HMGCS2 Mediation of Ketone Levels Affects Sorafenib Treatment Efficacy in Liver Cancer Cells

Primary liver cancer is the fifth leading death of cancers in men, and hepatocellular carcinoma (HCC) accounts for approximately 90% of all primary liver cancer cases. Sorafenib is a first-line drug for advanced-stage HCC patients. Sorafenib is a multi-target kinase inhibitor that blocks tumor cell proliferation and angiogenesis. Despite sorafenib treatment extending survival, some patients experience side effects, and sorafenib resistance does occur. 3-Hydroxymethyl glutaryl-CoA synthase 2 (HMGCS2) is the rate-limiting enzyme for ketogenesis, which synthesizes the ketone bodies, β-hydroxybutyrate (β-HB) and acetoacetate (AcAc). β-HB is the most abundant ketone body which is present in a 4:1 ratio compared to AcAc. Recently, ketone body treatment was found to have therapeutic effects against many cancers by causing metabolic alternations and cancer cell apoptosis. Our previous publication showed that HMGCS2 downregulation-mediated ketone body reduction promoted HCC clinicopathological progression through regulating c-Myc/cyclin D1 and caspase-dependent signaling. However, whether HMGCS2-regulated ketone body production alters the sensitivity of human HCC to sorafenib treatment remains unclear. In this study, we showed that HMGCS2 downregulation enhanced the proliferative ability and attenuated the cytotoxic effects of sorafenib by activating expressions of phosphorylated (p)-extracellular signal-regulated kinase (ERK), p-P38, and p-AKT. In contrast, HMGCS2 overexpression decreased cell proliferation and enhanced the cytotoxic effects of sorafenib in HCC cells by inhibiting ERK activation. Furthermore, we showed that knockdown HMGCS2 exhibited the potential migratory ability, as well as decreasing zonula occludens protein (ZO)-1 and increasing c-Myc expression in both sorafenib-treated Huh7 and HepG2 cells. Although HMGCS2 overexpression did not alter the migratory effect, expressions of ZO-1, c-Myc, and N-cadherin decreased in sorafenib-treated HMGCS2-overexpressing HCC cells. Finally, we investigated whether ketone treatment influences sorafenib sensitivity. We showed that β-HB pretreatment decreased cell proliferation and enhanced antiproliferative effect of sorafenib in both Huh7 and HepG2 cells. In conclusion, this study defined the impacts of HMGCS2 expression and ketone body treatment on influencing the sorafenib sensitivity of liver cancer cells.

[1]  Yuancai Xiang,et al.  Ketogenic diet: new avenues to overcome colorectal cancer , 2022, Signal Transduction and Targeted Therapy.

[2]  S. Siddharth,et al.  Metformin Enhances the Anti-Cancer Efficacy of Sorafenib via Suppressing MAPK/ERK/Stat3 Axis in Hepatocellular Carcinoma , 2022, International journal of molecular sciences.

[3]  Mingyao Li,et al.  β-Hydroxybutyrate suppresses colorectal cancer , 2022, Nature.

[4]  J. Backs,et al.  Ketone body oxidation increases cardiac endothelial cell proliferation , 2022, EMBO molecular medicine.

[5]  A. Rosenzweig,et al.  Ketone bodies for the failing heart: fuels that can fix the engine? , 2021, Trends in Endocrinology & Metabolism.

[6]  Ni Yan,et al.  Local anesthetic bupivacaine inhibits proliferation and metastasis of hepatocellular carcinoma cells via suppressing PI3K/Akt and MAPK signaling , 2021, Journal of biochemical and molecular toxicology.

[7]  Guoqiang Chen,et al.  On the nutritional and therapeutic effects of ketone body d-β-hydroxybutyrate , 2021, Applied Microbiology and Biotechnology.

[8]  S. Ro,et al.  MAPK/ERK Signaling Pathway in Hepatocellular Carcinoma , 2021, Cancers.

[9]  Dana E. Feldman,et al.  Ketogenic diet reduces alcohol withdrawal symptoms in humans and alcohol intake in rodents , 2021, Science Advances.

[10]  F. Papachristou,et al.  Differential effects of cisplatin combined with the flavonoid apigenin on HepG2, Hep3B, and Huh7 liver cancer cell lines. , 2021, Mutation research.

[11]  Yi-Chieh Lee,et al.  Ketogenic Diet Enhances the Cholesterol Accumulation in Liver and Augments the Severity of CCl4 and TAA-Induced Liver Fibrosis in Mice , 2021, International journal of molecular sciences.

[12]  A. Jemal,et al.  Cancer Statistics, 2021 , 2021, CA: a cancer journal for clinicians.

[13]  J. Rungby,et al.  Effects of Ketone Bodies on Brain Metabolism and Function in Neurodegenerative Diseases , 2020, International journal of molecular sciences.

