HPV诱导的miR-21-5p通过PI3K/AKT途径促进宫颈癌细胞的上皮间质转化和肿瘤进展

HPV-induced miR-21-5p promotes epithelial-mesenchymal transition and tumor progression in cervical cancer cells via the PI3K/AKT pathway

摘要

目的 探讨人乳头瘤病毒(HPV)诱导的miR-21-5p对宫颈癌细胞的上皮间质转化(EMT)和肿瘤进展的影响。方法 HPV(+)宫颈癌细胞分为mimic-NC组、miR-21-5p-mimic组、inhibitor-NC和miR-21-5p-inhibitor组。采用实时荧光定量聚合酶链式反应(qPCR)分析miR-21-5p在HPV(+)宫颈癌细胞中的表达,Transwell实验检测细胞迁移和侵袭情况,Western blot检测上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)和β-肌动蛋白(β-actin)的表达。结果 与HPV(-)C33A相比,HPV16(+)SiHa和HPV18(+)HeLa中miR-21-5p的表达显著增加(F=51.35,P<0.01)。Transwell实验结果显示,与mimic-NC组相比,miR-21-5p-mimic组细胞迁移和侵袭数量显著提高(t=6.73,5.64,P<0.01);而与inhibitor-NC组相比,miR-21-5p-inhibitor组细胞迁移和侵袭数量显著降低(t=17.32,15.46,P<0.01)。Western blot检测结果显示,与mimic-NC组相比,miR-21-5p-mimic组细胞中E-cadherin表达降低,N-cadherin和Vimentin表达升高;而与inhibitor-NC组相比,miR-21-5p-inhibitor组细胞中E-cadherin表达升高,N-cadherin和Vimentin表达降低。与mimic-NC组相比,miR-21-5p-mimic组细胞中p-PI3K和p-AKT显著提高;而与inhibitor-NC组相比,miR-21-5p-inhibitor组细胞中p-PI3K和p-AKT显著降低。结论 HPV诱导的miR-21-5p可能通过激活PI3K/AKT途径促进宫颈癌细胞的迁移、侵袭和上皮间质转化。

Abstract

Objective To investigate the effects of HPV-induced miR-21-5p on epithelial-mesenchymal transition (EMT) and tumour progression in cervical carcinoma cells. Methods HPV(+) cervical cancer cells were divided into the mimic-NC group, miR-21-5p-mimic group, inhibitor-NC group,and miR-21-5p-inhibitor group.Real-time fluorescence quantitative polymerase chain reaction (qPCR) was used to analyze the expression of miR-21-5p in HPV (+) cervical cancer cells,and Transwell assay was detected cell migration and invasion.The expression of E-cadherin,N-cadherin,Vimentin,phosphorylated phosphatidylinositol 3-kinase (p-PI3K),phosphatidylinositol 3-kinase (PI3K),protein kinase B (AKT),phosphorylated protein kinase B (p-AKT) proteins and β-actin were detected by Western blot. Results Compared with HPV(-) C33A cells,the expression of miR-21-5p was significantly increased in HPV16(+ ) SiHa and HPV18(+ ) HeLa cells (F =51.35,P <0.01).Transwell assay results demonstrated that,compared with the mimic-NC group,the miR-21-5p-mimic group exhibited significantly increased cell migration and invasion ( t=6.73 and 5.64,respectively P<0.01);whereas compared with the inhibitor-NC group,cell migration and invasion were markedly reduced in the miR-21-5p-inhibitor group (t=17.32 and 15.46,respectively P <0.01).Western blot analysis revealed that,compared with the mimic-NC group,miR-21-5p-mimic-treated cells exhibited reduced E-cadherin expression alongside increased N-cadherin and Vimentin expression.Conversely,compared with the inhibitor-NC group,miR-21-5p-inhibitor group cells demonstrated elevated E-cadherin expression alongside decreased N-cadherin and vimentin expression.Compared with the mimic-NC group,p-PI3K and p-AKT levels were significantly elevated in miR-21-5p-mimic group cells.Conversely,compared with the inhibitor-NC group,p-PI3K and p-AKT levels were markedly reduced in miR-21-5p-inhibitor group cells. Conclusion HPV-induced miR-21-5p may promote migration, invasion and epithelial-mesenchymal transition in cervical cancer cells by activating the PI3K/AKT pathway.

关键词

HPV / miR-21-5p / PI3K/AKT途径 / 宫颈癌 / 上皮间质转化

Key words

HPV / miR-21-5p / PI3K/AKT pathway / cervical cancer / epithelial-mesenchymal transition

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基金资助

内蒙古医科大学青年创新基金项目(YKD2018QNCX072)