MALAT1通过VHL／β-catenin通路影响口腔鳞状细胞癌生长侵袭的实验研究  

Metastasis-associated lung adenocarcinoma transcript 1 modulates the growth and invasion of oral squamous cell carcinoma through the VHL/β-catenin pathway in vitro

作　　者：王佳鑫[1] 段远胜[1] 岳恺[1] 司海山 董梦丽[1] 周梦倩[1] 王旭东[1] 

机构地区：[1]天津医科大学肿瘤医院颌面耳鼻喉肿瘤科国家肿瘤临床医学研究中心天津市肿瘤防治重点实验室,天津市恶性肿瘤临床医学研究中心,天津市300060

出　　处：《中国肿瘤临床》2017年第9期418-423,共6页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金项目(编号:81672684);天津医科大学肿瘤医院科研基金项目(编号:1601)资助~~

摘　　要：目的:探讨肺腺癌转移相关转录因子1(metastasis-associated lung adenocarcinoma transcript 1,MALAT1)对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)生长侵袭的影响及其作用机制。方法:使用小干扰RNA(small interfering RNA,siRNA)敲低MALAT1在人OSCC细胞系SCC25、UM1中的表达;CCK-8法、流式细胞术检测敲低MALAT1表达后OSCC增殖能力、细胞周期及细胞凋亡的变化;细胞迁移实验及Transwell实验检测细胞运动及侵袭能力的变化;蛋白质印迹法检测细胞周期及凋亡相关蛋白、上皮间质转化(epithelial-mesenchymal transition,EMT)及侵袭相关蛋白、VHL及β-catenin等蛋白表达变化。结果:敲低OSCC中MALAT1表达后,细胞增殖能力显著下降,细胞周期出现G1/S期阻滞,周期相关蛋白cyclin D1表达减少,P21表达增多;细胞凋亡增多,凋亡相关蛋白cleaved caspase-3、Bax表达增多;且EMT相关蛋白N-cadherin、vimentin表达减少,E-cadherin表达增多,侵袭相关蛋白MMP-2、MMP-9蛋白表达减少,细胞迁移、侵袭能力下降;同时发现敲低MALAT1表达后VHL蛋白表达增多,β-catenin蛋白表达减少。结论:MALAT1是OSCC生长侵袭的重要调节因素,可能通过VHL/β-catenin通路发挥调控作用。Objective： To investigate the effect and mechanism of metastasis-associated lung adenocarcinoma transcript 1 （MALAT1） in modulating the growth and invasion of oral squamous cell carcinoma （OSCC）. Methods： MALAT1 siRNA was transfected into SCC25 and UM1 human OSCC cell lines. Cell counting kit-8 assay and flow cytometry were used to test the proliferation, cell cycle, and apop- tosis of the cells after infection by the MALAT1 siRNA. Cell invasive and migration ability were evaluated by Transwell assay and cell migration assay, respectively. The expression levels of the proteins VHL and 13-catenin, which regulate the cell cycle, apoptosis, epithelial- mesenchymal transition, invasion, and migration, were examined by Western blot assay. Results： After down regulation of the MALAT1 expression in the cells, the proliferation was inhibited, and G1/S arrest was triggered. The expression of cyclin D1 was down regulated and that of P21 was up regulated. Cell apoptosis increased, and the expression levels of Bax and cleaved caspase-3 were up regulated. Migration and invasion were attenuated. The expression levels of N-cadherin, vimentin, and MMP-2/9 were down regulated, and the expression of E-cadherin was up regulated in the cells after the knockdown of MALAT1. These findings show significant differences be- tween the transfected cells and negative-control cells. We found that VHL expression was up regulated and that of β-catenin was down regulated. Conclusion： MALAT1 is an important factor for the growth and invasion of OSCC. MALAT1 possibly modulates these process- es through the VHL/β-catenin signal pathway.

关 键 词：口腔鳞状细胞癌 肺腺癌转移相关转录因子1 VHL Β-CATENIN 

分 类 号：R739.8[医药卫生—肿瘤]