003)。

摘　　要：目的探讨二甲双胍联合miR-145对胃癌BGC823细胞增殖凋亡的影响及其机制。方法将miR-145 mimic及其阴性对照转染至胃癌BGC823细胞中并记为miR-145组和NC组,并以未转染细胞作为对照组,RT-PCR检测细胞中miR-145的表达;MTT法检测不同浓度(0、2.5、5、10、20和40mmol/l)二甲双胍对BGC823细胞增殖的影响,并并筛选出合适的作用浓度。将BGC823细胞随机分为未处理组(未做处理)、二甲双胍组(给予10 mmol/L二甲双胍处理72 h)、miR-145组(转染miR-145 mimics)和二甲双胍+miR-145组(转染miR-145 mimics后,给予10 mmol/L二甲双胍处理72 h),采用MTT法和流式细胞仪分别检测各组细胞中存活率和凋亡率,Western blot检测细胞中CyclinD1、Bcl-2和p-AKT蛋白的表达。结果与对照组相比,miR-145组细胞中miR-145的表达明显升高(P<0.05),而NC组细胞中miR-145表达与对照组比较差异无统计学意义(P>0.05);与空白对照组(0 mmol/L)相比,二甲双胍处理组中细胞的存活率均显著降低(P<0.05),且呈时间-剂量依赖性。与未处理组相比,二甲双胍组、miR-145组和二甲双胍+miR-145组细胞存活率明显降低,凋亡率明显升高(P<0.05),细胞中CyclinD1、Bcl-2和p-AKT蛋白显著降低,且二甲双胍+miR-145组的作用强度明显大于二甲双胍组或miR-145组(P<0.05)。结论上调miR-145表达可增强二甲双胍抑制胃癌BGC823细胞增殖促进细胞凋亡的作用,其分子机制可能与抑制AKT信号通路有关。Objective To investigate the effect of metformin combined with miR-145 on proliferation and apoptosis of gastric cancer BGC823 cells and its mechanism.Methods MiR-145 mimic and its negative control were transfected into BGC823 cells of gastric cancer and recorded as miR-145 group and NC group,and untransfected cells as control group,the expression of miR-145 in cells was detected by RT-PCR.The effect of different concentrations(0,2.5,5,10,20 and 40 mmol/L) of metformin on the proliferation of BGC823 cells was tested by MTT method and the appropriate concentration was selected.BGC823 cells were randomly divided into untreated group(untreated),metformin group(treated by 10 mmol/L metformin for 72 h),miR-145 mimics group(transfected with miR-145 mimics) and metformin+miR-145 group(after transfection of miR-145 mimics,treated by 10 mmol/L metformin for 72 h),the survival rate and apoptosis rate in each group were separatly measured by MTT and flow cytometry,and the expressions of CyclinDl,Bcl-2 and p-AKT proteins in the cells were checked by Western blot.Results Compared with the control group,the expression of miR-145 was significantly higher in the miR-145 group(P<0.05),but there was no significant difference between the NC group and the control group(P>0.05);compared with the blank control group(0 mmol/L),the survival rate of cells in the metformin group was significantly lower(P<0.05),and in time-dose-dependent.Compared with the untreated group,the cell survival rate of metformin group,miR-145 group and metformin+miR-145 group were decreased significantly,the apoptosis was increased significantly,and the expressions of CyclinDl,Bcl-2 and p-AKT proteins were reduced significantly,and the action intensity of metformin+miR-145 group was significantly greater than that of metformin group or miR-145 group(P<0.05).Conclusion Up regulation of miR-145 can enhance the effect of metformin on the proliferation and apoptosis of gastric cancer BGC823 cells,and its molecular mechanism may be related to inhibition of AKT signal

关 键 词：胃癌 二甲双胍 MIR-145 细胞增殖 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]