The mechanism of histone deacetylase inhibitor vorinostat inhibiting the growth of hepatocellular carcinoma cells

Abstract

Abstract: Objective: To investigate the mechanism of exonuclease 1 (EXO1) in hepatocellular carcinoma (HCC) and the possible mechanism of the histone deacetylase (HDAC) inhibitor vorinostat inhibiting the growth of HCC cells.
Methods: HCC cell lines with knockdown or overexpression of EXO1 were constructed. And the effects of EXO1 on the proliferation, migration and invasion in HCC were investigated by cell phenotype assays. The role of EXO1 in homologous recombination (HR) repair and its effect on ionizing radiation sensitivity of cells were detected by HR reportor assay, immunofluorescence and western blots. Additionally, HCC cells were treated with vorinostat. The effect of vorinostat on EXO1 expression and the possible mechanism of inhibiting the growth of HCC cells were detected by CCK-8, flow cytometry and Western blotting.
Results: High level of EXO1 promoted the proliferation, migration and invasion of HCC cells. EXO1 also promoted DNA double-strand break repair through the HR pathway and reduced cell sensitivity to ionizing radiation. In addition, elevated EXO1 levels were less sensitive to vorinostat, whereas knockdown of EXO1 increased the sensitivity of hepatocellular carcinoma cells to vorinostat. Vorinostat further inhibited the HR repair ability of HCC cells by suppressing EXO1 expression, thereby inhibiting cell growth.
Conclusion: Vorinostat can inhibit HR repair ability by decreasing the expression level of EXO1, which may serve as a potential therapeutic target for HCC

Key words: Exonuclease 1, Hepatocellular carcinoma, Vorinostat, Homologous recombination repair

Gu Xinglu, Zhao Xiaohang, Sun Yulin. The mechanism of histone deacetylase inhibitor vorinostat inhibiting the growth of hepatocellular carcinoma cells[J]. Electronic Journal of Liver Tumor, 2023, 10(1): 15-24.

