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

Abstract

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

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.

References

[1] Sun S C. Non-canonical NF-κB signaling pathway. Cell Res, 2011,21(1):71-85.
[2] Sun S C. The non-canonical NF-κB pathway in immunity and inflammation. Nat Rev Immunol, 2017,17(9):545-558.
[3] Shin C, Ito Y, Ichikawa S, et al. MKRN2 is a novel ubiquitin E3 ligase for the p65 subunit of NF-κB and negatively regulates inflammatory responses. Sci Rep, 2017,7:46097.
[4] Zhang T, Ma C, Zhang Z, et al. NF-κB signaling in inflammation and cancer. Med Comm (2020), 2021,2(4):618-653.
[5] Levidou G, Saetta A A, Korkolopoulou P, et al. Clinical significance of nuclear factor (NF)-kappaB levels in urothelial carcinoma of the urinary bladder. Virchows Arch, 2008,452(3):295-304.
[6] Guo F, Cheng X, Jing B, et al. FGD3 binds with HSF4 to suppress p65 expression and inhibit pancreatic cancer progression. Oncogene, 2022,41(6):838-851.
[7] Al-Mutairi M S, Habashy H O. Nuclear factor-κb clinical significance in breast cancer: an immunohistochemical study. Med Princ Pract, 2023,32(1):33-39.
[8] Xu X, Lei Y, Chen L, et al. Phosphorylation of NF-κB p65 drives inflammation-mediated hepatocellular carcinogenesis and is a novel therapeutic target. J Exp Clin Cancer Res, 2021,40(1):253.
[9] Wang S, Kou B, Chai M, et al. Knockout of ASPP2 promotes DEN-induced hepatocarcinogenesis via the NF-kappaB pathway in mice. Cancer Gene Ther, 2022,29(2):202-214.
[10] Hou Y, Moreau F, Chadee K. PPARγ is an E3 ligase that induces the degradation of NFκB/p65. Nat Commun, 2012,3:1300.
[11] Bian X, Liu R, Meng Y, et al. Lipid metabolism and cancer. J Exp Med, 2021,218(1):12-16.
[12] Corn K C, Windham M A, Rafat M. Lipids in the tumor microenvironment: from cancer progression to treatment. Prog Lipid Res, 2020,80:101055.
[13] Chen M, Lu P, Ma Q, et al. CTNNB1/β-catenin dysfunction contributes to adiposity by regulating the cross-talk of mature adipocytes and preadipocytes. Sci Adv, 2020,6(2):eaax9605.
[14] Lawrence T. The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harb Perspect Biol, 2009,1(6):a001651.
[15] Vallabhapurapu S, Karin M. Regulation and function of NF-kappaB transcription factors in the immune system. Annu Rev Immunol, 2009,27:693-733.
[16] Moynagh P N. The NF-kappaB pathway. J Cell Sci, 2005,118(Pt 20):4589-4592.
[17] Liu P, Rojo D L V M, Sammani S, et al. RPA1 binding to NRF2 switches ARE-dependent transcriptional activation to ARE-NRE-dependent repression. Proc Natl Acad Sci U S A, 2018,115(44):E10352-E10361.
[18] Xu M, Tan J, Liu X, et al. Tripartite motif containing 26 prevents steatohepatitis progression by suppressing C/EBPδ signalling activation. Nat Commun, 2023,14(1):6384.
[19] van der Sluis R, Erasmus E. Xenobiotic/medium chain fatty acid: CoA ligase - a critical review on its role in fatty acid metabolism and the detoxification of benzoic acid and aspirin. Expert Opin Drug Metab Toxicol, 2016,12(10):1169-1179.
[20] Cao Y, Li J, Qiu S, et al. ACSM5 inhibits ligamentum flavum hypertrophy by regulating lipid accumulation mediated by FABP4/PPAR signaling pathway. Biol Direct, 2023,18(1):75.
[21] Tsuruta H, Yamahara K, Yasuda-Yamahara M, et al. Emerging pathophysiological roles of ketone bodies. Physiology (Bethesda), 2024,39(3):10.
[22] Shi L, Zhao D, Hou C, et al. Early interleukin-6 enhances hepatic ketogenesis in APP(SWE)/PSEN1dE9 mice via 3-hydroxy-3-methylglutary-CoA synthase 2 signaling activation by p38/nuclear factor κB p65. Neurobiol Aging, 2017,56:115-126.
[23] Asif S, Kim R Y, Fatica T, et al. Hmgcs2-mediated ketogenesis modulates high-fat diet-induced hepatosteatosis. Mol Metab, 2022,61:101494.

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

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