摘要
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 and clinical significance of hypoxia-inducible factor-1α（HIF-1α）and epithelial-mesenchymal transition（EMT）-related protein in human follicular thyroid carcinoma（FTC）.Methods The expressions of HIF-1α,Snail,Vimentin and E-cadherin in 42 FTC specimens were determined by immunohistochemistry assay with SP staining.The effect of cobalt chloride at various concentrations on proliferation activity of FTC WRO cells was determined by MTT method,while that on transcription of HIF-1α mRNA in WRO cells by RT-PCR.The nucleus location of HIF-1α in WRO cells 24 h after treatment with 150 μmol / L cobalt chloride was observed by IFA.The expression levels of HIF-1α,Snail,Vimentin and E-cadherin in WRO cells after treatment with 150 μmol / L cobalt chloride for various hours were determined by Western blot,while the invasion and metastasis of the cells 24 h after treatment by Transwell assay.Results In the 42 specimens,the expressions of HIF-1α,Snail,Vimentin and E-cadherin were correlated with several clinical indicators,between which significant correlations were observed（P 0.05）.The proliferation activity of WRO cells was suppressed by cobalt chloride,and the transcription level of HIF-1α increased with the increasing concentration of cobalt chloride at first then was stable.Translocation of HIF-1α into nucleus was observed in WRO cells 24 h after treatment with 150 μmol / L cobalt chloride.The expressions of Snail and Vimentin in cobalt chloride-treated WRO cells increased gradually,while that of E-cadherin decreased gradually,with the increasing expression of HIF-1α（P 0.05）.The number of invasive and metastatic WRO cells 24 h after treatment with 150 μmol / L cobalt chloride increased significantly as compared with that of control cells.Conclusion HIF-1α promoted the EMT of FTC by up-regulating the expressions of Snail and Vimentin and down-regulating that of E-cadherin.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第12期1671-1675,1679,共6页
Chinese Journal of Biologicals
基金
国家自然科学基金面上项目(81072183)
重庆市自然科学基金重点项目(CSTC
2011BA5038)
重庆医科大学重点科研课题(XBZD201002)
关键词
低氧诱导因子-1Α
甲状腺滤泡状癌
上皮间质转化
Hypoxia-inducible factor-1α（HIF-1α）
Follicular thyroid carcinoma（FTC）
Epithelial-mesenchymal transition（EMT）
