Aberrant acetylated modification of FGF21‑KLB signaling contributes to hepatocellular carcinoma metastasis through the β‑catenin pathway

Xia,Jinkun; Zhu,Zhengyi; Wen,Gaolin; Chen,Yuyan; An,Ran; Xia,Senzhe; Guan,Wenxian; Ren,Haozhen

doi:10.3892/ijo.2023.5539

Aberrant acetylated modification of FGF21‑KLB signaling contributes to hepatocellular carcinoma metastasis through the β‑catenin pathway

Authors:
- Jinkun Xia
- Zhengyi Zhu
- Gaolin Wen
- Yuyan Chen
- Ran An
- Senzhe Xia
- Wenxian Guan
- Haozhen Ren
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Affiliations: Department of Hepatobiliary Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, Jiangsu 210008, P.R. China, Department of General Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Clinical College of Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210008, P.R. China, Hepatobiliary Institute, Medical School, Nanjing University, Nanjing, Jiangsu 210008, P.R. China
Published online on: June 22, 2023 https://doi.org/10.3892/ijo.2023.5539
Article Number: 91

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Abstract

β‑Klotho (KLB) is a vital element of the fibroblast growth factor (FGF) receptor complex and acts as a co‑receptor to facilitate the binding of FGF19 and FGF21 to the FGFRs on the target cells. The present study aimed to determine the contribution of FGF21‑KLB signaling to hepatocellular carcinoma (HCC) metastasis. KLB expression was measured in HCC tissues and cell lines using western blot and reverse transcription‑quantitative PCR. Furthermore, the proliferation, apoptosis and metastasis capacity of KLB‑knockdown Huh7 cells (human HCC cell line) were assessed by Cell Counting Kit‑8 assay, 5‑ethynyl‑2'‑deoxyuridine assay, flow cytometry, wound‑healing assay and Transwell assay. Enrichment analysis was used to explore the underlying regulatory mechanisms of KLB. The metastasis potential of human HCC cells in the context of FGF21 with or without KLB inhibition was determined in vitro and in vivo. Acetylated modification of KLB was determined using a co‑immunoprecipitation assay. The results indicated a significant upregulation of KLB in HCC tissues compared with the corresponding normal tissues. In addition, KLB expression was closely associated with HCC metastasis. Migration and invasion assays revealed that KLB knockdown promoted the metastatic capability of HCC cells. Gene set variation analysis and subsequent mechanistic investigations revealed that KLB is the upstream regulatory factor of β‑catenin signaling. Furthermore, FGF21 was indicated to suppress HCC metastasis by inhibiting β‑catenin signaling‑driven epithelial‑mesenchymal transition (EMT), while KLB knockdown and simultaneous FGF21 overexpression promoted HCC cell motility. Histone deacetylase 3 (HDAC3) was further characterized as the potential deacetylase for KLB. Furthermore, the results revealed that HDAC3 inhibitor‑mediated acetylated modification led to KLB inactivation, resulting in the blockade of FGF21‑KLB signaling, which further triggered the expression of EMT induction‑related genes in Huh7 cells. In conclusion, the present study demonstrated that aberrant acetylated modification of KLB inhibited FGF21‑KLB signaling, thereby promoting β‑catenin signaling‑driven EMT and HCC metastasis.

