长链非编码RNA CCAT1通过促进上皮间充质转化对口腔鳞状细胞癌细胞侵袭和迁移能力的影响  

Effect of long non-coding RNA CCAT1 on abilities of invasion and migration in oral squamous cell carcinoma cells through promoting epithelial-to-mesenchy-mal transition

作　　者：陈敏[1] 张楠[1] 周政[1] CHEN Min;ZHANG Nan;ZHOU Zheng(Department of Stomatology,the First Affiliated Hospital of the Medical College,Shihezi University,Shihezi 832008,China)

机构地区：[1]石河子大学医学院第一附属医院口腔科

出　　处：《中国免疫学杂志》2019年第15期1845-1851,共7页Chinese Journal of Immunology

基　　金：2013年度科技惠民计划项目(2013GS650104)

摘　　要：目的:探究长链非编码RNA(LncRNA)结肠癌相关转录因子1(CCAT1)对口腔鳞状细胞癌SCC-9细胞侵袭、迁移及上皮间充质转化的影响及机制。方法:利用RT-PCR检测CCAT1在癌旁组织、口腔鳞状细胞癌组织、口腔角质细胞系hNOK、口腔鳞状上皮细胞癌细胞系SCC-4、SCC-9、SCC-25 和舌鳞状细胞癌Tca83细胞系的表达情况。进而用sh-CCAT1转染SCC9细胞,用CCK8检测细胞增殖情况,Hoechst染色检测细胞凋亡情况,划痕检测细胞迁移能力,Transwell检测细胞侵袭能力,免疫印迹检测增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、血管内皮生长因子(VEGF)、钙依赖性黏附蛋白E(E-cadherin)、钙依赖性黏附蛋白N(N-cadherin)、Snail、β-连环素(β-catenin)、T细胞因子4(TCF-4)、细胞周期蛋白D1(Cyclin-D1)的表达水平。结果: CCTA1的mRNA水平在口腔鳞状细胞癌组织及细胞中显著升高,且其在SCC-9中的mRNA水平高于其他口腔鳞状细胞癌细胞系,故选用SCC-9进行后续实验。sh-CCAT1可显著下调SCC-9细胞中CCAT1的mRNA水平,降低SCC-9细胞增殖速率及PCNA的表达,提高SCC-9凋亡细胞百分比,并降低SCC-9细胞划痕闭合率,减少SCC-9侵袭细胞数及MMP-2和VEGF的表达。同时,sh-CCAT1可显著提高E-cadherin的表达,降低N-cadherin和 Snail的表达。此外,sh-CCAT1显著降低β-catenin、TCF4和Cyclin-D1的表达。结论: CCAT1可通过促进上皮间充质转化增强口腔鳞状细胞癌SCC-9细胞侵袭和迁移能力。Objective: To investigate the effect of long non-coding RNA(LncRNA)Colon cancer associated transcript 1(CCAT1)on the abilities of cell invasion,migration and epithelial-to-mesenchymal transition in oral squamous cell carcinoma(SCC)cell line SCC-9. Methods: The mRNA levels of CCAT1 in normal tissue,oral squamous cell carcinoma tissue,human normal oral keratinocytes(hNOK),oral squamous cell carcinoma cell line SCC-4,SCC-9,SCC-25,tongue squamous cell line(Tca83)were measured by reverse transcription-PCR.SCC9 was selected to be transfected with sh-CCAT1.The cell proliferation and apoptosis were evaluated with cell counting kit-8 and hoechst staining.Wound healing assay was performed to evaluate cell migration ability,Transwell was performed to evaluate cell invasion ability and Western blot was performed to evaluate the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vascular endothelial growth factor(VEGF),calcium-dependent adhesion protein E(E-cadherin),N-cadherin,Snail,β-catenin,T cell factor 4(TCF-4)and Cyclin-D1. Results: The mRNA levels of CCTA1 were increased significantly in oral squamous cell carcinoma tissue and cells,and expressed highest in SCC-9.Thus SCC-9 was chosen for further study.Sh-CCAT1 could observably reduce the mRNA levels of CCTA1 in SCC-9 cell,decrease the rate of cell proliferation and the expression of PCNA,and enhance the percentage of apoptotic cell.And it lowered the rate of cell migration,the number of invasive cell and the expression of MMP-2 and VEGF.Meanwhile,sh-CCAT1 could significantly improve the expression of E-cadherin but lower the expression of N-cadherin and Snail.Furthermore,sh-CCAT1 decreased the expression of β-catenin,TCF4 and Cyclin-D1. Conclusion: CCAT1 could enhance the abilities of invasion and migration in oral squamous cell carcinoma SCC-9 cells through promoting epithelial-to-mesenchymal transition.

关 键 词：CCAT1 上皮间充质转化 口腔鳞状细胞癌 侵袭 迁移 

分 类 号：R739.85[医药卫生—肿瘤]