[1]王俊杰,姜艳,Soulixay Senouthai,等.血管紧张素Ⅱ通过上调富含半胱氨酸蛋白61 表达诱导HEK293T细胞凋亡[J].南方医科大学学报,2019,(07):810.[doi:10.12122/j.issn.1673-4254.2019.07.10]
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血管紧张素Ⅱ通过上调富含半胱氨酸蛋白61 表达诱导HEK293T细胞凋亡()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2019年07期
页码:
810
栏目:
出版日期:
2019-07-15

文章信息/Info

Title:
Angiotensin II induces apoptosis of HEK293T cells by up-regulating Cyr61 expression
作者:
王俊杰姜艳Soulixay Senouthai付冬冬尤燕舞
关键词:
HEK293T细胞CRISPR/Cas9Cyr61AngⅡ凋亡
Keywords:
HEK293T cells CRISPR/Cas9 Cyr61 angiotensin II apoptosis
DOI:
10.12122/j.issn.1673-4254.2019.07.10
摘要:
目的探讨富含半胱氨酸蛋白61(Cyr61)在血管紧张素Ⅱ(AngⅡ)诱导HEK293 细胞功能中的改变及作用。方法 HEK293T细胞于体外培养,并应用CRISPR/Cas9技术敲低HEK293T细胞Cyr61基因。将细胞分为四组:(1)对照组;(2)Cyr61 敲低组;(3)AngⅡ组;(4)AngⅡ+Cyr61敲低组,以10-7 mol/L AngⅡ处理细胞,48 h收集细胞用流式细胞仪检测细胞凋亡,通过 实时荧光定量PCR(qRT-PCR)技术及蛋白质免疫印迹技术检测细胞Cyr61及Bcl-2的mRNA及蛋白表达量。结果Cyr61敲低 组Cyr61蛋白水平较正常对照组明显下降(P<0.05),AngⅡ干预48 h 后Cyr61蛋白和mRNA表达水平显著升高(P<0.05),Bcl-2 蛋白和mRNA表达水平明显下降(P<0.05),敲低Cyr61 后Bcl-2 蛋白和mRNA表达水平明显升高(P<0.05),与对照组凋亡率 (11.88±1.46)%相比,Cyr61敲低组凋亡率为(3.87±0. 83)%,AngⅡ干预组HEK293T细胞凋亡率为(26.94±3.73)%(P<0.05),与 AngⅡ干预组相比,Cyr61 敲低+AngⅡ组凋亡率为(15.76±1.31)%(P<0.05)。结论Cyr61 表达量的上调与AngⅡ诱导的 HEK293T细胞损伤有关,下调Cyr61的表达可以有效地保护AngⅡ诱导的细胞损伤。
Abstract:
Objective To investigate the role of Cyr61 in angiotensin II (AngII)-induced functional changes in HEK293 cells and explore the mechanism. Methods Cyr61 knockdown in cultured HEK293T cells was achieved by transfection of the cells with CRISPR/Cas9 KO plasmid. The changes in apoptosis and expression levels of Cyr61 and Bcl-2 in the cells with or without Cyr61 knockdown in response to treatment with 10-7 mol/L AngII for 48 h were analyzed using flow cytometry, qRT-PCR and Western blotting. Results The cells with Cyr61 knockdown showed significantly decreased expression of Cyr61 protein as compared with the control cells (P<0.05). AngII treatment for 48 h significantly increased the expression of Cyr61 and lowers the expression of Bcl-2 at both the protein and mRNA levels in HEK293T cells. In HEK293T cells with Cyr61 knockdown, AngII treatment resulted in significantly increased expression of Bcl-2 in HEK293T cells as compared with that of the control group (P<0.05). AngII treatment caused significantly increased apoptotic rate in HEK293T cells as compared with the cells with Cyr61 knockdown [(26.94 ± 3.73)% vs (3.87 ± 0.83)% , P<0.05), and the apoptosis rate was significantly lowered to (15.76 ± 1.31)% in HEK293T cells with Cyr61 knockdown following AngII treatment (P<0.05). Conclusion The up-regulation of Cyr61 expression is related with AngII-induced injury in HEK293T cells, and down-regulating Cyr61 expression can effectively protect HEK293T cells against AngII-induced injury.

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[2]周燕霞,胡韦维,张虹洋,等.利用CRISPR/Cas9 技术构建敲除G6PD基因c.392G>T(p.131G>V)突变位点的HEK293T稳定细胞株[J].南方医科大学学报,2019,(03):320.[doi:10.12122/j.issn.1673-4254.2019.03.10]

更新日期/Last Update: 1900-01-01