[14]  T. Raife,et al.  β-Hydroxybutyrate inhibits inflammasome activation to attenuate Alzheimer’s disease pathology , 2020, Journal of neuroinflammation.

[15]  H. Maegawa,et al.  SGLT2 Inhibition Mediates Protection from Diabetic Kidney Disease by Promoting Ketone Body-Induced mTORC1 Inhibition. , 2020, Cell metabolism.

[16]  F. Suk,et al.  Loss of HMGCS2 Enhances Lipogenesis and Attenuates the Protective Effect of the Ketogenic Diet in Liver Cancer , 2020, Cancers.

[17]  B. Egan,et al.  Nutritional Ketosis with Ketogenic Diets or Exogenous Ketones: Features, Convergence, and Divergence , 2020, Current sports medicine reports.

[18]  J. Muntané,et al.  Differential effectiveness of tyrosine kinase inhibitors in 2D/3D culture according to cell differentiation, p53 status and mitochondrial respiration in liver cancer cells , 2020, Cell Death & Disease.

[19]  Rajesh Singh,et al.  Liver cancer incidence and mortality: Disparities based on age, ethnicity, health and nutrition, molecular factors, and geography. , 2020, Cancer health disparities.

[20]  W. Walters,et al.  The contribution of ketone bodies to glycolytic inhibition for the treatment of adult and pediatric glioblastoma , 2020, Journal of Neuro-Oncology.

[21]  Sheng-Shou Hu,et al.  Elevated plasma β-hydroxybutyrate predicts adverse outcomes and disease progression in patients with arrhythmogenic cardiomyopathy , 2020, Science Translational Medicine.

[22]  P. Raggi,et al.  The ketogenic diet: Pros and cons. , 2019, Atherosclerosis.

[23]  W. Chiu,et al.  HMGCS2 Mediates Ketone Production and Regulates the Proliferation and Metastasis of Hepatocellular Carcinoma , 2019, Cancers.

[24]  M. Iwano,et al.  β-Hydroxybutyrate enhances the cytotoxic effect of cisplatin via the inhibition of HDAC/survivin axis in human hepatocellular carcinoma cells. , 2019, Journal of pharmacological sciences.

[25]  Guangji Wang,et al.  Apatinib induces 3‐hydroxybutyric acid production in the liver of mice by peroxisome proliferator‐activated receptor α activation to aid its antitumor effect , 2019, Cancer science.

[26]  Gregory J. Gores,et al.  A global view of hepatocellular carcinoma: trends, risk, prevention and management , 2019, Nature Reviews Gastroenterology & Hepatology.

[27]  Y. Liu,et al.  Identification of β-hydroxybutyrate as a potential biomarker for female papillary thyroid cancer. , 2019, Bioanalysis.

[28]  M. Kudo,et al.  Lenvatinib versus sorafenib in first-line treatment of patients with unresectable hepatocellular carcinoma: a randomised phase 3 non-inferiority trial , 2018, The Lancet.

[29]  K. Clarke,et al.  A Ketone Ester Drink Lowers Human Ghrelin and Appetite , 2017, Obesity.

[30]  Shu-Guang Su,et al.  miR-107-mediated decrease of HMGCS2 indicates poor outcomes and promotes cell migration in hepatocellular carcinoma. , 2017, The international journal of biochemistry & cell biology.

[31]  Caifeng Xie,et al.  The role of OXCT1 in the pathogenesis of cancer as a rate‐limiting enzyme of ketone body metabolism , 2017, Life sciences.

[32]  D. Scholtens,et al.  Overexpression of lipid metabolism genes and PBX1 in the contralateral breasts of women with estrogen receptor‐negative breast cancer , 2017, International journal of cancer.

[33]  J. Dietrich,et al.  Role of ketogenic metabolic therapy in malignant glioma: A systematic review. , 2017, Critical reviews in oncology/hematology.

[34]  Lei Chen,et al.  New knowledge of the mechanisms of sorafenib resistance in liver cancer , 2017, Acta Pharmacologica Sinica.

[35]  King-Jen Chang,et al.  Adipocytes promote malignant growth of breast tumours with monocarboxylate transporter 2 expression via β-hydroxybutyrate , 2017, Nature Communications.

[36]  J. Griffiths,et al.  The action of β-hydroxybutyrate on the growth, metabolism and global histone H3 acetylation of spontaneous mouse mammary tumours: evidence of a β-hydroxybutyrate paradox , 2017, Cancer & metabolism.

[37]  P. Puchalska,et al.  Multi-dimensional Roles of Ketone Bodies in Fuel Metabolism, Signaling, and Therapeutics. , 2017, Cell metabolism.

