miR-671-5p对非小细胞肺癌细胞增殖和迁移及侵袭影响及其作用机制  

Effects and mechanism of miR-671-5p on proliferation,migration and invasion of non-small cell lung cancer cells

作　　者：许英杰 张艳炜 李海洲 张磊[1] 杨门 韩京军[1] XU Ying-jie;ZHANG Yan-wei;LI Hai-zhou;ZHANG Lei;YANG Men;HAN Jing-jun(Department of Thoracic and Cardiovascular Surgery,Eighth Affiliated Hospital,Sun Yat-sen University,Shenzhen 518033,China;Department of Immune-PLanning,Shenzhen Center for Disease Control and PreventionnShenzhen 518055,China)

机构地区：[1]中山大学附属第八医院胸心外科,广东深圳518033 [2]深圳市疾病预防和控制中心免疫规划科,广东深圳518055

出　　处：《中华肿瘤防治杂志》2021年第20期1541-1548,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81402755);广东省医学科学技术研究基金(A2020168);深圳市卫生计生科研项目(SZGW2017003)。

摘　　要：目的探讨miR-671-5p调控非小细胞肺癌细胞系增殖、迁移和侵袭的作用机制。方法采用实时荧光定量PCR(RT-qPCR)法检测人正常肺上皮细胞系(BEAS-2B)和非小细胞肺癌细胞系(A549和H460)中miR-671-5p及其靶基因Kruppel样锌指转录因子6(KLF6)mRNA表达量。miR-671-5p mimic转染A549和H460细胞,设置阴性对照序列,蛋白质印迹法检测KLF6蛋白表达,细胞计数试剂盒(CCK-8)法、EdU法和Transwell小室法分别检测非小细胞肺癌细胞的增殖、迁移和侵袭能力。结果与BEAS-2B细胞比较,miR-671-5p在A549和H460细胞中高表达,表达量分别为1.646±0.026和1.638±0.027,差异有统计学意义,F=153.900,P<0.001;KLF6在A549和H460中的mRNA表达量降低,分别为0.233±0.005和0.382±0.003,差异有统计学意义,F=2 077.000,P<0.001;转染miR-671-5p mimic后,A549和H460细胞的增殖、迁移和侵袭能力增强(均P<0.05),KLF6的mRNA和蛋白表达水平受到抑制,其中A549和H460细胞KLF6蛋白水平分别下调至0.252±0.011(t=23.970,P<0.001)和0.290±0.017(t=16.140,P<0.001)。结论 miR-671-5p可能通过调控KLF6基因促进非小细胞肺癌细胞系的增殖、迁移及侵袭,进而影响非小细胞肺癌的发生和发展。Objective To explore the mechanism of miR-671-5p regulating the proliferation,migration and invasion of nonsmall cell lung cancer cell lines.Methods The mRNA expression levels of miR-671-5p and Kruppel-like zinc finger transcription factor 6gene(KLF6)in human normal lung epithelial cell lines(BEAS-2B)and non-small cell lung cancer cell lines(A549and H460)were detected by RT-qPCR.The miR-671-5p mimic was transfected into A549and H460cells,and a negative control group was set.The expression of KLF6protein was detected by Western blotting.The proliferation,migration and invasion ability of non-small cell carcinoma cells were detected by CCK8,EDU and Transwell assays,respectively.Results Compared with BEAS-2Bcells,miR-671-5p was over-expressed in A549and H460cells,and the miR-671-5p expression in A549and H460cells was 1.646±0.026and 1.638±0.027,respectively(F=153.900,P<0.001).The KLF6mRNA expression in A549and H460cells down-regulated to 0.233±0.005and 0.382±0.003,respectively,and the difference was statistically significant(F=2 077.000,P<0.001).After transfection with miR-671-5p mimic,the proliferation,migration and invasion ability of A549and H460cells enhanced(all P<0.05),while the KLF6mRNA and protein expression levels were inhibited,and the KLF6protein levels in A549and H460cells decreased to0.252±0.011 (t=23.970,P<0.001)and 0.290±0.017 (t=16.140,P<0.001).Conclusion miR-671-5p may promote the proliferation,migration and invasion of non-small-cell lung cancer cell lines by regulating KLF6gene,and plays a regulatory role in the occurrence and progression of non-small-cell lung cancer.

关 键 词：miR-671-5p 非小细胞肺癌 增殖 迁移侵袭 KLF6基因 

分 类 号：R734.2[医药卫生—肿瘤]