抑制干扰素诱导蛋白16表达减少人主动脉外膜成纤维细胞凋亡
发布时间：2018-11-10 16:51
【摘要】：目的:研究干扰素诱导蛋白16(IFI16)表达抑制后对人主动脉外膜成纤维细胞(HAAFs)凋亡的影响。方法:应用IFI16基因小干扰RNA(siRNA)转染HAAFs使IFI16基因沉默(IFI16-siRNA组),以非特异性siRNA转染作为其转染阴性对照组(Con-siRNA组),未经处理的HAAFs作为未干预阴性对照组(Neg组)。应用流式细胞仪检测细胞周期及凋亡,实时定量逆转录-聚合酶链反应(Realtime qRT-PCR)检测细胞中IFI16 mRNA表达变化,蛋白免疫印迹(Western blotting)检测IFI16、抑癌因子(p53)、细胞周期依赖性蛋白激酶抑制因子p21~(WAF)(p21)、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)及Bcl-2蛋白表达。结果:IFI16-siRNA组与Con-siRNA组、Neg组相比,细胞凋亡率[(3.33±0.41)%vs(7.42±1.51)%(、6.49±1.10)%,P0.05]与G_0/G_1期细胞比例[(56.64±4.77)%vs(69.67±3.54)%、(68.29±4.14)%,P0.05]减少;而S期细胞比例[(25.23±5.19)%vs(13.76±2.07)%、(14.04±3.00)%,P0.05]增加;伴随着IFI16mRNA表达减少(P0.05),以及IFI16-siRNA组IFI16、p53、p21、Bax蛋白表达量减少(P0.05);同时Bcl-2蛋白表达量增加(P0.05)。结论:抑制IFI16表达可减少HAAFs细胞凋亡,促进细胞周期G_1/S转换,其机制部分与抑制p53/p21信号通路及调控Bax/Bcl-2表达有关。
[Abstract]:Aim: to investigate the effect of inhibition of interferon-induced protein 16 (IFI16) expression on (HAAFs) apoptosis in human aortic adventitial fibroblasts. Methods: IFI16 gene was silenced by IFI16 gene small interfering RNA (siRNA) transfection into HAAFs (IFI16-siRNA group), non-specific siRNA transfection was used as the negative control group (Con-siRNA group), and untreated HAAFs was used as unintervention negative control group (Neg group). Flow cytometry was used to detect cell cycle and apoptosis, real-time quantitative reverse transcription-polymerase chain reaction (Realtime qRT-PCR) was used to detect the expression of IFI16 mRNA, and Western blot (Western blotting) was used to detect IFI16, tumor suppressor factor (p53). Cell cycle dependent protein kinase inhibitor p21 ~ (WAF) (p21), B cell lymphoma-2 (Bcl-2)-related X protein (Bax) and Bcl-2 protein were expressed. Results: compared with Con-siRNA group and Neg group, apoptosis rate of IFI16-siRNA group was (3.33 卤0.41)% vs (7.42 卤1.51)% (6.49 卤1.10)%. The proportion of cells in G_0/G_1 phase [(56.64 卤4.77)% vs, (69.67 卤3.54)%, (68.29 卤4.14)%, P0.05] was decreased. The percentage of S phase cells increased [(25.23 卤5.19)% vs (13.76 卤2.07)%, (14.04 卤3.00)%, P0.05]. With the decrease of IFI16mRNA expression (P0.05) and the decrease of IFI16,p53,p21,Bax protein expression in IFI16-siRNA group (P0.05), the expression of Bcl-2 protein increased (P0.05). Conclusion: inhibiting the expression of IFI16 can reduce the apoptosis of HAAFs cells and promote the cell cycle 1 / S transition. The mechanism is partly related to the inhibition of p53/p21 signaling pathway and the regulation of Bax/Bcl-2 expression.
【作者单位】： 贵州省人民医院贵州医科大学附属人民医院心内科;
【基金】：国家自然科学基金资助项目(81260030) 贵州省科技合作计划项目(黔科合LH字[2015]7159号)
【分类号】：R54
本文编号：2323012
[Abstract]:Aim: to investigate the effect of inhibition of interferon-induced protein 16 (IFI16) expression on (HAAFs) apoptosis in human aortic adventitial fibroblasts. Methods: IFI16 gene was silenced by IFI16 gene small interfering RNA (siRNA) transfection into HAAFs (IFI16-siRNA group), non-specific siRNA transfection was used as the negative control group (Con-siRNA group), and untreated HAAFs was used as unintervention negative control group (Neg group). Flow cytometry was used to detect cell cycle and apoptosis, real-time quantitative reverse transcription-polymerase chain reaction (Realtime qRT-PCR) was used to detect the expression of IFI16 mRNA, and Western blot (Western blotting) was used to detect IFI16, tumor suppressor factor (p53). Cell cycle dependent protein kinase inhibitor p21 ~ (WAF) (p21), B cell lymphoma-2 (Bcl-2)-related X protein (Bax) and Bcl-2 protein were expressed. Results: compared with Con-siRNA group and Neg group, apoptosis rate of IFI16-siRNA group was (3.33 卤0.41)% vs (7.42 卤1.51)% (6.49 卤1.10)%. The proportion of cells in G_0/G_1 phase [(56.64 卤4.77)% vs, (69.67 卤3.54)%, (68.29 卤4.14)%, P0.05] was decreased. The percentage of S phase cells increased [(25.23 卤5.19)% vs (13.76 卤2.07)%, (14.04 卤3.00)%, P0.05]. With the decrease of IFI16mRNA expression (P0.05) and the decrease of IFI16,p53,p21,Bax protein expression in IFI16-siRNA group (P0.05), the expression of Bcl-2 protein increased (P0.05). Conclusion: inhibiting the expression of IFI16 can reduce the apoptosis of HAAFs cells and promote the cell cycle 1 / S transition. The mechanism is partly related to the inhibition of p53/p21 signaling pathway and the regulation of Bax/Bcl-2 expression.
【作者单位】： 贵州省人民医院贵州医科大学附属人民医院心内科;
【基金】：国家自然科学基金资助项目(81260030) 贵州省科技合作计划项目(黔科合LH字[2015]7159号)
【分类号】：R54
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