中华关节外科杂志（电子版）
中華關節外科雜誌（電子版）
중화관절외과잡지（전자판）
CHINESE JOURNAL OF JOINT SURGERY(ELECTRONIC VERSION)
2014年
2期
201-205
,共5页
郑少伟%陈国奋%王健%李郅涵%Shaikh Atik Badshah%李敏%张洋%史占军
鄭少偉%陳國奮%王健%李郅涵%Shaikh Atik Badshah%李敏%張洋%史佔軍
정소위%진국강%왕건%리질함%Shaikh Atik Badshah%리민%장양%사점군
骨肉瘤%增殖%凋亡%钙库依赖性Ca2+内流
骨肉瘤%增殖%凋亡%鈣庫依賴性Ca2+內流
골육류%증식%조망%개고의뢰성Ca2+내류
Osteosarcoma%Proliferation%Apoptosis%Store operated Ca 2+entry
目的：探讨钙库操纵的钙内流（ SOCE）组成蛋白STIM1和Orai1蛋白在骨肉瘤细胞系HOS增殖和凋亡中的作用。方法用shRNA干扰技术抑制STIM1和Orai1蛋白的表达。 Western blot测定STIM1和Orai1的蛋白表达；激光共聚焦检测细胞Ca2＋内流变化；噻唑蓝（ MTT）比色法检测HOS细胞的增殖能力；流式细胞技术检测HOS 细胞的凋亡变化。结果与空白对照组和转染negative-shRNA的对照组相比，转染48 h后STIM1-shRNA 组STIM1蛋白的表达均明显降低（ P＜0.01），Orai1-shRNA组Orai1蛋白的表达也明显降低。抑制STIM1和Orai1蛋白后，细胞Ca2＋内流减少。转染24、48和72 h后，STIM1-shRNA组及Orai1-shRNA组HOS细胞的增殖能力明显下降（ P＜0.01）。流式细胞检测显示，与control组和negative-shRNA组相比，转染48 h后细胞的周期受到明显抑制，而凋亡水平明显升高（P＜0.01）。结论 STIM1和Orai1在骨肉瘤细胞HOS增殖中有重要作用，抑制STIM1和Orai1蛋白表达可以抑制HOS细胞增殖、促进凋亡。
目的：探討鈣庫操縱的鈣內流（ SOCE）組成蛋白STIM1和Orai1蛋白在骨肉瘤細胞繫HOS增殖和凋亡中的作用。方法用shRNA榦擾技術抑製STIM1和Orai1蛋白的錶達。 Western blot測定STIM1和Orai1的蛋白錶達；激光共聚焦檢測細胞Ca2＋內流變化；噻唑藍（ MTT）比色法檢測HOS細胞的增殖能力；流式細胞技術檢測HOS 細胞的凋亡變化。結果與空白對照組和轉染negative-shRNA的對照組相比，轉染48 h後STIM1-shRNA 組STIM1蛋白的錶達均明顯降低（ P＜0.01），Orai1-shRNA組Orai1蛋白的錶達也明顯降低。抑製STIM1和Orai1蛋白後，細胞Ca2＋內流減少。轉染24、48和72 h後，STIM1-shRNA組及Orai1-shRNA組HOS細胞的增殖能力明顯下降（ P＜0.01）。流式細胞檢測顯示，與control組和negative-shRNA組相比，轉染48 h後細胞的週期受到明顯抑製，而凋亡水平明顯升高（P＜0.01）。結論 STIM1和Orai1在骨肉瘤細胞HOS增殖中有重要作用，抑製STIM1和Orai1蛋白錶達可以抑製HOS細胞增殖、促進凋亡。
목적：탐토개고조종적개내류（ SOCE）조성단백STIM1화Orai1단백재골육류세포계HOS증식화조망중적작용。방법용shRNA간우기술억제STIM1화Orai1단백적표체。 Western blot측정STIM1화Orai1적단백표체；격광공취초검측세포Ca2＋내류변화；새서람（ MTT）비색법검측HOS세포적증식능력；류식세포기술검측HOS 세포적조망변화。결과여공백대조조화전염negative-shRNA적대조조상비，전염48 h후STIM1-shRNA 조STIM1단백적표체균명현강저（ P＜0.01），Orai1-shRNA조Orai1단백적표체야명현강저。억제STIM1화Orai1단백후，세포Ca2＋내류감소。전염24、48화72 h후，STIM1-shRNA조급Orai1-shRNA조HOS세포적증식능력명현하강（ P＜0.01）。류식세포검측현시，여control조화negative-shRNA조상비，전염48 h후세포적주기수도명현억제，이조망수평명현승고（P＜0.01）。결론 STIM1화Orai1재골육류세포HOS증식중유중요작용，억제STIM1화Orai1단백표체가이억제HOS세포증식、촉진조망。
Objective To investigate the role of STIM 1 and Orai1 in the proliferation and apoptosis of HOS osteosarcoma cells .Methods The protein expressions of STIM 1 and Orai1 were inhibited using small RNA interference .The protein expressions of STIM 1 and Orai1 were detected by Western blot .The proliferation of HOS cells was detected by MTT assay .Results Comparing with the control group and the transfection-negative shRNA group , 48 hours after ransfection , the STIM1 protein expression of the STIM1-shRNA group was significantly decreased; the Orai1 protein expression of the Orai1-shRNA group was also obviously decreased .24 h, 48h and 72 h after the transfection , the cell proliferation of the STIM1-shRNA group and Orai1-shRNA group was significantly decreased (P<0.05). The flow cytometry showed that , comparing with the control group and the negative-shRNA group, 48 h after transfection , the cell cycles were significantly inhibited , while the apoptosis levels were significantly increased ( P<0.05 ) .Conclusion STIM1 and Orai1 have an important role in the HOS osteosarcoma cell proliferation:inhibition of the STIM1 and Orai1 proteins can inhibit the proliferation and apoptosis of the HOS cells .
