中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2011年
11期
941-944
,共4页
杨井金%文芳%曹仁贤%钟警%文格波
楊井金%文芳%曹仁賢%鐘警%文格波
양정금%문방%조인현%종경%문격파
无血清处理%S100A13%成纤维细胞生长因子l%[Ca2+]浓度%钙离子螯合剂
無血清處理%S100A13%成纖維細胞生長因子l%[Ca2+]濃度%鈣離子螯閤劑
무혈청처리%S100A13%성섬유세포생장인자l%[Ca2+]농도%개리자오합제
Serum-deprivation-induced%S100A1 3%Fibroblast growth factor 1%[ Ca2+ ] i%Ca2+-chelating agent
目的 探讨无血清处理人甲状腺髓样癌细胞(TT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Ca2+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Ca2+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TT细胞4h和6h后S100A13及FGF-1蛋白表达降低(P＜0.05或P＜0.0l),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Ca2+荧光显像发现无血清处理TT细胞23 min内[Ca2+]i保持相对稳定,23 min后[Ca2+]i迅速上升达到峰值1.6μmol/L,第40 win后下降至一个较低水平,第40 min至第60 min[ Ca2+]i接近0.3～0.6 μmol/L.1h内TT细胞平均[Ca2+]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[Ca2+]i变化有关；Ca2+螯合剂EGTA、BAPTA-AM能有效拮抗无血清处理引起的TT细胞[Ca2+]i升高,并抑制S100A13及FGF-1蛋白的释放.
目的 探討無血清處理人甲狀腺髓樣癌細胞(TT細胞)內S100A13及成纖維細胞生長因子1( FGF-1)蛋白釋放的機製,初步闡明Ca2+在S100A13及FGF-1蛋白釋放過程中的作用.方法 Western印跡檢測無血清處理TT細胞S100A13及FGF-1蛋白錶達；ELISA檢測培養上清液FGF-1蛋白濃度；激光共聚焦顯微鏡實時動態檢測無血清處理TT細胞1h內Ca2+濃度([Ca2+]i)變化；間接免疫熒光觀察TT細胞內S100A13及FGF-1蛋白熒光分佈.結果 無血清處理TT細胞4h和6h後S100A13及FGF-1蛋白錶達降低(P＜0.05或P＜0.0l),而TT細胞培養上清液中FGF-1蛋白濃度升高(P＜0.05或P＜0.01).激光共聚焦Ca2+熒光顯像髮現無血清處理TT細胞23 min內[Ca2+]i保持相對穩定,23 min後[Ca2+]i迅速上升達到峰值1.6μmol/L,第40 win後下降至一箇較低水平,第40 min至第60 min[ Ca2+]i接近0.3～0.6 μmol/L.1h內TT細胞平均[Ca2+]i明顯高于正常培養組、EGTA組和BAPTA-AM組.加入鈣離子螯閤劑EGTA組和BAPTA-AM組TT細胞內的S100A13、FGF-1蛋白錶達無明顯下降.結論 無血清處理誘導TT細胞內S100A13及FGF-1蛋白的釋放與[Ca2+]i變化有關；Ca2+螯閤劑EGTA、BAPTA-AM能有效拮抗無血清處理引起的TT細胞[Ca2+]i升高,併抑製S100A13及FGF-1蛋白的釋放.
목적 탐토무혈청처리인갑상선수양암세포(TT세포)내S100A13급성섬유세포생장인자1( FGF-1)단백석방적궤제,초보천명Ca2+재S100A13급FGF-1단백석방과정중적작용.방법 Western인적검측무혈청처리TT세포S100A13급FGF-1단백표체；ELISA검측배양상청액FGF-1단백농도；격광공취초현미경실시동태검측무혈청처리TT세포1h내Ca2+농도([Ca2+]i)변화；간접면역형광관찰TT세포내S100A13급FGF-1단백형광분포.결과 무혈청처리TT세포4h화6h후S100A13급FGF-1단백표체강저(P＜0.05혹P＜0.0l),이TT세포배양상청액중FGF-1단백농도승고(P＜0.05혹P＜0.01).격광공취초Ca2+형광현상발현무혈청처리TT세포23 min내[Ca2+]i보지상대은정,23 min후[Ca2+]i신속상승체도봉치1.6μmol/L,제40 win후하강지일개교저수평,제40 min지제60 min[ Ca2+]i접근0.3～0.6 μmol/L.1h내TT세포평균[Ca2+]i명현고우정상배양조、EGTA조화BAPTA-AM조.가입개리자오합제EGTA조화BAPTA-AM조TT세포내적S100A13、FGF-1단백표체무명현하강.결론 무혈청처리유도TT세포내S100A13급FGF-1단백적석방여[Ca2+]i변화유관；Ca2+오합제EGTA、BAPTA-AM능유효길항무혈청처리인기적TT세포[Ca2+]i승고,병억제S100A13급FGF-1단백적석방.
Objective To investigate the effect of serum-deprivation on the changes of [ Ca2+ ] i and the protein release of S100A13 and fibroblast growth factor 1 ( FGF-1 ) in thyroid cancer TT cell,and to reveal the role of Ca2+in the protein release of S100Al3 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 [ Ca2+ ] i in TT cells under serum-deprivation were detected by confocal laser scanning microscopy.Then,the effect of EGTA( 2.5 mmol/ L),BAPTA-AM (2.5 μmol/L)on distributions of the fluorescence of S100AI 3 and FGF-1 in TT cells under serumdeprivation for6 h were detected by indirect immunofluorescence.Results The expressions of FGF-1 and S100A13 in TT cells after serum-deprivation for4 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 Ca2+ imaging indicated that [ Ca2+ ] i of serum-deprivation TT cells maintained the relative stabilization within 23 win,but the rapid rise of [ Ca2+ ] i achieved peak value 1.6 μmol/L after 30 min,and remained stable for about 17 win,and thereafter 40 win slowly dropped to a low level From 40 win to 60 win the [ Ca2+ ] i was about 0.3-0.6 μ mol/L.The average [ Ca2+ ] 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 [ Ca2+ ]i.Both Ca2+-chelating agents EGTA and BA PTA-AM are able to inhibit the rise of [ Ca2+ ] i and release of S100A 13 and FGF-1 from TT cells under serum-deprivation.
