摘要
目的研究低氧诱导金属硫蛋白(metallothioneins,MTs)对甲状腺乳头状癌(papillary thyroid carcinoma,PTC)上皮间质转化(epithelial-mesenchymal transition,EMT)的影响。方法以人甲状腺乳头状癌NPA细胞为研究模型,将其分为正常氧组(21%O2)、低氧组(5%O2)和MTs-siRNA干扰低氧组,Western blot、RT-PCR方法检测各组细胞MTs、上皮细胞钙粘蛋白(E-cadherin)和间质细胞波形蛋白(Vimentin)的表达,Transwell/Matrigel方法检测各组细胞侵袭转移细胞数。结果低氧可诱导NPA细胞表达MTs,并伴随有EMT标志性分子E-cadherin表达的下调和Vimentin表达的升高,侵袭细胞数由正常氧组15.6±2.8增加到40.3±5.1,迁移细胞数由25.1±5.6增加到48.7±7.6,均有统计学差异(P<0.01);MTs靶向干扰可有效抑制低氧诱导的MTs蛋白表达,同时E-cadherin表达回升,Vimentin表达下降,侵袭细胞数仅增加到25.7±4.2,迁移细胞数仅增加到35.5±5.4,与低氧组比较均有统计学差异(P<0.05)。结论低氧所诱导表达的MTs对人PTC细胞发生EMT有促进作用,MTs靶向干扰可使EMT受到部分抑制。
Objective To determine the effects of metallothioneins（MTs） induced by hypoxia on epithelial-mesenchymal transition（EMT） in human papillary thyroid carcinoma NPA cells.Methods NPA cells were divided into 3 groups,normoxia（21% O2）,hypoxia（5% O2） and hypoxia with MTs-siRNA.The expression levels of MTs,E-cadherin and Vimentin in the 3 groups of NPA cells were analyzed by Western blotting and RT-PCR.The invasion and metastasis potential of these groups were detected by Transwell/Matrigel assay.Results Hypoxia treatment led to inducible expression of MTs,down-regulated of E-cadherin and up-regulated Vimentin at the same time,which were marker molecules of EMT in NPA cells.Meanwhile,invasive cell population of hypoxia group was increased to 40.3±5.1 from 15.6±2.8 of normoxia group（P0.01）.Migrating cell population was increased to 48.7±7.6 from 25.1±5.6 of normoxia group（P0.01）.However,the inducible expression of MTs was effectively inhibited by transfection of MTs-siRNA vector in NPA cells,followed by increased expression of E-cadherin and decreased expression of Vimentin.Moreover,invasive and migrating cell population of the hypoxia group with MTs-siRNA were only increased to 25.7±4.2 and 35.5±5.4（P0.05）.Conclusion The expression of MTs induced by hypoxia can promote EMT in human papillary thyroid carcinoma.siRNA targeting MTs can partially inhibit EMT promoted by hypoxia.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第7期671-674,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(81072183)
重庆医科大学重点科研项目(XBZD201002)~~
关键词
金属硫蛋白
上皮间质转化
低氧
甲状腺乳头状癌
metallothioneins
epithelial-mesenchymal transition
hypoxia
papillary thyroid carcinoma
