摘要
目的探讨抑制干扰素诱导蛋白16(IFI16)表达后对人脑血管外膜成纤维细胞(HBVAF)增殖、凋亡与迁移的影响及其可能机制。方法培养HBVAF分3组,未经处理的HBVAF作为空白对照组,转染非特异性小干扰RNA(siRNA)的HBVAF作为阴性对照组,转染IFI16siRNA的HBVAF作为IFI16siRNA组。应用蛋白免疫印迹和实时荧光定量聚合酶链反应(PCR)测定细胞中IFI16、p53、p21蛋白和mRNA表达水平。用四甲基偶氮唑盐(MTT)比色法检测细胞活力,流式细胞术测定细胞凋亡,Transwell法测定细胞迁移情况。结果与阴性对照组比较,转染IFI16siRNA后,HBVAF中IFI16、p53及p21蛋白和mRNA表达水平下调,IFI16siRNA组MTT吸光度值增高(0.70±0.01比0.65±0.01,P<0.05),IFI16siRNA组、阴性对照组与空白对照组之间在细胞迁移数目与凋亡比例上差异无统计学意义。结论抑制IFI16表达可促进HBVAF增殖,其机制可能部分与抑制p53及p21表达有关。
Objective To observe the effects tion, apoptosis and migration of human brain were cultured and divided into three groups of inhibiting interferon-inducible protein 16 （IFI16） on the prolifera- vascular adventitial fibroblasts（HBVAF）. Methods The HBVAF Untreated HBVAF served as the blank control group（ blank control）, HBVAF transfected nonspecific small interference RNA（siRNA） served as negative control group, and which trans- fected IFI16 siRNA as IFI16 siRNA group. The protein and mRNA levels of IFI16, p53, p21 were measured by Western blot and real-time polymerase chain reaction（PCR）. Methylthiazolyldiphenyl-tetrazolium hromide（MTT） was used to detect the cell proliferation of the HBVAF. Cell apoptosis was analyzed by flow cytometry, while the migration was analyzed by transwell chamber. Results Compared with negative control group, the expression lev- els of IFI16, p53, p21 protein and mRNA were decreased in the HBVAF after transfected IFI16 siRNA, which in- creasing the absorbance value of the MTT （ 0.70±0.01 vs 0.65±0.01, P（ 0.05 ）, but there was no significant difference on cell migration and apoptosis between negative control group and IFI16 siRNA group. Conclusion Inhibiting expression of IFI16 can promote HBVAF cell growth, which may he related to the inhibition of p53 and p21 expression.
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2017年第4期358-362,共5页
Chinese Journal of Hypertension
基金
国家自然科学基金项目(81260030)
关键词
干扰素诱导蛋白
血管外膜成纤维细胞
增殖
interferon-inducible protein
vascular adventitial fibroblasts
proliferation
