miR-373靶向SIRT1基因抑制胃癌细胞增殖和迁移的研究  

miR-373 inhibits proliferation and migration of gastric cancer cells by targeting SIRT1

作　　者：刘闯 周昕[2] 王飞[2] LIU Chuang;ZHOU Xin;WANG Fei(Department of Internal Medicine,Dahua Hospital,Xuhui District,Shanghai 200237,China;College of Medicine,Anhui University of Science and Technology,Huainan 232000,China)

机构地区：[1]上海市徐汇区大华医院内科,上海200237 [2]安徽理工大学医学院,淮南232000

出　　处：《中国肿瘤临床与康复》2022年第10期1174-1179,共6页Chinese Journal of Clinical Oncology and Rehabilitation

摘　　要：目的 探讨miR-373抑制胃癌细胞增殖、迁移的研究作用机制。方法 将miR-373模拟物转至胃癌细胞后,采用MTT实验检测细胞增殖能力。采用荧光素酶报告实验miR-373与SIRT1基因的靶向关系。采用RT-PCR实验检测细胞中SIRT1基因表达水平,Transwell实验检测细胞迁移能力。采用流式细胞术检测miR-373对细胞的周期影响。采用Western blotting实验检测胃癌细胞基质金属蛋白酶9(MMP-9)、c-Myc及Cyclin D1表达水平。结果 miR-373组的miR-373的相对表达水平为(3.16±0.27),高于空白对照及阴性对照,差异有统计学意义(P<0.05)。荧光素酶报告基因实验发现,转染miR-373后可降低SIRT1野生质粒的荧光素酶活性水平,差异有统计学意义(P<0.05)。与空白对照组及阴性对照相比,miR-373组的SIRT1的mRNA表达水平下降,差异有统计学意义(P<0.05)。与空白对照组及阴性对照相比,miR-373组的胃癌细胞的增殖水平下降,差异有统计学意义(P<0.05)。流式细胞术实验发现,miR-373组的G/G期比例为(57.36±4.52)%,低于空白对照组及阴性对照,S期比例为(31.72±2.68)%,高于空白对照组及阴性对照,差异有统计学意义(P<0.05)。Transwell实验发现,miR-373组的迁移细胞数量(42.36±5.27)个,低于空白对照组及阴性对照,差异有统计学意义(P<0.05)。Western blot实验发现,miR-373组MMP-9、c-Myc及Cyclin D1蛋白相对水平为(0.16±0.03)、(0.18±0.04)、(0.12±0.03),低于空白对照组及阴性对照组,差异有统计学意义(P<0.05)。结论 miR-373可靶向抑制SIRT1基因表达,从而干扰胃癌细胞的增殖、迁移能力。Objective To investigate the mechanism of the suppressive effect of miR-373 on the proliferation and migration of gastric cancer cells.MethodsAfter the miR-373 mimic was transferred to gastric cancer line MGC-803,MTT assay was used to detect the proliferation level of MGC-803 cells.Luciferase reporter assay was used to detect the targeting relationship between miR-373 and SIRT1 gene.Transwell assay was used to detect the cell migration ability.Flow cytometry was used to detect the effect of miR-373 on the cell cycle of MGC-803 cells.Western blotting assay was used to detect the expression of matrix metallopeptidase 9(MMP-9),c-Myc and Cyclin D1 in gastric cancer cells.ResultsThe relative expression level of miR-373 in the group of miR-373 was 3.16±0.27,which was significantly higher than that in the blank control group and negative control group(P<0.05).Luciferase reporter gene experiment showed that the luciferase activity of SIRT1 wild plasmid was significantly decreased after transfection of miR-373(P<0.05).Compared with the blank control group and the negative control group,the expression of SIRT1 in the miR-373 group decreased significantly(P<0.05).Compared with the blank control group and the negative control group,the proliferation level of gastric cancer cells in the miR-373 group decreased significantly(P<0.05).Flow cytometry showed that the ratio of G_(0)/G_(1)phase was 57.36%±4.52%in the miR-373 group,which was significantly lower than that in the blank control group and the negative control group,and the ratio of S phase was 31.72%±2.68%which was significantly higher than that in the blank control group and the negative control group(all P<0.05).Transwell experiment showed that the number of migrating cells was 42.36±5.27 in the group of miR-373,which was significantly lower than that in the blank control group and the negative control group(all P<O.05).Western blot assay showed that the relative levels of MMP-9,c-Myc and Cyclin D1 proteins in the miR-373 group were 0.16±0.03,0.18±0.04 and 0.12±

关 键 词：miR-373 SIRT1基因 增殖 迁移 

分 类 号：R735.2[医药卫生—肿瘤]