HIF-1α诱导结肠癌HCTT116细胞EMT和侵袭转移的作用机制研究  

The role and mechanism of HIF-1αinducing EMT and invasion and metastasis of colon cancer HCTT116 cells by regulating transcription factor Snail1

作　　者：李蓉蓉[1] 殷先利[1] 刘振洋[1] 杨晓琳[1] LI Rong-rong;YIN Xian-li;LIU Zhen-yang;YANG Xiao-lin(Department of Gastroenterology,Hunan Cancer Hospital,Hunan Changshu 410006)

机构地区：[1]湖南省肿瘤医院消化泌尿内科,410006

出　　处：《现代消化及介入诊疗》2020年第7期893-898,共6页Modern Interventional Diagnosis and Treatment in Gastroenterology

摘　　要：目的探究缺氧诱导因子-1α(HIF-1α)对结肠癌HCT116细胞侵袭转移能力以及对锌指转录因子Snail1和上皮间充质转化(EMT)标志蛋白表达的影响,阐明HIF-1α在结肠癌细胞侵袭转移中的作用及机制。方法选取人结肠癌HCT116细胞为研究对象,分别采用慢病毒细胞转染和siRNA技术、qRT-PCR、Western blot以及Transwell迁移和侵袭实验观察HIF-1α对结肠癌HCT116细胞侵袭转移能力以及对锌指转录因子Snail1和EMT标志分子mRNA及蛋白表达水平的影响。结果qRTPCR和Western blot结果显示,HIF-1α过表达明显上调结肠癌HCT116细胞中Snail1及间质细胞标志蛋白N-cadherin、EMT相关转录因子Slug和Smad3的表达,同时抑制上皮样标志蛋白E-cadherin的表达(P<0.05),而HIF-1α敲减能够逆转该促进作用。Transwell迁移和侵袭实验结果表明,HIF-1α过表达明显促进结肠癌HCT116细胞的迁移和侵袭,而敲减HIF-1α能够明显抑制HCT116细胞的迁移和侵袭能力(P<0.05);此外,采用KEGG信号通路富集分析以及Western blot结果表明,HIF-1α过表达显著激活结肠癌HCT116细胞中PI3K/PKB/GSK-3β/Snail1信号通路,进而下调E-cadherin的表达,发挥促结肠癌细胞EMT和转移作用,上述作用能够被PI3K/PKB/GSK-3β/Snail1信号通路特异性抑制剂Wortmannin逆转,同时HIF-1α敲减HCT116细胞中该信号通路活性明显降低(P<0.05)。结论HIF-1α通过激活PI3K/PKB/GSK-3β/Snail1信号通路促进结肠癌HCT116细胞EMT和侵袭转移,对临床结肠癌患者的诊断和靶向治疗具有重要指导价值,有望成为临床抗结肠癌转移治疗的潜在作用靶点。Objective To investigate the effect of hypoxia-induced factor-1α(HIF-1α)on invasion and metastasis of colon cancer HCT116 cells and the expression of zinc finger transcription factors Snail1 and EMT markers,and to clarify the role and mechanism of hif-1 a in invasion and metastasis of colon cancer cells.Methods Human colon cancer HCT116 cells were selected as the research subjects.Lentiviral cell transfection and siRNA techniques,qRT-PCR,Western blot,and transwell-migration and invasion assays were used to observe the effect of HIF-1αon the invasion and metastasis of colon cancer HCT116 cells and the mRNA and protein expression of zinc finger transcription factor Snail1 and EMT markers,respectively.Results qRT-PCR and Western blot showed that HIF-1αoverexpression significantly up-regulated the expression of Snail1 and mesenchymal-like marker N-cadherin,EMTrelated transcription factors Slug and Smad3 in colon cancer HCT116 cells,and inhibited the expression of epithelial-like marker Ecadherin(P<0.05),which can be reversed by HIF-1αknockdown.Transwell-migration and invasion assay results showed that HIF-1αoverexpression significantly promoted the migration and invasion of colon cancer HCT116 cells,and knockdown of HIF-1αsignificantly inhibited the migration and invasion of HCT116 cells(P<0.05).In addition,KEGG signaling pathway enrichment analysis and Western blot confirmed that HIF-1αoverexpression significantly activates the PI3 K/PKB/GSK-3β/Snail1 signaling pathway in colon cancer HCT116 cells,thereby down-regulating the expression of E-cadherin and exerting EMT and metastatic effects on colon cancer cells,which can be affected by PI3 K/PKB/GSK-3β/Snail1 signaling pathway specific inhibitor Wortmannin reversed,and HIF-1αknockdown HCT116 cells significantly reduced the activity of this signaling pathway(P<0.05).Conclusion HIF-1αpromotes EMT and invasion and metastasis of colon cancer HCT116 cells by activating PI3 K/PKB/GSK-3β/Snail1 signaling pathway,which has important guiding value for diagnosis a

关 键 词：HIF-1Α 皮肤鳞状细胞癌 上皮间质转化(EMT) 迁移 侵袭 

分 类 号：R574[医药卫生—消化系统]