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1	Llovet JM, Kelley RK, Villanueva A, Singal AG, Pikarsky E, Roayaie S, Lencioni R, Koike K, Zucman-Rossi J and Finn RS: Hepatocellular carcinoma. Nat Rev Dis Primers. 7:62021. View Article : Google Scholar : PubMed/NCBI
2	Biswas S and Rao CM: Epigenetics in cancer: Fundamentals and beyond. Pharmacol Ther. 173:118–134. 2017. View Article : Google Scholar : PubMed/NCBI
3	Kelley RK and Greten TF: Hepatocellular carcinoma-origins and outcomes. N Engl J Med. 385:280–282. 2021. View Article : Google Scholar : PubMed/NCBI
4	Giannelli G, Koudelkova P, Dituri F and Mikulits W: Role of epithelial to mesenchymal transition in hepatocellular carcinoma. J Hepatol. 65:798–808. 2016. View Article : Google Scholar : PubMed/NCBI
5	Ito S, Kinoshita S, Shiraishi N, Nakagawa S, Sekine S, Fujimori T and Nabeshima YI: Molecular cloning and expression analyses of mouse betaklotho, which encodes a novel Klotho family protein. Mech Dev. 98:115–119. 2000. View Article : Google Scholar : PubMed/NCBI
6	Lee S, Choi J, Mohanty J, Sousa LP, Tome F, Pardon E, Steyaert J, Lemmon MA, Lax I and Schlessinger J: Structures of β-klotho reveal a ‘zip code’-like mechanism for endocrine FGF signalling. Nature. 553:501–505. 2018. View Article : Google Scholar : PubMed/NCBI
7	Shi SY, Lu YW, Richardson J, Min X, Weiszmann J, Richards WG, Wang Z, Zhang Z, Zhang J and Li Y: A systematic dissection of sequence elements determining β-Klotho and FGF interaction and signaling. Sci Rep. 8:110452018. View Article : Google Scholar : PubMed/NCBI
8	Triantis V, Saeland E, Bijl N, Oude-Elferink RP and Jansen PL: Glycosylation of fibroblast growth factor receptor 4 is a key regulator of fibroblast growth factor 19-mediated down-regulation of cytochrome P450 7A1. Hepatology. 52:656–666. 2010. View Article : Google Scholar : PubMed/NCBI
9	Huang Z, Xu A and Cheung BMY: The potential role of fibroblast growth factor 21 in lipid metabolism and hypertension. Curr Hypertens Rep. 19:282017. View Article : Google Scholar : PubMed/NCBI
10	Chen J, Du F, Dang Y, Li X, Qian M, Feng W, Qiao C, Fan D, Nie Y, Wu K and Xia L: Fibroblast growth factor 19-mediated up-regulation of SYR-related high-mobility group box 18 promotes hepatocellular carcinoma metastasis by transactivating fibroblast growth factor receptor 4 and Fms-related tyrosine kinase 4. Hepatology. 71:1712–1731. 2020. View Article : Google Scholar : PubMed/NCBI
11	Poh W, Wong W, Ong H, Aung MO, Lim SG, Chua BT and Ho HK: Klotho-beta overexpression as a novel target for suppressing proliferation and fibroblast growth factor receptor-4 signaling in hepatocellular carcinoma. Mol Cancer. 11:142012. View Article : Google Scholar : PubMed/NCBI
12	Anestopoulos I, Voulgaridou GP, Georgakilas AG, Franco R, Pappa A and Panayiotidis MI: Epigenetic therapy as a novel approach in hepatocellular carcinoma. Pharmacol Ther. 145:103–119. 2015. View Article : Google Scholar : PubMed/NCBI
13	Khalili-Tanha G and Moghbeli M: Long non-coding RNAs as the critical regulators of doxorubicin resistance in tumor cells. Cell Mol Biol Lett. 26:392021. View Article : Google Scholar : PubMed/NCBI
14	Han TS, Ban HS, Hur K and Cho HS: The epigenetic regulation of HCC metastasis. Int J Mol Sci. 19:39782018. View Article : Google Scholar : PubMed/NCBI
15	Shen Y, Wei W and Zhou DX: Histone acetylation enzymes coordinate metabolism and gene expression. Trends Plant Sci. 20:614–621. 2015. View Article : Google Scholar : PubMed/NCBI
16	Falkenberg KJ and Johnstone RW: Histone deacetylases and their inhibitors in cancer, neurological diseases and immune disorders. Nat Rev Drug Discov. 13:673–691. 2014. View Article : Google Scholar : PubMed/NCBI
