miR-7704对头颈鳞状细胞癌细胞生物学行为的影响

The effects of miR-7704 on the biological behavior of HNSCC cells

目的:探究过表达miR-7704对头颈鳞状细胞癌(HNSCC)细胞生物学行为的影响。方法:应用实时定量RT-PCR检测人角质形成细胞HOK和HNSCC细胞系HN4、HN6、HN13及CAL27中miR-7704表达水平。构建过表达载体miR-7704 mimic转染HN6、CAL27。使用CCK8、平板克隆形成、划痕愈合实验及Transwell侵袭实验检测过表达miR-7704对HN6、CAL27细胞增殖、迁移和侵袭等生物学行为的影响。Western blot检测HN6、CAL27细胞过表达miR-7704后上皮-间充质转化(EMT)相关蛋白及苏氨酸激酶(AKT)信号通路表达。结果:相较HOK,HNSCC细胞系内miR-7704表达量均明显上调(P<0.05)。HN6、CAL27细胞转染过表达载体miR-7704 mimic后,细胞增殖、迁移及侵袭能力增强。Western blot结果显示过表达miR-7704后HN6、CAL27细胞上皮性钙黏附蛋白(E-cadherin)及β-连环蛋白(β-catenin)表达水平下降(P<0.01),神经性钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)、Snail蛋白及Slug蛋白表达水平均上调(P<0.01),AKT及p-AKT蛋白表达水平均明显升高(P<0.01)。结论:miR-7704可能作为一种促癌因子,通过调控AKT信号通路诱导EMT,从而促进HNSCC细胞生物学行为进程。

Objective:To investigate the influence of miR-7704 expression on the biological behavior of head and neck squamous cell carcinoma(HNSCC)cells.Methods:The discrepant expression of miR-7704 were detected by quantitative real-time RT-PCR(qRT-PCR)among human oral epithelial cell line HOK and HNSCC cell lines HN4,HN6,HN13,and CAL27.HN6 and CAL27 cells were transfected with miR-7704 mimic.The effects of miR-7704 overexpression on proliferation,migration and invasion of HN6 and CAL27 cells were investigated by CCK8,plate colony formation,wound healing experiment and Transwell invasion experiment.The expression of epithelial mesenchymal transition(EMT)related proteins and serine/threonine kinase(AKT)signaling pathway were detected by Western blot after HN6 and CAL27 cells transfected with miR-7704 mimic and mimic NC.Results:Compared with HOK cells,the expression of miR-7704 in all HNSCC cell lines were significantly up-regulated(P<0.05).Meanwhile,the proliferation,migration and invasion ability were enhanced after HN6 and CAL27 cells transfected with miR-7704 mimic.Western blot showed that the expression of E-cadherin andβ-catenin were decreased after overexpression of miR-7704 in HN6 and CAL27 cells(P<0.01),while the up-regulated expression of Vimentin,Snail and Slug protein were observed(P<0.01).The expression of AKT and p-AKT were significantly elevated(P<0.01).Conclusions:miR-7704 may act as a pro-oncogenic factor and induce EMT by regulating the AKT signaling pathway to promote the biological behavior of HNSCC cells.