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目的探讨miR-182在非小细胞肺癌(NSCLC)发展中的作用及其相关机制。方法采用实时定量PCR检测NSCLC细胞系以及30组配对的NSCLC患者癌组织和癌旁组织样本中miR-182的表达水平。在NSCLC细胞株A549和H1299中过表达miR-182后,测定细胞增殖活性及侵袭转移能力。利用荧光素酶活性实验鉴定miR-182的预测靶基因(RECK)。采用qRT-PCR和Western blotting分别检测转染miR-182类似物和miR-182抑制物后RECK mRNA和蛋白的表达水平。结果miR-182在NSCLC细胞系和癌组织中的表达水平明显高于癌旁组织,在伴淋巴结转移的肺癌患者癌组织中的表达水平明显高于不伴淋巴结转移者,差异有统计学意义(P<0.01)。过表达miR-182可增强NSCLC细胞株A549和H1299的增殖、侵袭和迁移能力。双荧光素酶实验结果表明,RECK的翻译水平受miR-182直接调控。qRT-PCR和Western blotting 检测结果表明,转染miR-182类似物后,RECK表达水平降低,转染miR-182抑制物后,RECK表达水平升高。结论miR-182可调控RECK的表达,促进NSCLC的进展。
Objective To investigate the role of miR-182 in the development of non-small cell lung cancer (NSCLC) and its related mechanisms. Methods Real-time quantitative PCR was used to detect the expression of miR-182 in NSCLC cell lines and in 30 paired NSCLC tissues. After overexpression of miR-182 in NSCLC cell lines A549 and H1299, cell proliferation activity and invasion and metastasis were determined. The luciferase activity assay was used to identify the predicted target gene (RECK) of miR-182. The expression of RECK mRNA and protein were detected by qRT-PCR and Western blotting respectively after transfected with miR-182 analogues and miR-182 inhibitor. Results The expression level of miR-182 in NSCLC cell lines and cancer tissues was significantly higher than that in adjacent non-cancerous tissues. The expression level of miR-182 in patients with lung cancer with lymph node metastasis was significantly higher than that without lymph node metastasis (the difference was statistically significant ( P <0.01). Overexpression of miR-182 enhanced the proliferation, invasion and migration of NSCLC cell lines A549 and H1299. Dual luciferase results show that the translation level of RECK is directly regulated by miR-182. The results of qRT-PCR and Western blotting showed that the expression level of RECK decreased after transfection of miR-182 analogs, and the expression of RECK increased after transfected with miR-182 inhibitor. Conclusion miR-182 regulates RECK expression and promotes the progression of NSCLC.