miR-670-5p对肺癌细胞增殖、迁移、侵袭的影响

目的: 探讨miR-670-5p对肺癌细胞增殖、迁移和侵袭的影响,分析其调控WW结构域氧化还原酶基因(WWOX)的机制。方法: 收集2016年1月至2017年10月收治的28例肺癌组织和对应癌旁组织,实时荧光定量PCR(RT-qPCR)检测肺癌组织、癌旁组织中miR-670-5p的表达水平。将肺癌细胞A549分为anti-miR-NC组(转染anti-miR-NC)、anti-miR-670-5p组(转染anti-miR-670-5p)、anti-miR-670-5p+si-NC组(转染anti-miR-670-5p与si-NC)、anti-miR-670-5p+si-WWOX组(转染anti-miR-670-5p与si-WWOX)。转染48 h后,RT-qPCR或蛋白质印记(Western blot)检测转染效果。细胞计数试剂盒(CCK-8)检测细胞活力;Transwell实验检测细胞迁移和侵袭能力;Western blot检测P21、上皮细胞钙粘蛋白(E-cadherin)和基质金属蛋白酶2(MMP-2)蛋白的表达水平。双荧光素酶报告基因实验和Western blot验证miR-670-5p和WWOX的靶向关系。结果: 肺癌组织中miR-670-5p的表达水平较癌旁组织显著升高(P＜0.05)。抑制miR-670-5p可抑制MMP-2蛋白表达(P＜0.05),促进P21和E-cadherin表达(P＜0.05),抑制A549细胞增殖、迁移和侵袭(P＜0.05)。WWOX是miR-670-5p的靶基因,miR-670-5p负调控WWOX表达。抑制WWOX可部分逆转anti-miR-670-5p对A549细胞增殖、迁移和侵袭的影响(P＜0.05)。结论: miR-670-5p通过靶向WWOX能够促进肺癌细胞增殖、迁移、侵袭。

Effects of miR-670-5p on proliferation,migration and invasion of lung cancer cells

Objective: To investigate the effects of miR-670-5p on the proliferation, migration and invasion of lung cancer cells, further to analyze its mechanisms of regulating WW domain oxidoreductase gene (WWOX). Methods: From January 2016 to October 2017, 28 cases of lung cancer tissues and corresponding adjacent tissues were collected. And the expressions of miR-670-5p in lung cancer tissues and adjacent tissues were detected by RT-qPCR. Lung cancer cells A549 were divided into anti-miR-NC group (transfected with anti-miR-NC), anti-miR-670-5p group (transfected with anti-miR-670-5p), and anti-miR-670-5p+ si-NC group (transfected with anti-miR-670-5p and si-NC), anti-miR-670-5p+si-WWOX group (transfected with anti-miR-670-5p and si-WWOX). After 48 hours of transfection, RT-qPCR or Western Blot were used to detect the transfection effects. Cell viability was detected by using CCK-8 method; cell migration and invasion were detected by using Transwell assay; Western blot was used to analyze the expression levels of P21, E-cadherin and MMP-2 protein. The dual luciferase reporter gene assay and Western blot were applied to verify the targeting relationship between miR-670-5p and WWOX. Results: The expression level of miR-670-5p in lung cancer tissues was significantly higher than that in adjacent tissues (P＜0.05). Inhibition of miR-670-5p decreased MMP-2 protein expression (P＜0.05), increased P21 and E-cadherin expressions (P＜0.05), and inhibited proliferation, migration and invasion of A549 cells (P＜0.05). WWOX was a target gene of miR-670-5p, and miR-670-5p negatively regulated WWOX expression. Inhibition of WWOX partially reversed the effect of anti-miR-670-5p on proliferation, migration and invasion of A549 cells (P＜0.05). Conclusion: miR-670-5p promotes proliferation, migration and invasion of lung cancer cells by targeting WWOX.