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目的 :探讨低氧微环境诱导人舌鳞癌细胞上皮-间质转化以及HIF-1α在该过程中的作用。方法 :体外常氧(20%O_2)或低氧(1%O_2)状态下培养舌鳞癌SCC9、CAL27细胞48 h,Western免疫印迹和实时定量PCR检测上皮间质标记蛋白vimentin、fibronectin和E-cadherin的表达变化和HIF-1α的表达水平,Transwell小室检测细胞的迁移能力。通过小干扰RNA(HIF-1α-Si)转染舌鳞癌细胞,检测HIF-1α、vimentin、fibronectin与E-cadherin蛋白表达以及细胞迁移能力的变化。应用SPSS13.0软件包对数据进行独立样本t检验。结果:人舌鳞癌细胞SCC9、CAL27在低氧环境中培养48 h后,间质标志蛋白vimentin和fibronectin在蛋白和m RNA水平均显著升高,上皮标志蛋白E-cadherin表达降低,HIF-1α表达上调,细胞迁移能力显著增强。小干扰RNA(siRNA)转染舌鳞癌细胞SCC9、CAL27并于低氧下培养后,HIF-1α表达显著降低,vimentin和fibronectin表达也显著降低,E-cadherin表达升高,细胞迁移能力显著下降。结论:低氧通过上调HIF-1α表达而促进人舌鳞癌细胞发生上皮-间质转化及转移。
Objective: To investigate the effect of hypoxia microenvironment on the epithelial-mesenchymal transition of human tongue squamous cell carcinoma and the role of HIF-1α in this process. Methods: SCC9 and CAL27 cells were cultured under normoxia (20% O 2) or hypoxia (1% O 2) for 48 h in vitro. The expressions of vimentin, fibronectin and E-cadherin were detected by Western blot and real- cadherin expression and HIF-1αexpression levels, Transwell chamber detection of cell migration. The tongue squamous cell carcinoma cells were transfected with small interfering RNA (HIF-1α-Si) to detect the protein expression of HIF-1α, vimentin, fibronectin and E-cadherin as well as the cell migration ability. SPSS13.0 software package for independent sample t-test data. Results: The expressions of vimentin and fibronectin were significantly increased in SCC9 and CAL27 cells cultured in hypoxic environment for 48 h, the expressions of E-cadherin, HIF-1α Upregulation of expression, cell migration ability was significantly enhanced. Small interfering RNA (siRNA) transfected tongue squamous cell carcinoma SCC9, CAL27 and cultured under hypoxia, the expression of HIF-1α was significantly decreased, the expression of vimentin and fibronectin was also significantly decreased, the expression of E-cadherin, cell migration was significantly decreased . Conclusion: Hypoxia promotes epithelial-mesenchymal transition and metastasis of human tongue squamous cell carcinoma by up-regulating the expression of HIF-1α.