Mir-335-5p靶向G6PD对结肠癌细胞增殖、凋亡的影响  

Effects of miR-335-5p targeting G6PD on proliferation and apoptosis of colon cancer cells

作　　者：樊小群 李欢[1] 朱华雄 黄健平 何吕芬 FAN Xiao-qun;LI Huan;ZHU Hua-xiong;HUANG Jian-ping;HE Lyu-fen(Department ofLaboratory,Sanya People's Hospital,Sanya 572000,China)

机构地区：[1]海南省三亚市人民医院检验科,三亚572000

出　　处：《中国应用生理学杂志》2021年第4期402-406,共5页Chinese Journal of Applied Physiology

摘　　要：目的:探讨Mir-335-5p通过靶向G6PD对结肠癌细胞增殖、凋亡的影响。方法:设置正常结肠细胞组、空白对照组、NC组、miRNA-335-5p mimic组;体外培养结肠上皮细胞(IEC)和人源性结肠癌细胞SW480,并对NC组、miRNA-335-5p mimic组细胞进行转染;采用RT-qPCR检测各组细胞中miR-335-5p及G6PD mRNA表达水平;双荧光素酶报告实验验证Mir-335-5p对G6PD靶向作用;MTT实验检测各组细胞的增殖;流式细胞术检测转染后各组细胞凋亡率;Western blot检测G6PD及凋亡蛋白Bax、Bcl-2、caspase3相对表达水平。结果:与正常结肠细胞相比,空白对照组、NC组结肠癌细胞SW480细胞中miR-335-5p相对表达水平降低,G6PD mRNA相对表达水平升高(P<0.05);与空白对照组、NC组相比,miR-335-5p mimic组细胞miR-335-5p表达水平明显升高,G6PD mRNA表达水平显著降低(P<0.05)。与空白对照、NC组相比,miR-335-5p mimic组结肠癌SW480细胞生长活性显著降低,凋亡率显著升高(P<0.05)。miR-335-5p mimic+WT-G6PD 3′-UTR组的荧光素酶相对活性低于miR-335-5p NC+WT-G6PD 3′-UTR组(P<0.05)。与空白对照组相比,miR-335-5p mimic组细胞中G6PD、Bcl-2蛋白相对表达水平显著降低,Bax、caspase3蛋白表达水平显著升高(P<0.05)。结论:Mir-335-5p可能通过靶向G6PD抑制结肠癌细胞增殖、促进凋亡。Objective:To investigate the effects of miR-335-5p targeting glucose-6-phosphate dehydrogenase(G6PD)on the proliferation and apoptosis of colon cancer cells.Methods:Normal colon cell group,blank control group,NC group and miRNA-335-5p mimic group were set up.Colonic epithelial cells(IEC)and human colon cancer cells SW480 were cultured in vitro,and the cells in the NC group and miRNA-335-5p mimic group cells were transfected.RT-qPCR was used to detect the expression levels of miR-335-5p and G6PD mRNA in each group of cells.The targeting effect of miR-335-5p on G6PD was verified by Double Luciferase Report experiment.MTT assay was used to detect cell proliferation.Flow cytometry was used to detect the apoptosis rate.The expressions of G6PD,Bax,Bcl-2 and caspase-3 were detected by Western blot.Results:Compared with normal colon cells,the relative expression levels of miR-335-5p in SW480 cells of colon cancer in the blank control group and NC group were decreased,and the relative expression level of G6PD mRNA was increased(P<0.05);compared with the blank control group and NC group,the expression level of miR-335-5p in miR-335-5p mimic group was increased significantly,and the expression of G6PD mRNA was decreased significantly(P<0.05).Compared with the blank control group and NC group,the proliferative activity of colon cancer SW480 cells in miR-335-5p mimic group was decreased significantly,and the apoptosis rate was increased significantly(P<0.05).The relative activity of luciferase in miR-335-5p mimic+WT-G6PD 3'-UTR group was lower than that in miR-335-5p NC+WT-G6PD 3'-UTR group(P<0.05).Compared with the blank control group,the relative expression levels of G6PD and bcl-2 protein in miR-335-5p mimic group were decreased significantly,and the expression levels of Bax and caspase-3 protein were increased significantly(P<0.05).Conclusion:MiR-335-5p may inhibit the proliferation and promote apoptosis of colon cancer cells by targeting G6PD.

关 键 词：microRNA-335-5p 6-磷酸葡萄糖脱氢酶 结肠癌 细胞增殖 细胞凋亡 

分 类 号：R363[医药卫生—病理学] R735.3[医药卫生—基础医学]