p65通过调控脂代谢影响HepG2细胞增殖研究*

doi:10.3969/j.issn.1672-5069.2025.02.004

实用肝脏病杂志 ›› 2025, Vol. 28 ›› Issue (2): 173-177.doi: 10.3969/j.issn.1672-5069.2025.02.004

p65通过调控脂代谢影响HepG2细胞增殖研究^*

李全维, 高明慧, 寇卜心, 柴梦音, 霍云飞, 豆双双, 庞丽君, 石英, 刘晓霓

100069 北京市首都医科大学附属北京佑安医院北京肝病研究所

收稿日期:2024-08-02 出版日期:2025-03-10 发布日期:2025-03-11
通讯作者: 刘晓霓,E-mail:liuxiaoni888@ccmu.edu.cn
作者简介:李全维,男,28岁,硕士研究生。E-mail:quanweili@mail.ccmu.edu.cn; 共同第一作者:高明慧,女,24岁,硕士研究生。E-mail:minghui512484@163.com
基金资助:
^*国家自然科学基金资助项目(编号:82003200);北京市属医学科研院所公益发展改革试点项目(编号:京医研2021-10);北京市自然科学基金资助项目(编号:7192084);首都卫生发展科研专项基金资助项目(编号:2020-2-1152)

p65 affects proliferation of HepG2 cells in in vitro by regulating lipid metabolism

Li Quanwei, Gao Minghui, Kou Buxin, et al

Beijing Institute of Hepatology, You'an Hospital, Capital Medical University, Beijing 100069, China

Received:2024-08-02 Online:2025-03-10 Published:2025-03-11

摘要/Abstract

摘要： 目的 p65作为NF-κB家族重要的转录因子,在肝细胞癌进展过程中发挥着重要作用。本研究旨在探索p65对HepG2细胞脂代谢调控的影响。方法应用数据库分析p65表达与HCC患者预后的关系。采用ChIP-seq和RNA-seq技术,采用生物信息学分析探究HepG2细胞p65 DNA结合谱,使用流式细胞术检测p65对HepG2细胞增殖的影响。采用实时定量PCR法(qRT-PCR)和免疫印记(Western blot,WB)法验证p65对脂代谢途径关键基因及其蛋白表达的影响,采用流式细胞术和共聚焦显微镜检测p65对HepG2细胞脂代谢的影响。结果对UCSC Xena和GEPIA数据库分析显示,HCC癌组织p65常呈高表达,并与不良预后相关;与对照组比,敲低p65抑制HepG2细胞增殖,而过表达p65则促进HepG2细胞增殖;对ChIP-seq和RNA-seq数据综合分析共得到了205个共同基因,且富集在代谢途径中的基因最多,其中脂代谢相关关键基因包括ACSM2A、ACSM2B、ACSM3、ACSM5和HMGCS2等;p65敲低后ACSM5和HMGCS2 mRNA及其蛋白表达均显著降低,而p65过表达后ACSM5和HMGCS2 mRNA及其蛋白表达均显著升高;p65敲低能促进HepG2细胞脂质积累,而p65过表达则抑制HepG2细胞脂质的积累。结论 p65通过上调ACSM5和HMGCS2表达调控脂质代谢,促进HepG2细胞增殖。这一发现为揭示p65调控肝细胞癌细胞脂质代谢的机制提供了研究线索。

关键词: 肝细胞癌, p65, ChIP-seq, RNA-seq, 细胞增殖, 脂代谢, 体外

Abstract: Objective As an important transcription factor of NF-κB family, p65 plays a pivotal roles in progression of hepatocellular carcinoma (HCC). This study aimed to explore effect of p65 on regulation of lipid metabolism in HepG2 cells in vitro. Methods In this study, relationship between p65 and prognosis of patients with HCC was investigated in UCSC Xena and GEPIA database. ChIP-seq and RNA-seq technologies were conducted to explore DNA binding profile of p65 in HepG2 cells through bioinformatics analysis, and flow cytometry was applied to detect effect of p65 on the proliferation of HepG2 cells. p65 on expression of key genes and their proteins were detected by real-time quantitative PCR (qRT-PCR) and Western blot (WB), and effect of p65 on lipid metabolism in HepG2 cells was determined by flow cytometry and confocal microscopy fluorescence. Results Data analysis from database showed that p65 was often highly expressed in patients with HCC and the intensified expression was associated with poor prognosis of patients with HCC; p65 knockdown inhibited the proliferation of HepG2 cells, and overexpression of p65 boasted the proliferation of HepG2 cells as compared to in control; by comprehensive analysis of ChIP-seq and RNA-seq data, 205 common genes were obtained, and the most abundant genes were in the metabolic pathway, among which the key genes including ACSM2A, ACSM2B, ACSM3, ACSM5 and HMGCS2, were found to be related to lipid metabolism; ACSM5 and HMGCS2 mRNA and their protein were significantly decreased after p65 was knocked down, while they significantly increased after p65 was overexpressed; p65 knockdown promoted lipid accumulation, while p65 overexpression inhibited lipid accumulation in HepG2 cells. Conclusion p65 regulates lipid metabolism by up-regulating the expression of ACSM5 and HMGCS2 and promotes the proliferation of HepG2 cells, which provides research clues for the mechanism of p65 regulation of lipid metabolism in hepatocellular carcinoma.

Key words: Hepatocellular carcinoma, p65, ChIP-seq, RNA-seq, Cell proliferation, Lipid metabolism, In vitro

李全维, 高明慧, 寇卜心, 柴梦音, 霍云飞, 豆双双, 庞丽君, 石英, 刘晓霓. p65通过调控脂代谢影响HepG2细胞增殖研究^*[J]. 实用肝脏病杂志, 2025, 28(2): 173-177.

Li Quanwei, Gao Minghui, Kou Buxin, et al. p65 affects proliferation of HepG2 cells in in vitro by regulating lipid metabolism[J]. Journal of Practical Hepatology, 2025, 28(2): 173-177.

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p65通过调控脂代谢影响HepG2细胞增殖研究^*

p65 affects proliferation of HepG2 cells in in vitro by regulating lipid metabolism

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