miR-144通过靶向TIGAR调控胃癌细胞增殖与凋亡及自噬的分子机制  

Molecular mechanisms of miR-144 regulating proliferation, apoptosis and autophagy of gastric cancer cells by targeting TIGAR

作　　者：谭业儒[1] 伍小平[1] 李跃华[1] TAN Ye-ru;WU Xiao-ping;LI Yue-hua(Department of Internal Medicine-Oncology,The First Affiliated Hospital of University of South China,Hengyang 421001,Hunan Province,China)

机构地区：[1]南华大学附属第一医院肿瘤内科

出　　处：《中国临床药理学杂志》2020年第1期57-60,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究miR-144对胃癌细胞增殖、凋亡和自噬的影响和机制。方法将miR-144 mimics、mimics control分别转染胃癌细胞,依次标记为mimics-NC组、miR-144 mimics组。将miR-144 mimics分别与pcDNA3.1、pcDNA3.1-TIGAR共转染至胃癌细胞,依次标记为miR-144 mimics+NC组、miR-144 mimics+TIGAR组。正常培养的胃癌细胞标记为Control组。以噻唑蓝法检测细胞增殖,以流式细胞术测定细胞凋亡,以Western blot检测凋亡蛋白caspase-3和自噬蛋白Beclin1、LC3Ⅱ/Ⅰ表达变化,以生物信息学软件预测miR-144的靶基因可能为TIGAR,利用荧光素酶报告系统鉴定靶向关系。结果 mimics-NC组和miR-144 mimics组的细胞增殖能力分别为0.36±0.05,0.20±0.02,细胞凋亡率分别为3.05±0.34,13.84±1.47,caspase-3表达水平分别为0.22±0.04,0.43±0.05, Beclin1分别为0.38±0.06,0.73±0.07,LC3Ⅱ分别为0.19±0.03,0.39±0.05,差异均有统计学意义(均P<0.05)。miR-144 mimics+NC组和miR-144 mimics+TIGAR组的细胞增殖能力分别为0.21±0.04,0.32±0.03,细胞凋亡率分别为12.04±1.25,5.98±0.73,caspase-3表达水平分别为0.40±0.03,0.24±0.05,Beclin1分别为0.60±0.05,0.46±0.06,LC3Ⅱ分别为0.34±0.06,0.20±0.03,差异均有统计学意义(均P<0.05)。结论 miR-144靶向抑制TIGAR表达降低胃癌细胞增殖能力,并诱导胃癌细胞凋亡和自噬。Objective To study the effect and mechanism of miR-144 on proliferation, apoptosis and autophagy of gastric cancer cells. Methods miR-144 mimics and mimics control were transfected into gastric cancer cells, which were labeled as mimics-NC group and miR-144 mimics group. miR-144 mimics were co-transfected with pcDNA3.1 and pcDNA3.1-TIGAR into gastric cancer cells, which were labeled as miR-144 mimics+NC group and miR-144 mimics+TIGAR group. Normally cultured gastric cancer cells were labeled as Control group. MTT was used to detect cell proliferation, cell apoptosis was measured by flow cytometry, Western blot was used to detect the expression of apoptotic protein caspase-3 and autophagic protein Beclin1 and LC3 Ⅱ/Ⅰ. Bioinformatics software predicts that the target gene of miR-144 may be TIGAR, luciferase reporting system was used to identify the targeting relationship. Results The cell proliferation in mimics-NC group and miR-144 mimics group were 0. 36 ± 0. 05,0. 20 ± 0. 02,the apoptotic rates were 3. 05 ± 0. 34,13. 84 ± 1. 47,the expression of apoptotic protein caspase-3 were 0. 22 ± 0. 04,0. 43 ± 0. 05,the levels of Beclin1 were 0. 38 ± 0. 06,0. 73 ± 0. 07,LC3Ⅱwere 0. 19 ± 0. 03,0. 39 ± 0. 05,all with significant difference( all P < 0. 05).The cell proliferation in miR-144 mimics + NC group and miR-144 mimics + TIGAR group were 0. 21 ± 0. 04,0. 32 ± 0. 03,the apoptotic rates were 12. 04 ± 1. 25,5. 98 ± 0. 73,the caspase-3 were 0. 40 ± 0. 03,0. 24 ± 0. 05,the protein levels of Beclin1 were 0. 60 ± 0. 05,0. 46 ± 0. 06,LC3Ⅱ were 0. 34 ± 0. 06,0. 20 ± 0. 03,all with significant difference( all P < 0. 05). Conclusion miR-144 targeting inhibited TIGAR expression,reduced proliferation of gastric cancer cells,and induces apoptosis and autophagy of gastric cancer cells.

关 键 词：胃癌细胞 自噬 凋亡 TP53诱导的糖酵解和凋亡调节因子 

分 类 号：R979.1[医药卫生—药品]