miR-641表达对肺腺癌A549细胞增殖和侵袭及迁移能力影响  

Effect of miR-641 expression on the proliferation,invasion and migration of lung adenocarcinoma A549 cells

作　　者：杨扬[1] 王凯 任清泉[1] 许瑞彬[1] 郭建峰[1] YANG Yang;WANG Kai;REN Qing-quan;XU Rui-bin;GUO Jian-feng(Department of Thoracic Surgery,Affiliated Hospital of Yan’an University,Yan'an 716000,China)

机构地区：[1]延安大学附属医院胸外科,陕西延安716000

出　　处：《中华肿瘤防治杂志》2021年第21期1611-1617,共7页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的探讨miR-641在肺腺癌中的表达情况及其分子生物学作用机制。方法收集延安大学附属医院胸外科2018-01-01-2019-03-31行手术切除的30例肺腺癌患者癌组织及对应周围正常肺组织标本,采用qPCR法检测肺腺癌组织和肺腺癌细胞系A549、H1299和H157中miR-641表达水平,分析miR-641表达与肺腺癌患者临床特征关系。MTT法检测miR-641对肺腺癌A549细胞增殖能力的影响;Transwell侵袭及迁移实验检测miR-641对肺腺癌A549细胞迁移及侵袭能力的影响;靶基因双荧光素酶实验检测miR-641与CYR61的相互作用;PCR和蛋白质印迹法检测A549细胞中CYR61表达。结果 qPCR法检测miR-641在肺腺癌组织、肺组织、A549、H1299、H157和WI-38细胞中表达值分别为0.14±0.02、1.01±0.08、0.35±0.04、0.47±0.05、0.48±0.04、0.98±0.08,F=775.537,P<0.001;肺腺癌组织和肺组织miR-641表达差异有统计学意义,t=57.786,P<0.001。miR-641表达与TNM分期(P=0.004)和淋巴结转移(P=0.013)有关联。NC组和miR-641 mimics组A549细胞增殖活力分别为0.98±0.07和8.23±0.53,t=23.489,P<0.001。miR-641mimics转染A549细胞24、48和72h后,细胞增殖活力分别为1.23±0.19、1.86±0.21和3.05±0.29,F=333.197,P<0.001。Transwell侵袭及迁移实验结果显示,与NC组(101.42±10.14)%和(97.24±7.63)%相比,miR-641mimics组迁移及侵袭力分别为(71.43±6.12)%和(63.47±7.12)%,t值分别为13.620和12.430,均P<0.001。Target Sacn生物信息网站预测miR-641基因序列与CYR61基因序列存在结合片段,靶基因双荧光素酶实验显示,野生型CYR61 3′-UTR相对双荧光素酶活性低于突变型CYR61 3′-UTR,F=49.832,P<0.001。NC组和miR-641mimics组CYR61蛋白表达水平分别为1.00±0.09和0.61±0.10,t=5.024,P=0.004。MTT增殖实验显示,与miR-641mimics相比较,miR-641mimics+pSUPER-CYR61转染A549细胞24、48和72h后细胞增殖活性增加,分别为1.61±0.15、2.29±0.23和3.46±0.34,F=356.301,P<0.001。Transwell迁移实验表明,NC组、miR-641mimicsObjective To explore the expression of miR-641 in lung adenocarcinoma and its molecular biological mechanism.Methods A total of 30 cases of resected lung adenocarcinoma and corresponding normal lung tissues between January 1,2018 and March 31,2019 were involved in this study.The expressions of miR-641 in lung adenocarcinoma tissues,normal lung tissues as well as A549,H1299 and H157 cell lines were detected by qPCR.The relationship between the expression of miR-641 and the clinical features of lung adenocarcinoma was investigated.MTT assay was used to detect the effect of miR-641 on the proliferation of A549 cells.Transwell invasion and migration assay was used to detect the effect of miR-641 on the migration and invasion of A549 cells.Dual luciferase reporter assay was used to detect the interaction between miR-641 and CYR61.PCR and western blot were used to detect the expression of CYR61 in A549 cells.Results The expressions of miR-641 in lung adenocarcinoma,lung tissue,A549,H1299,h157 and WI-38 cells were0.14±0.02,1.01±0.08,0.35±0.04,0.47±0.05,0.48±0.04 and 0.98±0.08,respectively(F=775.537,P<0.001).There was significant difference in the mean expression of miR-641 between lung adenocarcinoma and lung tissue(t=57.786,P<0.001).The miR-641 expression was correlated with TNM stage(P=0.004)and lymph node metastasis(P=0.013).The proliferation activity of A549 cells in NC group and miR-641 mimics group were 0.98 ±0.07 and8.23±0.53(t=23.489,P<0.001).After transfection of A549 cells with miR-641 mimics for 24,48 and 72 h,the cell proliferation activities were 1.23±0.19,1.86±0.21 and 3.05±0.29,respectively(F=333.197,P<0.001).The results of Transwell invasion and migration experiment showed that compared with NC group(101.42±10.14)% and(97.24±7.63)%,the migration and invasion of miR-641 mimics group were(71.43±6.12)% and(63.47±7.12)%,respectively(t values were 13.620 and 12.430,all P<0.001).Target sacn bioinformatics website predicted the binding fragment between miR-641 gene sequence and CYR61 gene sequenc

关 键 词：肺腺癌 miR-641 CYR61 增殖 侵袭 迁移 

分 类 号：R734.2[医药卫生—肿瘤]