囊性纤维化跨膜传导调节因子在人破骨细胞中表达及其对破骨细胞凋亡的影响

Expression of cystic fibrosis transmembrane conduction regulator in human osteoclasts and its effect on apoptosis of osteoclasts

目的探讨囊性纤维化跨膜传导调节因子(Cystic fibrosis transmembrane conductance regulator,CFTR)在破骨细胞中的表达及其对破骨细胞凋亡的影响。方法利用Real time PCR和Western Blot检测人破骨细胞中CFTR表达,运用基因工程手段构建含有GFP的CFTR超表达载体,应用流式技术分析CFTR超表达后对破骨细胞凋亡的影响。利用Real time PCR和Western Blot检测破骨细胞中Fas/FasL的表达。结果 Real time PCR和Western Blot结果显示,体外培养的人破骨细胞中有CFTR的表达。CFTR高表达后,破骨细胞的凋亡率明显升高,凋亡相关蛋白Fas/FasL表达也明显升高。结论 CFTR可通过促进Fas/FasL的高表达,进而促进破骨细胞凋亡。

Abstract:Objective To study the expression of cystic fibrosis transmembrane conduction regulator (CFTR) in osteoclasts and its effect on the apoptosis of osteoclasts. Methods The expression of CFTR in human osteoclasts was detected by real time PCR and Western blot. The CFTR overexpression vector containing GFP was constructed by means of genetic engineering, and flow cytometry was used to analyze the effect of overexpression of CFTR on apoptosis of osteoclasts. Real time PCR and Western blot were used to detect the expression of Fas/FasL in osteoclasts. Results Real time PCR and Western blot showed CFTR was expressed in cultured human osteoclasts. In addition, overexpression of CFTR increased the apoptosis rate of osteoclasts, and also upregulated the expression of apoptosis-related protein Fas/FasL. Conclusion CFTR promotes apoptosis of osteoclasts by promoting the expression of Fas/FasL.