氯化钴对甲状腺乳头状癌NPA细胞上皮间质转化的影响及其机制.doc

氯化钴对甲状腺乳头状癌NPA细胞上皮间质转化的影响及其机制 贾朝莉，龙方懿，刘智敏，杨俊艳，刘小花，董超然，王 妮（400016重庆，重庆医科大学基础医学院生物化学与分子生物学教研室，分子医学与肿瘤研究中心） [摘要] 目的 研究氯化钴（CoCl2）诱导的低氧对甲状腺乳头状癌NPA细胞上皮间质转化的影响，初步探讨HIF-1a在其中的机制。方法 MTT检测CoCl2（50、100、150、200、250μmol/L）对细胞活力的影响，倒置显微镜观察低氧（150 μmol/L的CoCl2分别作用0、12、24、48 h）对细胞形态的影响，Western blot检测低氧（150 μmol/L的CoCl2分别作用0、3、6、12、24 h）对HIF-1a、E-cadherin和Vimentin表达的影响，Transwell法检测低氧（150 μmol/L的CoCl2分别作用0、6 h）对细胞侵袭、转移的影响，用免疫荧光对HIF-1a蛋白进行亚细胞定位。结果 低氧模拟剂CoCl2可抑制NPA细胞活力。随着CoCl2低氧处理时间的延长，NPA细胞逐渐获得间质细胞的形态特征，HIF-1a、Vimentin表达在0～6 h间逐渐增加，随后开始下降，E-cadherin表达持续降低，低氧0 h与6 h相比各蛋白表达均具有统计学差异(P＜0.01)。低氧组侵袭、转移细胞数目较对照组明显增加(P＜0.01)，同时低氧组细胞HIF-1a发生核转位。 结论 CoCl2模拟化学性低氧可促进NPA细胞上皮间质转化及侵袭、转移，其分子机制可能涉及HIF-1a介导的信号通路对E-cadherin、Vimentin基因表达的调控。 [关键词] 低氧诱导因子-1a；上皮间质转化；甲状腺乳头状癌 [中图法分类号] R736.1；R364.4；R730.2 [文献标志码] A Effect of CoCl2 on epithelial-mesenchymal transition of human papillary thyroid carcinoma cells Jia Chaoli，Long Fangyi，Liu Zhimin，Yang Junyan，Liu Xiaohua，Dong Chaoran，Wang Ni　（Department of Biochemistry and Molecular Biology，Institute of Molecular Medicine and Cancer，College of Basic Medical Sciences，Chongqing Medical University，Chongqing，400016，China） [Abstract] Objective To determine the effect and mechanism of HIF-1a on epithelial-mesenchymal transition in human papillary thyroid carcinoma. Methods Cellullar vitality was evaluated by MTT. Morphological changes of NPA cells were observed under inverted microscope. The expression levels of HIF-1a、E-cadherin and Vimentin in each group of NPA cells were analyzed by Western blotting. The invasion and metastasis potential of each group was detected by transwell assay. Immunofluorescence was used to detect whether HIF-1a transfers in the NPA cell nucleus. Results NPA cells vitality was suppressed by CoCl2.NPA cells in low oxygen group gradually acquired the morphological characteristics of mesenchymal cells. The expression levels of HIF-1a were rised with the increase of hypoxia time. The expression of E-cadherin was down-regulated in low oxyge