摘要
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。
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＆amp;nbsp;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 .
出处
《中华关节外科杂志（电子版）》
CAS
2014年第2期51-53,共3页
Chinese Journal of Joint Surgery(Electronic Edition)
基金
广东省自然科学基金(S2012010009184)
