Li Yuan, Wang Yanyan, Song Zhaopu, Lu Kai, Chen Wenwen, Ma Yuanyuan, Ding Hui, Li Xiaofang, Li Xiuling, Sun Suofeng
Department of Traditional Chinese Medicine, The Third Affiliated Hospital Affiliated of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450003, China.
Department of Gastroenterology, Zhengzhou University People's Hospital, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, China.
J Oncol. 2023 Jan 23;2023:3624635. doi: 10.1155/2023/3624635. eCollection 2023.
The occurrence of liver fibrosis and fibrosis-related liver cancer is the reason for the increase in morbidity and mortality worldwide. Transforming growth factor-2 (TGF-2) is an important mediator of chronic liver fibrosis. This study aims to find the molecular mechanism that mediates HBV infection and induces TGF-2 and verifies that CREB binding protein acetylation mediates HBV infection and induces TGF-2 expression.
The acetylated proteins were extracted from HepG2-NTCP cells and HBV-infectedHepG2-NTCP cells. The acetylated proteins were screened by modification enrichment technology and database search. Protein annotation, motif analysis of modification sites, and protein function enrichment analysis of these proteins were performed to roughly clarify the location and function of these acetylated modification proteins in cells. Acylated proteins enriched in the TGF- pathway were obtained by KEGG pathway enrichment analysis. The effect of the selected acetylated modification protein on the TGF- pathway was verified by experiments, that is, the target protein gene was knocked out by siRNA, and the expression level of the TGF-2 was detected by qRT-PCR.
Proteins were extracted from HepG2-NTCP cells and HepG2-NTCP cells infected with HBV, and differential acetylation modification proteins were screened. The target protein CREB binding protein was screened by modification enrichment technology and database search. The aggregation analysis of TGF- pathway showed that CREB binding protein was acetylated at amino acid positions 434 and 439, and enriched in the TGF- signaling pathway. siRNA targeting CREB binding protein was transfected, and the expression of TGF-2 in cells was detected by qRT-PCR and western blot, respectively. It was verified that HBV infection-inducedCREB-binding protein acetylation regulated the high expression of TGF-2.
After HBV infection, CREBBP acetylation was up-regulated, which promoted the high expression of TGF-2.
肝纤维化及纤维化相关肝癌的发生是全球发病率和死亡率上升的原因。转化生长因子-2（TGF-2）是慢性肝纤维化的重要介质。本研究旨在寻找介导HBV感染并诱导TGF-2的分子机制，并验证CREB结合蛋白乙酰化介导HBV感染并诱导TGF-2表达。
从HepG2-NTCP细胞和HBV感染的HepG2-NTCP细胞中提取乙酰化蛋白。通过修饰富集技术和数据库搜索筛选乙酰化蛋白。对这些蛋白进行蛋白质注释、修饰位点的基序分析以及蛋白质功能富集分析，以大致阐明这些乙酰化修饰蛋白在细胞中的定位和功能。通过KEGG通路富集分析获得富集于TGF-通路的酰化蛋白。通过实验验证所选乙酰化修饰蛋白对TGF-通路的影响，即通过siRNA敲除靶蛋白基因，并通过qRT-PCR检测TGF-2的表达水平。
从HepG2-NTCP细胞和感染HBV的HepG2-NTCP细胞中提取蛋白质，筛选出差异乙酰化修饰蛋白。通过修饰富集技术和数据库搜索筛选出靶蛋白CREB结合蛋白。TGF-通路的聚集分析表明，CREB结合蛋白在氨基酸位置434和439处发生乙酰化，并富集于TGF-信号通路。转染靶向CREB结合蛋白的siRNA，分别通过qRT-PCR和western blot检测细胞中TGF-2的表达。验证了HBV感染诱导的CREB结合蛋白乙酰化调节TGF-2的高表达。
HBV感染后，CREBBP乙酰化上调，促进了TGF-2的高表达。