References

[1] SUNG H, FERLAY J, SIEGEL R L, et al.Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249.
[2] VILLANUEVA A.Hepatocellular Carcinoma[J]. N Engl J Med, 2019, 380(15): 1450-1462.
[3] KRISHNAMURTHY S, GILOT D, AHN S B, et al.Involvement of Kynurenine Pathway in Hepatocellular Carcinoma[J]. Cancers (Basel), 2021, 13(20): 5180.
[4] ANWANWAN D, SINGH S K, SINGH S, et al.Challenges in liver cancer and possible treatment approaches[J]. Biochim Biophys Acta Rev Cancer, 2020, 1873(1): 188314.
[5] LIU D, SONG T.Changes in and challenges regarding the surgical treatment of hepatocellular carcinoma in China[J]. Biosci Trends, 2021, 15(3): 142-147.
[6] XU J, JI L, RUAN Y, et al.UBQLN1 mediates sorafenib resistance through regulating mitochondrial biogenesis and ROS homeostasis by targeting PGC1beta in hepatocellular carcinoma[J]. Signal Transduct Target Ther, 2021, 6(1): 190.
[7] SINGH A K, SINGH S V, KUMAR R, et al.Current therapeutic modalities and chemopreventive role of natural products in liver cancer: Progress and promise[J]. World J Hepatol, 2023, 15(1): 1-18.
[8] GUO J, ZHAO J, XU Q, et al.Resistance of Lenvatinib in Hepatocellular Carcinoma[J]. Curr Cancer Drug Targets, 2022, 22(11): 865-878.
[9] LIU K Y, WANG L T, HSU S H.Modification of Epigenetic Histone Acetylation in Hepatocellular Carcinoma[J]. Cancers, 2018, 10(1): 8.
[10] DU G, YANG R, QIU J, et al.Multifaceted Influence of Histone Deacetylases on DNA Damage Repair: Implications for Hepatocellular Carcinoma[J]. J Clin Transl Hepatol, 2023, 11(1): 231-243.
[11] ALCITEPE I, SALCIN H, KARATEKIN I, et al.HDAC inhibitor Vorinostat and BET inhibitor Plx51107 epigenetic agents' combined treatments exert a therapeutic approach upon acute myeloid leukemia cell model[J]. Med Oncol, 2022, 39(12): 257.
[12] SANAEI M, KAVOOSI F, POURAHMADI M.Effect of Decitabine (5-aza-2'-deoxycytidine, 5-aza-CdR) in Comparison with Vorinostat (Suberoylanilide Hydroxamic Acid, SAHA) on DNMT1, DNMT3a and DNMT3b, HDAC 1-3, SOCS 1, SOCS 3, JAK2, and STAT3 Gene Expression in Hepatocellular Carcinoma HLE and LCL-PI 11 Cell Lines[J]. Asian Pac J Cancer Prev, 2021, 22(7): 2089-2098.
[13] SANAEI M, KAVOOSI F, ESMI Z.The Effect of 5-Aza-2'-Deoxycytidine in Combination to and in Comparison with Vorinostat on DNA Methyltransferases, Histone Deacetylase 1, Glutathione S-Transferase 1 and Suppressor of Cytokine Signaling 1 Genes Expression, Cell Growth Inhibition and Apoptotic Induction in Hepatocellular LCL-PI 11 Cell Line[J]. Int J Hematol Oncol Stem Cell Res, 2020, 14(1): 45-55.
[14] KEIJZERS G, BAKULA D, PETR M A, et al.Human Exonuclease 1 (EXO1) Regulatory Functions in DNA Replication with Putative Roles in Cancer[J]. Int J Mol Sci, 2018, 20(1): 74.
[15] LUO F, WANG Y Z, LIN D, et al.Exonuclease 1 expression is associated with clinical progression, metastasis, and survival prognosis of prostate cancer[J]. J Cell Biochem, 2019, 120(7): 11383-11389.
[16] SHI Q, YAO X Y, WANG H Y, et al.Breast cancer-associated SNP rs72755295 is a cis-regulatory variation for human EXO1[J]. Genet Mol Biol, 2022, 45(4): e20210420.
[17] ZHOU C S, FENG M T, CHEN X, et al.Exonuclease 1 (EXO1) is a Potential Prognostic Biomarker and Correlates with Immune Infiltrates in Lung Adenocarcinoma[J]. Onco Targets Ther, 2021, 14(1): 1033-1048.
[18] ALIMIRZAIE S, MOHAMADKHANI A, MASOUDI S, et al.Mutations in Known and Novel cancer Susceptibility Genes in Young Patients with Pancreatic Cancer[J]. Arch Iran Med, 2018, 21(6): 228-233.
[19] MAO P, WU S, FAN Y.Upregulation of exonuclease 1 caused by homology-dependent repair confers cisplatin resistance to gastric cancer cells[J]. Can J Physiol Pharmacol, 2022, 100(9): 903-914.
[20] CHEN J, WANG Z, SHEN X, et al.Identification of novel biomarkers and small molecule drugs in human colorectal cancer by microarray and bioinformatics analysis[J]. Mol Genet Genomic Med, 2019, 7(7): e00713.
[21] DAI Y, TANG Z, YANG Z, et al.EXO1 overexpression is associated with poor prognosis of hepatocellular carcinoma patients[J]. Cell Cycle, 2018, 17(19-20): 2386-2397.
[22] WANG Z L, LI B, LUO Y X, et al.Comprehensive Genomic Characterization of RNA-Binding Proteins across Human Cancers[J]. Cell Rep, 2018, 22(1): 286-98.
[23] YANG G, DONG K, ZHANG Z, et al.EXO1 Plays a Carcinogenic Role in Hepatocellular Carcinoma and is related to the regulation of FOXP3[J]. J Cancer, 2020, 11(16): 4917-32.
[24] MA J, JIN J, LU H, et al.Exonuclease 1 is a Potential Diagnostic and Prognostic Biomarker in Hepatocellular Carcinoma[J]. Front Mol Biosci, 2022, 9(1): 889414.
[25] XU Y, XU D.Repair pathway choice for double-strand breaks[J]. Essays Biochem, 2020, 64(5): 765-777.
[26] ZHAO F, KIM W, KLOEBER J A, et al.DNA end resection and its role in DNA replication and DSB repair choice in mammalian cells[J]. Exp Mol Med, 2020, 52(10): 1705-1714.
[27] LI Y, WANG Y, ZHANG W, et al.BKM120 sensitizes BRCA-proficient triple negative breast cancer cells to olaparib through regulating FOXM1 and Exo1 expression[J]. Sci Rep, 2021, 11(1): 4774.
[28] MIRICESCU D, TOTAN A, STANESCU-SPINU I I, et al. PI3K/AKT/mTOR Signaling Pathway in Breast Cancer: From Molecular Landscape to Clinical Aspects[J]. Int J Mol Sci, 2020, 22(1): 173.
[29] MUTHUSWAMI M, RAMESH V, BANERJEE S, et al.Breast tumors with elevated expression of 1q candidate genes confer poor clinical outcome and sensitivity to Ras/PI3K inhibition[J]. PLoS One, 2013, 8(10): e77553.
[30] TANG D, HUANG T, TIAN Q, et al.MYC/NBS1-Mediated DNA Damage Response is Involved in the Inhibitory Effect of Hydroxysafflor Yellow A on Glioma Cells[J]. Drug Des Devel Ther, 2021, 15: 1749-1763.
[31] VANAN I, DONG Z, TOSTI E, et al.Role of a DNA damage checkpoint pathway in ionizing radiation-induced glioblastoma cell migration and invasion[J]. Cell Mol Neurobiol, 2012, 32(7): 1199-1208.
[32] LEE J O, KANG M J, BYUN W S, et al.Metformin overcomes resistance to cisplatin in triple-negative breast cancer (TNBC) cells by targeting RAD51[J]. Breast Cancer Res, 2019, 21(1): 115.
[33] LIAO B, ZHANG Y, SUN Q, et al.Vorinostat enhances the anticancer effect of oxaliplatin on hepatocellular carcinoma cells[J]. Cancer Med, 2018, 7(1): 196-207.
[34] LI Y L, RAO M J, ZHANG N Y, et al.BAY 87-2243 sensitizes hepatocellular carcinoma Hep3B cells to histone deacetylase inhibitors treatment via GSK-3beta activation[J]. Exp Ther Med, 2019, 17(6): 4547-4553.
[35] BOTRUGNO O A, ROBERT T, VANOLI F, et al.Molecular pathways: old drugs define new pathways: non-histone acetylation at the crossroads of the DNA damage response and autophagy[J]. Clin Cancer Res, 2012, 18(9): 2436-2442.
[36] MUNSHI A, TANAKA T, HOBBS M L, et al.Vorinostat, a histone deacetylase inhibitor, enhances the response of human tumor cells to ionizing radiation through prolongation of gamma-H2AX foci[J]. Mol Cancer Ther, 2006, 5(8): 1967-1974.