Guo Huihui, Zhang Xilin, Chen Qiuqiang, Bao Ying, Dong Chaohui, Wang Xiang
Key Laboratory for Translational Medicine, First Affiliated Hospital, Huzhou UniversityHuzhou 313000, Zhejiang, China.
Am J Transl Res. 2018 Jan 15;10(1):224-234. eCollection 2018.
miR-132, a microRNA, has been reported to be down-regulated in several human cancers and is related with tumor progression; however, its function in non-small cell lung cancer (NSCLC) progression remains unclear. This study aimed to investigate the putative role of miR-132 in the metastasis of NSCLC. We determined the function of miR-132 in the migration and invasion of a NSCLC cell line using a miR-132 inhibitor and mimic. Our results showed overexpression of miR-132 significantly inhibited the migration and invasion of NSCLC cells . We then identified USP9X as a potential target of miR-132, and demonstrated miR-132 could regulate the expression of USP9X at both the mRNA and protein level. miR-132 could directly bind to the 3' untranslated region (3'-UTR) of USP9X. Inhibition of USP9X by its inhibitor WP1130 reduced the migration and invasion of NSCLC cells. Furthermore, USP9X inhibition also reversed the increased migration and invasion mediated by miR-132 inhibition. We found USP9X inhibition up-regulated expression of the epithelial-mesenchymal transition (EMT) marker E-cadherin, but down-regulated vimentin expression. A similar effect was seen with miR-132 overexpression, while the opposite effect occurred with miR-132 knockdown. USP9X inhibition reversed the miR-132 inhibitor-induced vimentin up-regulation and E-cadherin down-regulation. Taken together, these results indicate miR-132 prohibits the migration and invasion of NSCLC cells via targeting USP9X-induced EMT. Our data provides further evidence for the critical role of miR-132 and USP9X in regulating cell invasion and migration of NSCLC.
微小RNA miR-132已被报道在多种人类癌症中表达下调，并与肿瘤进展相关；然而，其在非小细胞肺癌（NSCLC）进展中的作用仍不清楚。本研究旨在探讨miR-132在NSCLC转移中的假定作用。我们使用miR-132抑制剂和模拟物确定了miR-132在NSCLC细胞系迁移和侵袭中的功能。我们的结果表明，miR-132的过表达显著抑制了NSCLC细胞的迁移和侵袭。然后，我们确定泛素特异性蛋白酶9X（USP9X）是miR-132的潜在靶点，并证明miR-132可以在mRNA和蛋白质水平上调节USP9X的表达。miR-132可以直接与USP9X的3'非翻译区（3'-UTR）结合。其抑制剂WP1130对USP9X的抑制作用降低了NSCLC细胞的迁移和侵袭。此外，USP9X抑制还逆转了miR-132抑制介导的迁移和侵袭增加。我们发现USP9X抑制上调了上皮-间质转化（EMT）标志物E-钙黏蛋白的表达，但下调了波形蛋白的表达。miR-132过表达也有类似效果，而miR-132敲低则产生相反效果。USP9X抑制逆转了miR-132抑制剂诱导的波形蛋白上调和E-钙黏蛋白下调。综上所述，这些结果表明miR-132通过靶向USP9X诱导的EMT来抑制NSCLC细胞的迁移和侵袭。我们的数据为miR-132和USP9X在调节NSCLC细胞侵袭和迁移中的关键作用提供了进一步证据。
