Page 51 - 南京医科大学学报自然科学版

第43卷第8期 南京医科大学学报（自然科学版）
2023年8月 Journal of Nanjing Medical University（Natural Sciences） ·1085 ·

·基础研究·

转录因子AP2A对肾病相关基因Gas6的调控研究

颜 凤，周国平 *
南京医科大学第一附属医院儿科，江苏 南京 210009

［摘 要］ 目的：探究肾脏病相关生长抑制特异性基因6（growth arrest specific 6，Gas6）与肾脏疾病、凋亡的相关关系。研究转
录因子AP⁃2a（transcription factor AP⁃2 alpha，TFAP2A）对基因Gas6的影响及其机制。方法：构建阿霉素肾病细胞模型，检测该
模型中Gas6的表达变化；用Gas6蛋白刺激MPC5细胞，检测肾病相关指标的表达变化。用Gas6蛋白干预HEK⁃293T细胞，检
测细胞凋亡率或凋亡相关指标的表达变化。构建Gas6启动子片段的萤光素酶基因报告重组质粒，检测Gas6基因启动子片段
在 HEK⁃293T 细胞中的活性，预测 Gas6 启动子区的功能性转录因子结合位点并验证，在 HEK⁃293T 细胞中敲低或过表达
TFAP2A后检测Gas6启动子片段活性、mRNA表达、蛋白表达的变化。结果：Gas6在阿霉素肾病细胞模型中高表达，且过量的
Gas6可诱导MPC5细胞中肾病相关指标Nephrin和Podocin与阿霉素肾病细胞模型趋势相同。与对照组相比，Gas6蛋白组的细
胞凋亡明显减少。成功构建有活性的Gas6基因启动子片段萤光素酶基因报告重组质粒，且在此片段内含有TFAP2A的结合位
点。TFAP2A 可在启动子、mRNA 及蛋白水平对 Gas6 正向调控。结论：Gas6 对肾病综合征的发生有促进作用，Gas6 在 HEK⁃
293T细胞中有抗凋亡作用，TFAP2A对Gas6有正向转录调控作用。
［关键词］ Gas6；TFAP2A；启动子；转录调控；肾病综合征；凋亡
［中图分类号］ R692 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）08⁃1085⁃09
doi：10.7655/NYDXBNS20230807

The transcription factor AP2A regulating human nephrosis⁃related gene Gas6

YAN Feng，ZHOU Guoping *
Department of Pediatrics，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210009，China

［Abstract］ Objective：To explore the correlation between nephrosis⁃related growth arrest specific 6（Gas6）and nephrotic syndrome
and apoptosis.The effect of AP⁃2 transcription factor a（TFAP2A）on Gas6 was investigated. Methods：The cell model of adriamycin
nephropathy（AN）was constructed to detect the expression of Gas6，and MPC5 cells were stimulated with Gas6 protein to measure the
expression of nephropathy related indicators.HEK ⁃ 293T cells were intervened with Gas6 protein to measure apoptosis rate or the
expression of apoptosis ⁃ related indicators. The luciferase gene reporter recombinant plasmid of Gas6 promoter was constructed to
measure the activity of Gas6 promoter in HEK⁃293T cells. And the potential transcriptional binding sites were predicted and verified.
The effects of knockdown or overexpression of TFAP2A on Gas6 gene expression were measured at the promoter，mRNA，and protein
levels. Results：Gas6 was highly expressed in AN cell model，and excessive Gas6 may induce the nephropathy⁃related indicators in
MPC5 cells the same trend as that of AN cell model. Apoptosis was significantly reduced in the Gas6 protein group compared to control
in HEK⁃293T cells. The active luciferase reporter plasmid of human Gas6 promoter fragment was successfully constructed，and the
binding sites of TFAP2A were included. TFAP2A positively regulated Gas6 at the promoter，mRNA，and protein levels. Conclusion：
Gas6 promotes the development of nephrotic syndrome. Gas6 is antiapoptotic in HEK⁃293T cells. TFAP2A positively promotes the
expression of human nephrosis⁃related gene Gas6.
［Key words］ Gas6；TFAP2A；promoter；transcriptional regulation；nephrotic syndrome；apoptosis
［J Nanjing Med Univ，2023，43（08）：1085⁃1093］

肾病综合征（nephrotic syndrome，NS）是常见的

［基金项目］ 国家自然科学基金（82270068） 小儿肾脏疾病，指蛋白尿过多，伴有低白蛋白血症、
［1］
∗ 水肿和高脂血症 。儿童NS的病程反复，可并发感
通信作者（Corresponding author），E⁃mail：guopzhou@126.com