miR-933调控KLF6基因影响非小细胞肺癌的作用研究  

miR-933 inhibits proliferation, migration and invasion of lung cancer cell lines by regulation of KLF6 gene

作　　者：李海洲 张艳炜 许英杰 杨门 张磊[1] 韩京军[1] LI Haizhou;ZHANG Yanwei;XU Yingjie;YANG Men;ZHANG Lei;HAN Jingjun(Department of Thoracic and Cardiovascular Surgery,The Eighth Affiliated Hospital of Sun Yat-sen University,Shenzhen 518000,Guangdong Province,China;Department of Immune-Planning,Shenzhen Center for Disease Control and Prevention,Shenzhen 518055,Guangdong Province,China)

机构地区：[1]中山大学附属第八医院心胸外科,广东深圳518000 [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：《中国癌症杂志》2021年第7期581-588,共8页China Oncology

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

摘　　要：背景与目的:miRNA被认为参与肿瘤的发生、发展过程,但miRNA与肺癌的关系仍不完全清楚,探讨miR-933调控Kruppel样锌指转录因子6(Kruppel-like zinc finger transcription factor 6,KLF6)对肺癌细胞系增殖、迁移侵袭和诱导凋亡的影响。方法:采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQPCR)检测正常支气管上皮细胞BEAS-2B细胞、肺癌细胞系A549、H460细胞中miR-933的表达。采用RTFQ-PCR和蛋白质印迹法(Western blot)检测KLF6 mRNA表达和蛋白水平。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)和EdU法检测细胞增殖,采用transwell小室实验检测细胞迁移和侵袭,采用AnnexinⅤ-异硫氰酸荧光素(fluorescein isothiocyanate,FITC)/碘化丙啶(propidiumiodide,PI)染色法检测细胞凋亡。结果:肺癌细胞系转染miR-933mimic组KLF6mRNA表达水平明显上调(P<0.05)。与阴性对照组相比,高表达miR-933能增加A549、H460细胞KLF6蛋白的相对表达水平(P<0.05)。过表达miR-933可抑制A549、H460细胞的增殖、迁移和侵袭能力,差异均有统计意义(P<0.05)。转染miR-933mimc后,A549和H460细胞的凋亡率均显著高于各阴性对照组(P<0.001)。结论:miR-933通过调控KLF6基因的表达,诱导肺癌细胞的凋亡,抑制肺癌细胞的增殖,降低肺癌细胞的迁移和侵袭能力,影响肺癌的发生、发展。Background and purpose:miRNA is supposed to be involved in the occurrence and progression of tumors. However, studies are still inadequate. This study aimed to investigate whether miR-933 can inhibit cell proliferation, migration and invasion and induce apoptosis of lung cancer cell lines A549 and H460 by regulating Kruppel-like zinc finger transcription factor 6(KLF6). Methods:Expression of miR-933 in lung cancer cell lines(A549, H460) and bronchial epithelial cells(BEAS-2 B) was detected using real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR). Lung cancer cells were transfected with miR-933 mimic and mimic NC, respectively. RTFQ-PCR and Western blot were performed to detect expression of KLF6 in A549 and H460. Cell proliferation was detected by cell counting kit-8(CCK-8) assay and EdU assay, migration and invasion were detected by transwell assay, and apoptosis was detected by Annexin Ⅴ-fluorescein isothiocyanate(FITC)/propidium iodide(PI) staining. Results:Results of RTFQ-PCR showed that the expression of KLF6 in lung cancer cell lines transfected with miR-933 mimic was obviously upregulated(P<0.05). Western blot assay displayed that expression of KLF6 protein increased in A549 and H460 transfected with miR-933 mimic separately(P<0.05). Compared with negative control, the abilities of cell proliferation, migration and invasion in both A549 and H460 cells were significantly inhibited by overexpression of miR-933(P<0.05). Annexin Ⅴ-FITC/PI staining results showed that apoptotic rates were 48.3%±1.0% and 6.1%±0.2% respectively in A549 and H460 cells after transfection of miR-933 mimic, compared with negative control(37.6%±0.9%, 2.7%±0.01%)(P<0.001). Conclusion:By regulating KLF6, miR-933 induces apoptosis, inhibits cell proliferation, reduces abilities of migration and invasion of lung cancer cells and delays the occurrence and development of lung cancer.

关 键 词：MIRNA 非小细胞肺癌 细胞增殖 细胞凋亡 Kruppel样锌指转录因子6 

分 类 号：R734.2[医药卫生—肿瘤]