过表达miR-129-5p通过靶向MAPK1抑制宫颈癌HeLa细胞恶性生物行为

[摘要] 目的：探讨miR-129-5p 对宫颈癌HeLa细胞侵袭、迁移和EMT的作用及其机制。方法：选取宫颈癌HeLa细胞，利用生物信息学预测软件筛选miR-129-5p 的靶基因，双荧光素酶报告基因验证miR-129-5p 和MAPK1 的靶向关系。将miR-129-5p mimic、miR-129-5p inhibitor 和pcDNA-MAPK1 单独或联合转染到HeLa细胞，用qPCR检测HeLa细胞中miR-129-5p 和MAPK1 的表达水平，用Transwell、划痕愈合实验分别检测HeLa 细胞的侵袭、迁移能力，WB检测细胞中E-cadherin、N-cadherin、MAPK1、STAT3 和Bcl-xL的表达。构建裸鼠HeLa细胞皮下移植瘤模型，观察miR-129-5p 过表达对移植瘤生长的影响，WB检测移植瘤组织中EMT及MAPK1 通路相关蛋白的表达。结果：miR-129-5p 与MAPK1 在3’UTR区存在结合位点，过表达miR-129-5p 靶向抑制MAPK1（P<0.01）。与对照组相比，miR-129-5p mimic 组侵袭细胞数目减少（P<0.01），划痕愈合率降低（均P<0.01）；细胞中Ecadherin表达上调而N-cadherin、MAPK1、STAT3 和Bcl-xL 表达下调（均P<0.01）；共转染MAPK1 可逆转上述现象。成功建立裸鼠HeLa 细胞移植瘤模型，与对照组相比，miR-128-3p mimic 组肿瘤质量减轻（P<0.01）；瘤组织中E-cadherin 表达水平上调而N-cadherin、MAPK1、STAT3 和Bcl-xL 的表达下调（均P<0.01）。结论：过表达miR-129-5p 通过靶向MAPK1 抑制宫颈癌HeLa细胞的侵袭、迁移和EMT。

Over-expression of miR-129-5p inhibits malignant biological behaviors of cervical cancer HeLa cells by targeting MAPK1

[Abstract] Objective: To investigate the effects and mechanisms of miR-129-5p on invasion, migration and epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cells. Methods:Cervical cancer HeLa cells were selected. The target gene of miR-129-5p was screened by bioinformatics prediction software, and the targeting relationship between miR-129-5p and MAPK1 was verified by dual luciferase reporter gene assay. HeLa cells were transfected with miR-129-5p mimic, miR-129-5p inhibitor and pcDNA-MAPK1 alone or in combination.The expressions of miR-129-5p and MAPK1 in HeLa cells were detected by qPCR; the invasion and migration ability of HeLa cells were detected by Transwell and scratch-healing experiments, respectively; and the expressions of E-cadherin, N-cadherin,MAPK1, STAT3 and Bcl-xL were detected by WB. The subcutaneous xenograft model of HeLa cells in nude mice was constructed to observe the effect of miR-129-5p over-expression on the growth of transplanted tumors. The expressions of EMT and MAPK1 pathwayrelated proteins in transplanted tumor tissues were detected by WB. Results: miR-129-5p could bind with the 3''UTR region of MAPK1,and over-expression of miR-129-5p targetedly inhibited the expression of MAPK1 (P<0.01). Compared with the control group, the number of invasive cells in the miR-129-5p mimic group decreased (P<0.01), the scratch healing rate decreased (all P<0.01); The expression of E-cadherin was up-regulated, and the expressions of N-cadherin, MAPK1, STAT3 and Bcl-xL were down-regulated (all P<0.01), while co-transfection of MAPK1 reversed the above phenomenon.The nude mice HeLa cell xenograft model was successfully established.Compared with the control group, the tumor mass of the miR-128-3p mimic group was reduced; the expression of E-cadherin was up-regulated in tumor tissues, while the expressions of N-cadherin, MAPK1, STAT3 and Bcl-xL were down-regulated (all P<0.01).Conclusion: Over-expression of miR-129-5p inhibits invasion, migration and epithelial-mesenchymal transition of cervical cancer HeLa cells by targeting MAPK1.