miR-182调控RECK信号通路在非小细胞肺癌进展中的作用

Effect of mi R-182 in non small cell lung cancer via regulating the RECK signaling pathway

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 explore the effect of mi R-182 in non small cell lung cancer（NSCLC） and its mechanism. Methods The expression level and difference of mi R-182 were detected by q RT-PCR in NSCLC cell line and matched tissues of 30 patients suffering from NSCLC. Cell proliferation, invasion and metastasis of A549 and H1299 via mi R-182 was detected by mi R-182 mimics. The predicted target gene（RECK） of mi R-182 was identified via luciferase activation assay. The mRNA or protein expression of RECK after mi R-182 mimic or inhibitor transfection was detected by q RT-PCR or Western blotting, respectively. Results Expression of mi R-182 was significantly increased in NSCLC tissues and cell lines compared with that in the adjacent tissues（P〈0.01）, and it was correlated with the metastasis of NSCLC. The cell proliferation, invasion, and migration capacity was increased after mi R-182 mimics transfection. Dual luciferase assay showed that mi R-182 directly regulated the RECK translation level. RECK expression level was decreased after mi R-182 mimics transfection in the A549 cells, while increased after mi R-182 inhibitor transfection. Conclusion mi R-182 promoted tumor growth and metastasis of NSCLC by down-regulating the RECK expression.