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目的探讨低氧诱导因子-1α(Hypoxia-inducible factor-1α,HIF-1α)和上皮间质转化(Epithelial-mesenchymaltransition,EMT)相关蛋白在甲状腺滤泡状癌(Follicular thyroid carcinoma,FTC)组织中的表达及临床意义。方法采用免疫组化SP法检测42份FTC组织中HIF-1α、锌指转录因子(Snail)、波形蛋白(Vimentin)和钙黏蛋白E(E-cadherin)的表达;MTT法检测不同浓度CoCl2对FTC WRO细胞增殖活力的影响;RT-PCR法检测不同浓度CoCl2对WRO细胞HIF-1α基因mRNA转录水平的影响;免疫荧光法观察150μmol/L CoCl2处理24 h的WRO细胞中HIF-1α蛋白的核定位;Western blot检测150μmol/LCoCl2处理不同时间对WRO细胞中HIF-1α、Snail、Vimentin和E-cadherin蛋白表达的影响;Transwell小室试验检测150μmol/LCoCl2处理24 h对WRO细胞中侵袭及迁移能力的影响。结果在42份FTC组织中,HIF-1α、Snail、Vimentin和E-cadherin蛋白的表达与多项临床指标相关,且4者表达显著相关(P<0.05);CoCl2可抑制WRO细胞的增殖活力;随着CoCl2浓度的增高,WRO细胞中HIF-1α基因mRNA的转录水平先增高然后趋于平稳;150μmol/L CoCl2处理24 h的WRO细胞内HIF-1α蛋白发生核转位;CoCl2处理的WRO细胞中随着HIF-1α蛋白表达的增多,Snail和Vimentin蛋白的表达逐渐增多,E-cadherin蛋白的表达逐渐减少(P<0.05);150μmol/L CoCl2处理24 h后,侵袭、迁移细胞数目较对照组明显增加。结论 HIF-1α能够通过上调Snail与Vimentin的表达,下调E-cadherin的表达,进而促进FTC EMT的发生。
Objective To investigate the expression of Hypoxia-inducible factor-1α (HIF-1α) and epithelial-mesenchymal transition (EMT) -related proteins in follicular thyroid carcinoma (FTC) Expression and clinical significance. Methods The expressions of HIF-1α, Snail, vimentin and E-cadherin in 42 FTC tissues were detected by immunohistochemical SP method. The effects of different concentrations of CoCl2 FCR WRO cells. The effect of different concentrations of CoCl2 on the mRNA transcription of HIF-1α in WRO cells was detected by RT-PCR. The nuclear HIF-1α protein in WRO cells treated with 150 μmol / L CoCl2 for 24 h was detected by immunofluorescence. Western blot was used to detect the effect of 150μmol / L CoCl2 treatment on the protein expression of HIF-1α, Snail, Vimentin and E-cadherin in WRO cells at different times; Transwell chamber assay was used to detect the invasion and migration ability of WRO cells treated with 150μmol / influences. Results The expressions of HIF-1α, Snail, Vimentin and E-cadherin in 42 FTC tissues were correlated with multiple clinical indicators, and the expressions of 4 were significantly correlated (P <0.05). CoCl2 inhibited the proliferation of WRO cells. With the increase of CoCl2 concentration, the transcription level of HIF-1α mRNA in WRO cells first increased and then stabilized. The HIF-1α protein translocated in WRO cells treated with 150μmol / L CoCl2 for 24 hours. CoCl2-treated WRO cells The expression of Snail and Vimentin protein gradually increased, while the expression of E-cadherin protein decreased gradually with the increase of HIF-1α protein expression (P <0.05). After treated with 150μmol / L CoCl2 for 24 h, Group increased significantly. Conclusion HIF-1α can down-regulate the expression of Snail and Vimentin, down-regulate the expression of E-cadherin and further promote the occurrence of FTC EMT.