17	Tsilimigras DI, Ntanasis-Stathopoulos I, Moris D, Spartalis E and Pawlik TM: Histone deacetylase inhibitors in hepatocellular carcinoma: A therapeutic perspective. Surg Oncol. 27:611–618. 2018. View Article : Google Scholar : PubMed/NCBI
18	You H, Li Q, Kong D, Liu X, Kong F, Zheng K and Tang R: The interaction of canonical Wnt/β-catenin signaling with protein lysine acetylation. Cell Mol Biol Lett. 27:72022. View Article : Google Scholar : PubMed/NCBI
19	Zhu ZY, Tang N, Wang MF, Zhou JC, Wang JL, Ren HZ and Shi XL: Comprehensive pan-cancer genomic analysis reveals PHF19 as a carcinogenic indicator related to immune infiltration and prognosis of hepatocellular carcinoma. Front Immunol. 12:7810872022. View Article : Google Scholar : PubMed/NCBI
20	Shang L, Ren H, Wang S, Liu H, Hu A, Gou P, Lin Y, Zhou J, Zhu W and Shi X: SS-31 protects liver from ischemia-reperfusion injury via modulating macrophage polarization. Oxid Med Cell Longev. 2021:66621562021. View Article : Google Scholar : PubMed/NCBI
21	Ren HZ, Xia SZ, Qin XQ, Hu AY and Wang JL: FOXO1 alleviates liver ischemia-reperfusion injury by regulating the Th17/Treg ratio through the AKT/Stat3/FOXO1 pathway. J Clin Transl Hepatol. 10:1138–1147. 2022.PubMed/NCBI
22	Wang C, Yang Z, Xu E, Shen X, Wang X, Li Z, Yu H, Chen K, Hu Q, Xia X, et al: Apolipoprotein C-II induces EMT to promote gastric cancer peritoneal metastasis via PI3K/AKT/mTOR pathway. Clin Transl Med. 11:e5222021.PubMed/NCBI
23	Zhang L, Ren CF, Yang Z, Gong LB, Wang C, Feng M and Guan WX: Forkhead Box S1 mediates epithelial-mesenchymal transition through the Wnt/β-catenin signaling pathway to regulate colorectal cancer progression. J Transl Med. 20:3272022. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Zhang P, Martin RC, Cui G, Wang G, Tan Y, Cai L, Lv G and Li Y: Lack of fibroblast growth factor 21 accelerates metabolic liver injury characterized by steatohepatities in mice. Am J Cancer Res. 6:1011–1125. 2016.PubMed/NCBI
25	Suh Y, Yoon CH, Kim RK, Lim EJ, Oh YS, Hwang SG, An S, Yoon G, Gye MC, Yi JM, et al: Claudin-1 induces epithelial-mesenchymal transition through activation of the c-Abl-ERK signaling pathway in human liver cells. Oncogene. 32:4873–4882. 2013. View Article : Google Scholar : PubMed/NCBI
26	Chen F, Gao Q, Wei A, Chen X, Shi Y, Wang H and Cao W: Histone deacetylase 3 aberration inhibits Klotho transcription and promotes renal fibrosis. Cell Death Differ. 28:1001–1012. 2021. View Article : Google Scholar : PubMed/NCBI
27	Sun W, Fu S, Wu S and Tu R: Growing evidence of exosomal MicroRNA-related metastasis of hepatocellular carcinoma. Biomed Res Int. 2020:45014542020. View Article : Google Scholar : PubMed/NCBI
28	Tang W, Chen Z, Zhang W, Cheng Y, Zhang B, Wu F, Wang Q, Wang S, Rong D, Reiter FP, et al: The mechanisms of sorafenib resistance in hepatocellular carcinoma: Theoretical basis and therapeutic aspects. Signal Transduct Target Ther. 5:872020. View Article : Google Scholar : PubMed/NCBI
29	Zhang W, Sun HC, Wang WQ, Zhang QB, Zhuang PY, Xiong YQ, Zhu XD, Xu HX, Kong LQ, Wu WZ, et al: Sorafenib down-regulates expression of HTATIP2 to promote invasiveness and metastasis of orthotopic hepatocellular carcinoma tumors in mice. Gastroenterology. 143:1641–1649.e5. 2012. View Article : Google Scholar : PubMed/NCBI
30	Kim E, Lisby A, Ma C, Lo N, Ehmer U, Hayer KE, Furth EE and Viatour P: Promotion of growth factor signaling as a critical function of β-catenin during HCC progression. Nat Commun. 10:19092019. View Article : Google Scholar : PubMed/NCBI
31	Valenta T, Hausmann G and Basler K: The many faces and functions of β-catenin. EMBO J. 31:2714–2736. 2012. View Article : Google Scholar : PubMed/NCBI
