MicroRNA-9影响涎腺腺样囊性癌侵袭能力的分子机制研究  

Molecular Mechanism of MicroRNA-9 Influencing Invasion of Salivary Adenoid Cystic Carcinoma Cells

作　　者：徐川[1,2] 李亚楠 谯丹[1] 古欢 钟罗 吴键 于涛[1,2] XU Chuan;LI Ya-nan;QIAO Dan;GU Huan;ZHONG Luo;WU Jian;YU Tao(Department of Head and Neck,the Affiliated Cancer Hospital,School of Medicine,University of Electronic Science and Technology of China,Chengdu 610041,China;Department of Head and Neck,Sichuan Cancer Hospital &Institute,Chengdu 610041,China)

机构地区：[1]电子科技大学附属肿瘤医院头颈外科,四川成都610041 [2]四川省肿瘤医院头颈外科,四川成都610041

出　　处：《口腔医学研究》2019年第6期563-567,共5页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81602387);四川省科技计划重点研发项目(编号:2018SZ0154);四川省应用基础研究(编号:2019YJ0572)

摘　　要：目的:探讨miR-9对人涎腺腺样囊性癌细胞侵袭的影响及其相关分子机制。方法:检测ACC-2,ACC-M细胞中miR-9表达,构建慢病毒miR-9过表达载体,转染ACC-2,ACC-M肿瘤细胞,实时荧光定量PCR检测细胞中 miR-9的表达水平。Transwell方法检测转染miR-9前后ACC-M细胞侵袭转移能力的变化,Western 免疫印迹检测转染前后ACC-M细胞中基质金属蛋白酶家族蛋白(MMP)及数个肿瘤相关信号通路蛋白的表达情况,采用 SPSS 16.0 软件包对数据进行统计学分析。结果:相比于ACC-2,ACC-M细胞中出现了显著的miR-9的表达下调( P <0.01)。携带miR-9的慢病毒转染细胞后均显著提高了2株细胞中miR-9的表达并显著降低了ACC-M细胞的侵袭能力( P <0.01)。同时,转染miR-9后显著降低了MMP-2及MMP-9蛋白表达及AKT蛋白的磷酸化水平。结论:上调miR-9 的表达可显著抑制ACC-M细胞的侵袭能力,其机制可能与AKT信号通路介导的MMP-2,9蛋白的表达下调密切相关。Objective: To investigate the effect and regulatory mechanisms of microRNA-9 on invasion of human salivary adenoid cystic carcinoma (SACC) cells. Methods: The expressions of microRNA-9 (miR-9) from ACC-2 cells and ACC-M cells were analyzed using qRT-PCR after transfected with miR-9 lentivirus.Cell invasion was measured by transwell experiment after transfection.Western blot was used to detect the protein expression levels of matrix metalloproteinases family and tumor-related signaling pathways in SACC cell lines after transfection.SPSS 16.0 was used to analyze the data. Results: MiR-9 expression was remarkably down-regulated in the ACC-M cells in contrast to ACC-2 cells ( P <0.01).After 72h transfection,the expression of miR-9 was increased in both ACC cells ( P <0.01).MiR-9 overexpression triggered to a decreased cell invasion after transfection for ACC-M cells ( P <0.01).The expressions of MMP-2 and MMP-9 proteins were significantly decreased after transfection in the ACC-M cells,while AKT protein phosphorylation was significantly decreased. Conclusion: Upregulation of miR-9 can inhibit the invasion of SACC cells.The mechanism might relate to the down-regulation of MMP-2 and MMP-9 mediated by AKT signaling pathway.

关 键 词：腺样囊性癌 微小RNA 基质金属蛋白酶 AKT信号通路 

分 类 号：R739.8[医药卫生—肿瘤]