陈蓉, 周宇, 赵富锋, 马铭. miR-1290通过抑制干扰素调节因子-2调控非小细胞肺癌的增殖[J]. 中国肿瘤临床, 2018, 45(16): 821-826. DOI: 10.3969/j.issn.1000-8179.2018.16.328
引用本文: 陈蓉, 周宇, 赵富锋, 马铭. miR-1290通过抑制干扰素调节因子-2调控非小细胞肺癌的增殖[J]. 中国肿瘤临床, 2018, 45(16): 821-826. DOI: 10.3969/j.issn.1000-8179.2018.16.328
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Chen Rong, Zhou Yu, Zhao Fufeng, Ma Ming. MiR-1290 regulates proliferation of non-small cell lung cancer by targeting interferon regulatory factor 2[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2018, 45(16): 821-826. DOI: 10.3969/j.issn.1000-8179.2018.16.328
Citation: Chen Rong, Zhou Yu, Zhao Fufeng, Ma Ming. MiR-1290 regulates proliferation of non-small cell lung cancer by targeting interferon regulatory factor 2[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2018, 45(16): 821-826. DOI: 10.3969/j.issn.1000-8179.2018.16.328
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miR-1290通过抑制干扰素调节因子-2调控非小细胞肺癌的增殖

MiR-1290 regulates proliferation of non-small cell lung cancer by targeting interferon regulatory factor 2

    摘要
  • 摘要:
      目的  探讨微小RNA-1290（miR-1290）在非小细胞肺癌（non-small cell lung cancer，NSCLC）组织中的表达及其相关调控机制。
      方法  采用实时荧光定量PCR检测成都市双流区第一人民医院2014年6月至2017年6月41例NSCLC患者癌组织、癌旁组织以及NSCLC细胞株A549、H460及正常支气管上皮细胞BEAS-2B中miR-1290表达；qPCR、Western blot检测癌组织、癌旁组织IRF2 mRNA和蛋白表达；将A549和H460细胞分为miR-1290 mimic组（转染miR-1290 mimic）、miR-1290 inhibit组（转染miR-1290 inhibit）和miR- 1290 NC组（不转染）。分别于转染24、48、72、96 h后采用CCK-8法检测细胞增殖能力，平板克隆形成实验检测细胞克隆形成数，Transwell小室检测细胞侵袭数，双荧光素酶报告基因实验检测miR-1290与干扰素调节因子-2（interferon regulatory factor，IRF2）3′-UTR结合情况，qPCR检测不同miR-1290表达水平的A549和H460细胞IRF2 mRNA表达。
      结果  癌组织miR-1290相对表达量明显高于癌旁组织，IRF2 mRNA和蛋白相对表达量明显低于癌旁组织，差异具有统计学意义（P < 0.05）；NSCLC组织miR-1290表达与IRF2 mRNA、蛋白表达呈显著负相关（P < 0.05）。NSCLC患者中，淋巴结转移者miR-1290表达明显高于无淋巴结转移者，Ⅲ/Ⅳ期患者miR-1290表达明显高于Ⅰ/Ⅱ期，差异具有统计学意义（P < 0.05）。转染72、96 h后，A549和H460细胞miR-1290 mimic组增殖能力明显高于miR- 1290 NC组，miR-1290 inhibit组增殖能力明显低于miR-1290 NC组，差异具有统计学意义（P < 0.05）。A549和H460细胞中miR-1290 mimic组细胞克隆数和侵袭细胞数明显高于miR-1290 NC组，miR-1290 inhibit组克隆数和侵袭细胞数明显低于miR-1290 NC组，差异具有统计学意义（P < 0.05）。双荧光素酶报告基因实验显示miR-1290能与IRF2 3′-UTR结合，明显降低荧光值（P < 0.05）。A549和H460细胞中，miR-1290 mimic组IRF2相对表达量明显低于miR-1290 NC组，miR-1290 inhibit组IRF2相对表达量明显高于miR-1290 NC组，差异具有统计学意义（P < 0.05）。
      结论  miR-1290在NSCLC组织和细胞中高表达，miR-1290可能通过与IRF2结合，促进肿瘤细胞的增殖和侵袭。

     

    Abstract:
      Objective  To investigate the expression of miR-1290 in non-small cell lung cancer (NSCLC) tissues and its regulation mechanism.
      Methods  This study evaluated 41 NSCLC patients enrolled in The First Hospital of Shuangliu from June 2014 to 2017. The expression of miR-1290 in NSCLC tissues, adjacent normal tissues, two NSCLC cell lines (A549 and H460), and one normal bronchial epithelial cell line (BEAS-2B) was evaluated by real-time quantitative PCR (qPCR). qPCR and Western blot were conducted to detect the expression of IRF2 mRNA and protein in cancer tissues and adjacent normal tissues. A549 and H460 cells were divided into three groups: miR- 1290 mimic (transfected with miR-1290 mimic), miR-1290 inhibit (transfected with miR-1290 inhibitor), and miR-1290 NC (blank control). At 24, 48, 72, and 96 h after transfection, cell proliferation, colony formation, and invasion ability were detected by CCK-8, colony forming, and Transwell assays, respectively. Luciferase activity was detected and analyzed with a dual-luciferase reporter system. IRF2 mRNA expression in A549 and H460 cells with different miR-1290 expression was detected by qPCR.
      Results  MiR-1290 expression levels were remarkably upregulated in NSCLC tissues compared with adjacent normal tissues. In addition, compared to adjacent normal tissues, the mRNA and protein expression of IRF2 was significantly downregulated in NSCLC tissues (all P < 0.05). There was an obvious negative correlation between miR-1290 and IRF mRNA and protein expression in NSCLC tissues (P < 0.05). Moreover, higher miR-1290 expression levels were positively associated with lymph node metastasis and an advanced tumor stage (Ⅲ/Ⅳ stage) (all P < 0.05). Functional assays showed that upregulated miR-1290 expression in NSCLC cells enhanced cell proliferation, cell colony formation, and invasion capacities in vitro, while downregulated miR-1290 expression had the opposite effects. Luciferase reporter gene assays revealed that IRF2 is a direct target gene of miR-1290. In both A549 and H460 cells, upregulated miR-1290 expression could depress the expression of IRF2 mRNA and protein; these effects were reversed by downregulating miR-1290.
      Conclusions  MiR-1290 is upregulated in NSCLC tissues and cells and enhances tumor proliferation and invasion by targeting IRF2 in NSCLC.

     

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