中文摘要：

目的探讨无血清处理人甲状腺髓样癌细胞（TY细胞）内S100A13及成纤维细胞生长因子1（FGF—1）蛋白释放的机制，初步阐明Ca^2＋在S100A13及FGF-1蛋白释放过程中的作用。方法Western印迹检测无血清处理TT细胞S100A13及FGF-1蛋白表达；ELISA检测培养上清液FGF—1蛋白浓度；激光共聚焦显微镜实时动态检测无血清处理TT细胞1h内Ca^2＋浓度（[Ca2^＋]i）变化；间接免疫荧光观察TT细胞内S100A13及FGF-1蛋白荧光分布。结果无血清处理111细胞4h和6h后S100A13及FGF-1蛋白表达降低（P〈0．05或P〈0．01），而TT细胞培养上清液中FGF-1蛋白浓度升高（P〈0．05或P〈0．01）。激光共聚焦Ca^2＋荧光显像发现无血清处理TT细胞23min内[Ca^2＋]i保持相对稳定，23min后[Ca^2＋]i迅速上升达到峰值1．6μmol／L，第40min后下降至一个较低水平，第40min至第60min[Ca^2＋]i接近0．3—0．6μmol／L。1h内TT细胞平均[Ca^2＋]i明显高于正常培养组、EGTA组和BAPTA—AM组。加入钙离子螯合剂EGTA组和BAPTA—AM组TT细胞内的S100A13、FGF-1蛋白表达无明显下降。结论无血清处理诱导，TT细胞内S100A13及FGF-1蛋白的释放与[Ca^2＋]i变化有关；Ca^2＋螯合剂EGTA、BAPTA—AM能有效拮抗无血清处理引起的TT细胞[CA^2＋]i升高，并抑制S100A13及FGF-1蛋白的释放。

英文摘要：

Objective To investigate the effect of serum-deprivation on the changes of [ Ca^2＋ ] i and the protein release of S100A13 and fibroblast growth factor 1 （ FGF-1 ） in thyroid cancer Tr cell, and to reveal the role of Ca^2＋in the protein release of S100A13 and FGF-1. Methods The protein expressions of FGF-1 and S100A13 in TT cells under serum-deprivation were detected by Western blot. The released FGF-1 protein from TT cells in the supernatant fluid was detected by ELISA. Realtime dynamic examinations on the change of 1 h [ Ca^2＋ ] i in Tr cells under serum-deprivation were detected by confoeal laser scanning microscopy. Then, the effect of EGTA（ 2. 5 mmol/ L） ,BAPTA-AM（2. 5 μmol/L）on distributions of the fluorescence of S100A13 and FGF-1 in TT cells under serumdeprivation for 6 h were detected by indirect immunofluorescence. Results The expressions of FGF-1 and S100A13 in TT cells after serum-deprivation for 4 h and 6 h were reduced（ P〈0.05 or P〈0.01 ）, but the released FGF-1 protein from TT cells in the supernatant fluid was elevated （ P 〈 0. 05 or P 〈 0.01 ）. Confocal laser scanning of Ca^2＋ imaging indicated that [ Ca^2＋ ] i of serum-deprivation TT cells maintained the relative stabilization within 23 min, but the rapid rise of [ Ca^2＋ ] i achieved peak value 1.6 μmol/L after 30 min, and remained stable for about 17 min, and thereafter 40 min slowly dropped to a low level. From 40 rain to 60 min the [ Ca^2＋ ] i was about 0. 3-0. 6 μmol/L The average [ Ca^2＋ ] i was higher than that in normal group, EGTA group, and BAPTA-AM group within 1 h. The protein expressions of S100A13 and FGF-1 did not drop obviously in EGTA group and BAPTA-AM group. Conlusion The release of S100A13 and FGF-1 from TT cell under serum-deprivation is possibly related with the change of [ Ca^2＋ ] i. Both Ca^2＋-chelating agents EGTA and BAPTA-AM are able to inhibit the rise of [ Ca^2＋ ] i and release of S100A13 and FGF-1 from Tr cells under serum-deprivation.