Extracellular Citrate Treatment Induces HIF1α Degradation and Inhibits the Growth of Low-Glycolytic Hepatocellular Carcinoma under Hypoxia
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Human HCC Samples
2.2. Cell Culture
2.3. In Vitro 14C-Deoxyglucose (DG) and 14C-Citrate Uptake Assay
2.4. Cell Viability and ATP Measurement
2.5. Western Blotting
2.6. Flow Cytometry
2.7. In Vivo Experiment Using HepG2-Xenograft Animal Model
2.8. RNA Expression
2.9. Statistical Analysis
3. Results
3.1. Correlation of SLC-Related Genes and Proteins in HCC Patients with High- and Low-Glycolysis
3.2. Citrate Treatment Affects HIF1α in HCC Cells with High NaCT Expression
3.3. Effects of Citrate Treatment on Glycolytic Pathway and HIF1α Regulation
3.4. Anti-Tumor Effects of Citrate in Human HCC Cell Lines and Mouse Xenograft Models
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Sung, H.; Ferlay, J.; Siegel, R.L.; Laversanne, M.; Soerjomataram, I.; Jemal, A.; Bray, F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. Ca-Cancer J. Clin. 2021, 71, 209–249. [Google Scholar] [CrossRef] [PubMed]
- Hwang, S.H.; Lee, M.; Lee, N.; Park, S.; Kim, C.K.; Park, M.A.; Yun, M. Increased 18F-FDG uptake on PET/CT is associated with poor arterial and portal perfusion on multiphase CT. Clin. Nucl. Med. 2016, 41, 296–301. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.; Stewart, E.; Desjardins, L.; Hadway, J.; Morrison, L.; Crukley, C.; Lee, T.Y. Assessment of intratumor hypoxia by integrated 18F-FDG-PET / perfusion CT in a liver tumor model. PLoS ONE 2017, 12, e0173016. [Google Scholar] [CrossRef]
- Zhang, L.Y.; Li, Y.; Dai, Y.B.; Wang, D.H.; Wang, X.C.; Cao, Y.; Liu, W.W.; Tao, Z.H. Glycolysis-related gene expression profiling serves as a novel prognosis risk predictor for human hepatocellular carcinoma. Sci. Rep. 2021, 11, 1875. [Google Scholar] [CrossRef] [PubMed]
- Lee, N.C.W.; Carella, M.A.; Papa, S.; Bubici, C. High expression of glycolytic genes in cirrhosis correlates with the risk of developing liver cancer. Front. Cell Dev. Biol. 2018, 6, 138. [Google Scholar] [CrossRef] [Green Version]
- Chen, H.; Wu, Q.; Peng, L.; Cao, T.; Deng, M.L.; Liu, Y.W.; Huang, J.; Hu, Y.; Fu, N.; Zhou, K.B.; et al. Mechanism, Clinical significance, and treatment strategy of Warburg effect in hepatocellular carcinoma. J. Nanomater. 2021, 2021, 5164100. [Google Scholar] [CrossRef]
- Zhu, J.; Thompson, C.B. Metabolic regulation of cell growth and proliferation. Nat. Rev. Mol. Cell Biol. 2019, 20, 436–450. [Google Scholar] [CrossRef]
- Zhang, Y.; Zhang, Y.; Sun, K.; Meng, Z.; Chen, L. The SLC transporter in nutrient and metabolic sensing, regulation, and drug development. J. Mol. Cell Biol. 2019, 11, 1–13. [Google Scholar] [CrossRef] [Green Version]
- Yin, C.Q.; Qie, S.; Sang, N.L. Carbon source metabolism and its regulation in cancer cells. Crit. Rev. Eukar. Gene 2012, 22, 17–35. [Google Scholar] [CrossRef] [Green Version]
- Liberti, M.V.; Locasale, J.W. The Warburg effect: How does it benefit cancer cells? Trends Biochem. Sci. 2016, 41, 287. [Google Scholar] [CrossRef]
- Keenan, M.M.; Chi, J.T. Alternative Fuels for cancer cells. Cancer J. 2015, 21, 49–55. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jeon, J.Y.; Lee, M.; Whang, S.H.; Kim, J.W.; Cho, A.; Yun, M. Regulation of acetate utilization by monocarboxylate transporter 1 (MCT1) in hepatocellular carcinoma (HCC). Oncol. Res. 2018, 26, 71–81. [Google Scholar] [CrossRef] [PubMed]
