CAJ | 학술논문

目的论证上皮细胞间质样转化(EMT)能否诱导人甲状腺肿瘤干细胞(CSC)的发生.方法转染缺氧诱导因子(HIF)-1α cDNA诱导甲状腺癌细胞发生EMT,并以Western印迹法、免疫荧光、Transwell及四甲基偶氮唑蓝(MTT)等技术验证EMT分子特征及行为特征的变化,应用荧光激活的流式细胞分选(FACS)技术对比EMT属性变化对细胞株中干细胞分选效率的影响,并鉴定所分选细胞的“干性”分子特征及自我更新和克隆形成能力.结果转染HIF-1α成功诱导人甲状腺癌FTC133细胞完成EMT:E-钙黏蛋白上调,波形蛋白下调,β-连环蛋白核易位,细胞体现出显著增强的体外侵袭和增殖能力.另外,EMT后的FTC133细胞中CSC的分选效率明显升高(0.70％比0.03％,P＜0.05).所分选细胞具有“干性”分子标志,并体现出了典型的自我更新及体外致瘤能力.结论 HIF-lα能够诱导人甲状腺癌细胞完成EMT,进而促使CSC发生.
목적 론증상피세포간질양전화(EMT)능부유도인갑상선종류간세포(CSC)적발생.방법 전염결양유도인자(HIF)-1α cDNA유도갑상선암세포발생EMT,병이Western인적법、면역형광、Transwell급사갑기우담서람(MTT)등기술험증EMT분자특정급행위특정적변화,응용형광격활적류식세포분선(FACS)기술대비EMT속성변화대세포주중간세포분선효솔적영향,병감정소분선세포적“간성”분자특정급자아경신화극륭형성능력.결과 전염HIF-1α성공유도인갑상선암FTC133세포완성EMT:E-개점단백상조,파형단백하조,β-련배단백핵역위,세포체현출현저증강적체외침습화증식능력.령외,EMT후적FTC133세포중CSC적분선효솔명현승고(0.70％비0.03％,P＜0.05).소분선세포구유“간성”분자표지,병체현출료전형적자아경신급체외치류능력.결론 HIF-lα능구유도인갑상선암세포완성EMT,진이촉사CSC발생.
Objective Epithelial-mesenchymal transition (EMT) has been implicated in the initiation and conversion of early stage tumors into invasive malignancies and is associated with the "stemness" of cancer cells.The present study was designed to identify whether EMT induces cancer stem cell generation and tumor progression in human thyroid cancer cells in vitro.Methods FTC133 cells,as EMT-negative cells,were used for EMT induction by hypoxia-inducible factor-1 α (HIF-1α) transfection.And EMT features were then examined by Western blot,immunofluorescent staining,invasion and proliferation assays.Moreover,stem-like side population (SP) cells were sorted with flow cytometry from FTC133 cells before and after EMT.The proportion of SP was compared and stemness,self-renewal and tumorigenecity in vitro were identified in SP cells.Results Overexpression of HIF-1α induced FTC133 cells to undergo EMT.And it down-regulated epithelial marker E-cadherin,up-regulated mesenchymal narker vimentin and caused nucleus translocation of β-catenin and highly invasive and metastatic properties.Most importantly,the induction of EMT promoted proportion of stem-like side population cells (0.70％ vs 0.03％,P ＜ 0.05) with higher sphere formation and clone forming capability in contrast to non-side population cells.Conclusions EMT can induce cancer stem cell generation and tumor progression in thyroid cancers.Further understanding the role of EMT and cancer stem cells in cancer progression may reveal new preventive and therapeutic targets for thyroid cancers.