32	Potthoff MJ: FGF21 and metabolic disease in 2016: A new frontier in FGF21 biology. Nat Rev Endocrinol. 13:74–76. 2017. View Article : Google Scholar : PubMed/NCBI
33	Lewis JE, Ebling FJP, Samms RJ and Tsintzas K: Going back to the biology of FGF21: New insights. Trends Endocrinol Metab. 30:491–504. 2019. View Article : Google Scholar : PubMed/NCBI
34	Geng L, Lam KSL and Xu A: The therapeutic potential of FGF21 in metabolic diseases: From bench to clinic. Nat Rev Endocrinol. 16:654–667. 2020. View Article : Google Scholar : PubMed/NCBI
35	Lu W, Li X and Luo Y: FGF21 in obesity and cancer: New insights. Cancer Lett. 499:5–13. 2021. View Article : Google Scholar : PubMed/NCBI
36	Huang X, Yu C, Jin C, Yang C, Xie R, Cao D, Wang F and McKeehan WL: Forced expression of hepatocyte-specific fibroblast growth factor 21 delays initiation of chemically induced hepatocarcinogenesis. Mol Carcinog. 45:934–942. 2006. View Article : Google Scholar : PubMed/NCBI
37	Zheng Q, Martin RC, Shi X, Pandit H, Yu Y, Liu X, Guo W, Tan M, Bai O, Meng X and Li Y: Lack of FGF21 promotes NASH-HCC transition via hepatocyte-TLR4-IL-17A signaling. Theranostics. 10:9923–9936. 2020. View Article : Google Scholar : PubMed/NCBI
38	Liu ZY, Luo Y, Fang AP, Wusiman M, He TT, Liu XZ, Yishake D, Chen S, Lu XT, Zhang YJ and Zhu HL: High serum fibroblast growth factor 21 is associated with inferior hepatocellular carcinoma survival: A prospective cohort study. Liver Int. 42:663–673. 2022. View Article : Google Scholar : PubMed/NCBI
39	Lin YT and Wu KJ: Epigenetic regulation of epithelial-mesenchymal transition: Focusing on hypoxia and TGF-β signaling. J Biomed Sci. 27:392020. View Article : Google Scholar : PubMed/NCBI
40	Hu Y, Zheng Y, Dai M, Wang X, Wu J, Yu B, Zhang H, Cui Y, Kong W, Wu H and Yu X: G9a and histone deacetylases are crucial for Snail2-mediated E-cadherin repression and metastasis in hepatocellular carcinoma. Cancer Sci. 110:3442–3452. 2019. View Article : Google Scholar : PubMed/NCBI
41	Song X, Wang J, Zheng T, Song R, Liang Y, Bhatta N, Yin D, Pan S, Liu J, Jiang H and Liu L: LBH589 inhibits proliferation and metastasis of hepatocellular carcinoma via inhibition of gankyrin/STAT3/Akt pathway. Mol Cancer. 12:1142013. View Article : Google Scholar : PubMed/NCBI
42	Xiao Q, Liu H, Wang HS, Cao MT, Meng XJ, Xiang YL, Zhang YQ, Shu F, Zhang QG, Shan H and Jiang GM: Histone deacetylase inhibitors promote epithelial-mesenchymal transition in hepatocellular carcinoma via AMPK-FOXO1-ULK1 signaling axis-mediated autophagy. Theranostics. 10:10245–10261. 2020. View Article : Google Scholar : PubMed/NCBI
43	Xu W, Liu H, Liu ZG, Wang HS, Zhang F, Wang H, Zhang J, Chen JJ, Huang HJ, Tan Y, et al: Histone deacetylase inhibitors upregulate Snail via Smad2/3 phosphorylation and stabilization of Snail to promote metastasis of hepatoma cells. Cancer Lett. 420:1–13. 2018. View Article : Google Scholar : PubMed/NCBI
44	Ji H, Zhou Y, Zhuang X, Zhu Y, Wu Z, Lu Y, Li S, Zeng Y, Lu QR, Huo Y, et al: HDAC3 deficiency promotes liver cancer through a defect in H3K9ac/H3K9me3 transition. Cancer Res. 79:3676–3688. 2019. View Article : Google Scholar : PubMed/NCBI
45	Cui G, Martin RC, Jin H, Liu X, Pandit H, Zhao H, Cai L, Zhang P, Li W and Li Y: Up-regulation of FGF15/19 signaling promotes hepatocellular carcinoma in the background of fatty liver. J Exp Clin Cancer Res. 37:1362018. View Article : Google Scholar : PubMed/NCBI
46	Kim GY, Kwon JH, Cho JH, Zhang L, Mansfield BC and Chou JY: Downregulation of pathways implicated in liver inflammation and tumorigenesis of glycogen storage disease type Ia mice receiving gene therapy. Hum Mol Genet. 26:1890–1899. 2017. View Article : Google Scholar : PubMed/NCBI