- Yun, M.; Bang, S.H.; Kim, J.W.; Park, J.Y.; Kim, K.S.; Lee, J.D. The importance of acetyl coenzyme A synthetase for C-11-acetate uptake and cell survival in hepatocellular carcinoma. J. Nucl. Med. 2009, 50, 1222–1228. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Schug, Z.T.; Vande Voorde, J.; Gottlieb, E. The metabolic fate of acetate in cancer. Nat. Rev. Cancer 2016, 16, 708–717. [Google Scholar] [CrossRef]
- Ho, C.L.; Yu, S.C.; Yeung, D.W. 11C-acetate PET imaging in hepatocellular carcinoma and other liver masses. J. Nucl. Med. 2003, 44, 213–221. [Google Scholar]
- Feron, O. The many metabolic sources of Acetyl-CoA to support histone acetylation and influence cancer progression. Ann. Transl. Med. 2019, 7 (Suppl. S8), S277. [Google Scholar] [CrossRef]
- Martinez-Reyes, I.; Chandel, N.S. Mitochondrial TCA cycle metabolites control physiology and disease. Nat. Commun. 2020, 11, 102. [Google Scholar] [CrossRef] [Green Version]
- Fendt, S.M.; Bell, E.L.; Keibler, M.A.; Olenchock, B.A.; Mayers, J.R.; Wasylenko, T.M.; Vokes, N.I.; Guarente, L.; Vander Heiden, M.G.; Stephanopoulos, G. Reductive glutamine metabolism is a function of the alpha-ketoglutarate to citrate ratio in cells. Nat. Commun. 2013, 4, 2236. [Google Scholar] [CrossRef] [Green Version]
- Mycielska, M.E.; Dettmer, K.; Rummele, P.; Schmidt, K.; Prehn, C.; Milenkovic, V.M.; Jagla, W.; Madej, G.M.; Lantow, M.; Schladt, M.; et al. Extracellular Citrate Affects Critical Elements o Cancer Cell Metabolism and Supports Cancer Development In Vivo. Cancer Res. 2018, 78, 2513–2523. [Google Scholar] [CrossRef] [Green Version]
- Iacobazzi, V.; Infantino, V. Citrate—New functions for an old metabolite. Biol. Chem. 2014, 395, 387–399. [Google Scholar] [CrossRef]
- Haferkamp, S.; Drexler, K.; Federlin, M.; Schlitt, H.J.; Berneburg, M.; Adamski, J.; Gaumann, A.; Geissler, E.K.; Ganapathy, V.; Parkinson, E.L.; et al. Extracellular citrate fuels cancer cell metabolism and growth. Front. Cell Dev. Biol. 2020, 8, 602476. [Google Scholar] [CrossRef] [PubMed]
- Ren, J.G.; Seth, P.; Ye, H.H.; Guo, K.; Hanai, J.; Husain, Z.; Sukhatme, V.P. Citrate suppresses tumor growth in multiple models through inhibition of glycolysis, the tricarboxylic acid cycle and the IGF-1R pathway. Sci. Rep. 2017, 7, 4537. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wang, T.A.; Zhang, X.D.; Guo, X.Y.; Xian, S.L.; Lu, Y.F. 3-bromopyruvate and sodium citrate target glycolysis suppress surviving, and induce mitochondrial-mediated apoptosis in gastric cancer cells and inhibit gastric orthotopic transplantation tumor growth. Oncol. Rep. 2016, 35, 1287–1296. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lu, Y.; Zhang, X.; Ahang, H.; Lan, J.; Huang, G.; Varin, E.; Lincet, H.; Poulain, L.; Icard, P. Citrate induces apoptotic cell death: A promising way to treat gastric carcinoma? Anticancer Res. 2011, 31, 797–805. [Google Scholar]
- Yan, Q.Y.; Lu, Y.T.; Zhou, L.L.; Chen, J.L.; Xu, H.J.; Cai, M.J.; Shi, Y.; Jiang, J.G.; Xiong, W.Y.; Gao, J.; et al. Mechanistic insights into GLUT1 activation and clustering revealed by supper-resolution imaging. Proc. Natl. Acad. Sci. USA 2018, 115, 7033–7038. [Google Scholar] [CrossRef] [Green Version]
