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.
作者
李海洲
张艳炜
许英杰
杨门
张磊
韩京军
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)
出处
《中国癌症杂志》
CAS
CSCD
北大核心
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)。