[38]  K. Reiss,et al.  Regulation of Ketone Body Metabolism and the Role of PPARα , 2016, International journal of molecular sciences.

[39]  Petra Koudelkova,et al.  Role of epithelial to mesenchymal transition in hepatocellular carcinoma. , 2016, Journal of hepatology.

[40]  P. Asih,et al.  Differential expression of several drug transporter genes in HepG2 and Huh-7 cell lines , 2016, Advanced biomedical research.

[41]  A. Wilk,et al.  Fenofibrate Induces Ketone Body Production in Melanoma and Glioblastoma Cells , 2016, Front. Endocrinol..

[42]  K. Hui,et al.  EDIL3 is a novel regulator of epithelial-mesenchymal transition controlling early recurrence of hepatocellular carcinoma. , 2015, Journal of hepatology.

[43]  Ting-Feng Wu,et al.  The prognostic impact of lipid biosynthesis-associated markers, HSD17B2 and HMGCS2, in rectal cancer treated with neoadjuvant concurrent chemoradiotherapy , 2015, Tumor Biology.

[44]  G. Song,et al.  Sorafenib inhibits migration and invasion of hepatocellular carcinoma cells through suppression of matrix metalloproteinase expression. , 2015, Anticancer research.

[45]  T. Vanitallie,et al.  Ketone body therapy: from the ketogenic diet to the oral administration of ketone ester , 2014, Journal of Lipid Research.

[46]  R. Powers,et al.  Metabolic reprogramming induced by ketone bodies diminishes pancreatic cancer cachexia , 2014, Cancer & metabolism.

[47]  Kuen-Feng Chen,et al.  SC-2001 Overcomes STAT3-mediated Sorafenib Resistance through RFX-1/SHP-1 Activation in Hepatocellular Carcinoma , 2014, Neoplasia.

[48]  T. Seyfried,et al.  Ketone supplementation decreases tumor cell viability and prolongs survival of mice with metastatic cancer , 2014, International journal of cancer.

[49]  Brian J. Smith,et al.  Ketogenic Diets Enhance Oxidative Stress and Radio-Chemo-Therapy Responses in Lung Cancer Xenografts , 2013, Clinical Cancer Research.

[50]  T. H. van der Kwast,et al.  Quantitative Proteomics Reveals That Enzymes of the Ketogenic Pathway Are Associated with Prostate Cancer Progression* , 2013, Molecular & Cellular Proteomics.

[51]  Kuen-Feng Chen,et al.  Activation of Phosphatidylinositol 3-Kinase/Akt Signaling Pathway Mediates Acquired Resistance to Sorafenib in Hepatocellular Carcinoma Cells , 2011, Journal of Pharmacology and Experimental Therapeutics.

[52]  M. Kudo,et al.  Sorafenib Inhibits the Hepatocyte Growth Factor–Mediated Epithelial Mesenchymal Transition in Hepatocellular Carcinoma , 2011, Molecular Cancer Therapeutics.

[53]  W. Yue,et al.  Crystal structures of human HMG-CoA synthase isoforms provide insights into inherited ketogenesis disorders and inhibitor design. , 2010, Journal of molecular biology.

[54]  Robert E. Brown,et al.  Sorafenib downregulates ERK/Akt and STAT3 survival pathways and induces apoptosis in a human neuroblastoma cell line. , 2010, International journal of clinical and experimental pathology.

[55]  Gary Yellen,et al.  Ketone bodies, glycolysis, and KATP channels in the mechanism of the ketogenic diet , 2008, Epilepsia.

[56]  R. Wittig,et al.  Growth of human gastric cancer cells in nude mice is delayed by a ketogenic diet supplemented with omega-3 fatty acids and medium-chain triglycerides , 2008, BMC Cancer.

[57]  C. Liang,et al.  In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro , 2007, Nature Protocols.

[58]  D. Haro,et al.  Ketogenic HMGCS2 Is a c-Myc Target Gene Expressed in Differentiated Cells of Human Colonic Epithelium and Down-Regulated in Colon Cancer , 2006, Molecular Cancer Research.

[59]  E. Nishida,et al.  ERK MAP kinase in G1 cell cycle progression and cancer , 2006, Cancer science.

[60]  R. DeMatteo,et al.  Molecular mechanisms in hepatocellular carcinoma development. , 2005, Best practice & research. Clinical gastroenterology.

[61]  W. H. Porter,et al.  Laboratory and clinical evaluation of assays for β-hydroxybutyrate , 1997 .

[62]  W. H. Porter,et al.  Laboratory and clinical evaluation of assays for beta-hydroxybutyrate. , 1997, American journal of clinical pathology.

[63]  F. Hegardt,et al.  Molecular cloning and tissue expression of human mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase. , 1995, Archives of biochemistry and biophysics.