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目的探讨低氧对分化型甲状腺癌KAT、WRO细胞上皮间质转化(Epithelial-mesenchymal transition,EMT)的影响。方法以氯化钴(CoCl2)模拟细胞低氧微环境,采用MTT法检测细胞的增殖活力,筛选CoCl2的最适作用浓度;倒置显微镜观察低氧下细胞的形态特征;Western blot检测低氧0、3、6、12 h后KAT和WRO细胞中HIF-1a蛋白的表达及低氧6 h后KAT和WRO细胞上皮细胞钙黏蛋白(E-cadherin)和间质细胞波形蛋白(Vimentin)的表达。结果 CoCl2最适作用浓度为150μmol/L,低氧使KAT和WRO细胞逐渐获得了间质细胞的形态特征。低氧6 h后,KAT和WRO细胞中HIF-1a蛋白的表达达峰值,与低氧0 h组相比,差异有统计学意义(P<0.01)。低氧组KAT细胞中E-cadherin蛋白的表达水平明显低于正常组(P<0.01),Vimentin蛋白的表达水平与正常组相比,差异无统计学意义(P>0.05);低氧组WRO细胞中E-cadherin蛋白的表达水平明显低于正常组(P<0.01),Vimentin蛋白的表达水平明显高于正常组(P<0.01)。结论 HIF-1a可能通过下调E-cadherin、上调Vimentin而促进分化型甲状腺癌细胞KAT和WRO发生上皮间质转化,使其获得侵袭、转移潜能。
Objective To investigate the effect of hypoxia on Epithelial-mesenchymal transition (EMT) in differentiated thyroid cancer KAT and WRO. Methods The cell hypoxia microenvironment was simulated by cobalt chloride (CoCl 2), the cell viability was detected by MTT assay and the optimal concentration of CoCl 2 was screened. The morphology of hypoxic cells was observed by inverted microscope. The expression of HIF-1a protein in KAT and WRO cells and the expression of E-cadherin and Vimentin in KAT and WRO cells after 3, 6, and 12 h of hypoxia were observed. Results The optimum concentration of CoCl2 was 150 μmol / L. Hypoxia led to the progressive morphological characteristics of stromal cells in KAT and WRO cells. The expression of HIF-1a protein in KAT and WRO cells peaked at 6 h after hypoxia, which was significantly different from that in hypoxia group at 0 h (P <0.01). The expression of E-cadherin protein in KAT cells in hypoxia group was significantly lower than that in normal group (P <0.01), and the expression level of Vimentin protein in normal hypoxia group was not significantly different from that in normal group (P> 0.05) The expression of E-cadherin protein in cells was significantly lower than that in normal group (P <0.01), and the expression of Vimentin protein was significantly higher than that in normal group (P <0.01). Conclusions HIF-1a may promote epithelial-mesenchymal transition of differentiated thyroid carcinoma cells by down-regulating E-cadherin and up-regulating Vimentin so as to obtain invasion and metastasis potential.