miRNA-186上调Dicer1的表达影响胃癌细胞侵袭和迁移的作用机制研究  

Mechanism of miRNA-186 up-regulation of the expression of Dicerl and affecting the invasion and migration of gastric cancer cell

作　　者：李道宏 胡爱侠[1] 靳钰炜 LI Daohong;HU Aixia;JIN Yuwei(Department of Pathology,He’nan Provincial People’s Hospital,Zhengzhou 450000,He’nan,China)

机构地区：[1]河南省人民医院病理科,郑州4500000

出　　处：《癌症进展》2021年第9期891-895,共5页Oncology Progress

摘　　要：目的探讨miRNA-186上调Dicer1的表达影响胃癌细胞侵袭和迁移的作用机制。方法取40例胃癌患者的胃癌组织及相应的癌旁组织;将转染pLVTHM-mimic-miRNA-186和pLVTHM-mimic-NC的慢病毒感染液分别转染BGC-823细胞,得到过表达BGC-823-mimics-miRNA-186细胞、阴性对照BGC-823-mimics-NC细胞。定量逆转录聚合酶链反应(qRT-PCR)检测不同组织和细胞中miRNA-186的相对表达量;Transwell小室检测细胞侵袭能力,划痕实验检测细胞迁移能力;间接免疫荧光法(IFA)检测Dicer1相关分子神经型钙黏蛋白(N-cadherin)的阳性表达情况,双荧光素酶报告基因实验验证miRNA-186和Dicer1的靶向关系,Western blot检测各细胞Dicer1的相对表达量。结果胃癌组织中miRNA-186的相对表达量明显低于癌旁组织(P﹤0.01)。胃癌细胞BGC-823和BGC-823-mimics-NC中miRNA-186的相对表达量均明显低于正常胃黏膜上皮细胞RGM-1,且BGC-823-mimics-miRNA-186细胞中miRNA-186的相对表达量明显高于胃癌细胞BGC-823及BGC-823-mimics-NC,差异均有统计学意义(P﹤0.01)。BGC-823-mimics-miRNA-186细胞侵袭数目少于胃癌细胞BGC-823和BGC-823-mimics-NC,细胞迁移率低于胃癌细胞BGC-823和BGC-823-mimics-NC,差异均有统计学意义(P﹤0.05)。过表达miRNA-186的BGC-823-mimics-miRNA-186细胞中N-cadherin阳性表达率低于胃癌细胞BGC-823和BGC-823-mimics-NC(P﹤0.05)。mimics-miRNA-186能明显抑制野生型psi-CHECK-Dicer1-wild的荧光素酶活性,而对突变型psi-CHECK-Dicer1-mutant的荧光素酶活性无明显影响。胃癌细胞BGC-823和BGC-823-mimics-NC中Dicer1蛋白的相对表达量均明显低于正常胃黏膜上皮细胞RGM-1,且BGC-823-mimics-miRNA-186细胞中Dicer1的相对表达量明显高于胃癌细胞BGC-823和BGC-823-mimics-NC,差异均有统计学意义(P﹤0.01)。结论miRNA-186通过靶向正调控Dicer1的表达,下调N-cadherin的表达可抑制胃癌细胞的侵袭和迁移能力。Objective To investigate the effect of miRNA-186 on invasion and metastasis of gastric cancer cells through Dicer1 up-regulation and its underlying mechanism.Method A total of 40 patients with gastric cancer were selected,and their gastric cancer tissues and paracancerous tissues were collected.The lentiviral infection solution transfected with pLVTHM-mimic-miRNA-186 and pLVTHM-mimic-NC were respectively transfected into BGC-823 cells to obtain over-expressing BGC-823-mimics-miRNA-186 cells and negative control BGC-823-mimics-NC cells.The relative expression of miRNA-186 in different tissues and cells was detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Transwell chamber detects cell invasion ability,cell migration ability was measured by cell scratch test,the positive rate of Dicer1 related molecule N-cadherin was detected by indirect immunofluorescence assay(IFA),the targeting relationship between miRNA-186 and Dicer1 was verified by the dual-luciferase reporter gene experiment,and the relative expression of Dicer1 in each cell was detected by Western blot.Result The relative expression of miRNA-186 in gastric cancer tissue was significantly lower than that in adjacent tissues(P<0.01).The relative expression of miRNA-186 in gastric cancer cells BGC-823 and BGC-823-mimics-NC was significantly lower than that of normal gastric epithelial cells RGM-1,and the relative expression of miRNA-186 in BGC-823-mimics-miRNA-186 cells was significantly higher than that of gastric cancer cells BGC-823 and BGC-823-mimics-NC,and the differences were statistically significant(P<0.01).The number of invasions of BGC-823-mimics-miRNA-186 cells was less than that of gastric cancer cells BGC-823 and BGC-823-mimics-NC,and the cell migration rate was lower than that of gastric cancer cells BGC-823 and BGC-823-mimics-NC,the differences were statistical significance(P<0.05).The positive rate of N-cadherin in BGC-823-mimics-miRNA-186 cells with miRNA-186 overexpressing was lower than that in gastric cancer

关 键 词：miRNA-186 胃癌 Dicer1 侵袭 迁移 

分 类 号：R735.2[医药卫生—肿瘤]