Fan Xiao Qun, Li Huan, Zhu Hua Xiong, Huang Jian Ping, He Lyu Fen
Department ofLaboratory, Sanya People's Hospital, Sanya 572000, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2021 Jul;37(4):402-406. doi: 10.12047/j.cjap.6090.2021.041.
To investigate the effects of miR-335-5p targeting glucose-6-phosphate dehydrogenase (G6PD) on the proliferation and apoptosis of colon cancer cells. 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. 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 (＜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 (＜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 (＜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 (＜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 (＜0.05). MiR-335-5p may inhibit the proliferation and promote apoptosis of colon cancer cells by targeting G6PD.
探讨miR-335-5p靶向葡萄糖-6-磷酸脱氢酶（G6PD）对结肠癌细胞增殖和凋亡的影响。设立正常结肠细胞组、空白对照组、阴性对照组（NC组）和miRNA-335-5p模拟物组。体外培养结肠上皮细胞（IEC）和人结肠癌细胞SW480，并对NC组和miRNA-335-5p模拟物组细胞进行转染。采用RT-qPCR检测各组细胞中miR-335-5p和G6PD mRNA的表达水平。通过双荧光素酶报告实验验证miR-335-5p对G6PD的靶向作用。采用MTT法检测细胞增殖情况。采用流式细胞术检测凋亡率。通过蛋白质免疫印迹法检测G6PD、Bax、Bcl-2和caspase-3的表达。与正常结肠细胞相比，空白对照组和NC组结肠癌SW480细胞中miR-335-5p的相对表达水平降低，G6PD mRNA的相对表达水平升高（P＜0.05）；与空白对照组和NC组相比，miRNA-335-5p模拟物组中miR-335-5p的表达水平显著升高，G6PD mRNA的表达显著降低（P＜0.05）。与空白对照组和NC组相比，miRNA-335-5p模拟物组中结肠癌SW480细胞的增殖活性显著降低，凋亡率显著升高（P＜0.05）。miR-335-5p模拟物+野生型G6PD 3'-UTR组中荧光素酶的相对活性低于miR-335-5p阴性对照+野生型G6PD 3'-UTR组（P＜0.05）。与空白对照组相比，miRNA-335-5p模拟物组中G6PD和bcl-2蛋白的相对表达水平显著降低，Bax和caspase-3蛋白的表达水平显著升高（P＜0.05）。miR-335-5p可能通过靶向G6PD抑制结肠癌细胞的增殖并促进其凋亡。