- Joshi, S.; Singh, A.R.; Durden, D.L. MDM2 regulates hypoxic hypoxia-inducible factor 1alpha stability in an E3 ligase, proteasome, and PETN-phosphatidylinositol 3-kinase-AKT dependent manner. J. Biol. Chem. 2014, 289, 22785–22797. [Google Scholar] [CrossRef] [Green Version]
- Li, Y.; Zschaeck, S.; Liq, Q.; Chen, S.; Chen, L.; Wu, H. Metabolic parameters of sequential 18F-FDG PET/CT predict overall survival of esophageal cancer patients treated with (chemo-) radiation. Radiat. Oncol. 2019, 14, 35. [Google Scholar] [CrossRef]
- Li, X.F.; Du, Y.; Ma, Y.Y.; Postel, G.C.; Civelek, A.C. F-18-Fluorodeoxyglucose uptake and tumor hypoxia: Revisit F-18-fluorodexyglucose in oncology application. Transl. Oncol. 2014, 7, 240–247. [Google Scholar] [CrossRef] [Green Version]
- Ganapathy, V.; Thangaraju, M.; Prasad, P.D. Nutrient transporters in cancer: Relevance to Warburg hypothesis and beyond. Pharmacol. Ther. 2009, 121, 29–40. [Google Scholar] [CrossRef]
- Cassim, S.; Raymond, V.A.; Dehbidi-Assadzadeh, L.; Lapierre, P.; Bilodeau, M. Metabolic reprogramming enables hepatocarcinoma cells to efficiently adapt and survive to a nutrient-restricted microenvironment. Cell Cycle 2018, 17, 903–916. [Google Scholar] [CrossRef]
- Comerford, S.A.; Huang, Z.; Du, X.; Wang, Y.; Cai, L.; Witkiewicz, A.K.; Walters, H.; Tantawy, M.N.; Fu, A.; Manning, H.C.; et al. Acetate dependence of tumors. Cell 2014, 159, 1591–1602. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lee, M.; Jeon, J.Y.; Neugent, M.L.; Kim, J.W.; Yun, M. 18F-fluorodeoxyglucose uptake on positron emission tomography/computed tomography is associated with metastasis and epithelial-mesenchymal transition in hepatocellular carcinoma. Clin. Exp. Metastasis 2017, 34, 251–260. [Google Scholar] [CrossRef] [PubMed]
- Park, J.W.; Kim, J.H.; Kim, S.K.; Kang, K.W.; Park, K.W.; Choi, J.I.; Lee, W.J.; Kim, C.M.; Nam, B.H. A prospective evaluation of 18F-FDG and 11C-acetate PET/CT for detection of primary and metastatic hepatocellular carcinoma. J. Nucl. Med. 2008, 49, 1912–1921. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kopel, J.; Higuchi, K.; Ristic, B.; Sato, T.; Ramachandran, S.; Ganapathy, V. The hepatic plasma membrane citrate transporter NaCT (SLC13A5) as a molecular target for metformin. Sci. Rep. 2020, 10, 8536. [Google Scholar] [CrossRef] [PubMed]
- Iommarini, L.; Porcelli, A.M.; Gasparre, G.; Kurelac, I. Non-canonical mechanisms regulating hypoxia-inducible factor 1 alpha in cancer. Front. Oncol. 2017, 7, 286. [Google Scholar] [CrossRef] [Green Version]
- Urso, L.; Calabrese, F.; Faveretto, A.; Conte, P.; Pasello, G. Critical review about MDM2 in cancer: Possible role in malignant mesothelioma and implications for treatment. Crit. Rev. Oncol. Hematol. 2016, 97, 220–230. [Google Scholar] [CrossRef]
- Lim, J.H.; Lee, Y.M.; Chun, Y.S.; Chen, J.; Kim, J.E.; Park, J.W. Sirtuin 1 modulates cellular responses to hypoxia by deacetylating hypoxia-inducible factor 1 alpha. Mol. Cell 2010, 38, 864–878. [Google Scholar] [CrossRef]
- Joo, H.Y.; Yun, M.; Jeong, J.; Park, E.R.; Shin, H.J.; Woo, S.R.; Jung, J.K.; Kim, Y.M.; Park, J.J.; Kim, J.; et al. SIRT1 deacetylates and stabilizes hypoxia-inducible factor-1 alpha (HIF-1 alpha) via direct interactions during hypoxia. Biochem. Bioph. Res. Commun. 2015, 462, 294–300. [Google Scholar] [CrossRef]
- Halabe Bucay, A. Hypothesis proved citric acid (citrate) does improve cancer: A case of a patient suffering from medullary thyroid cancer. Med. Hypotheses 2009, 73, 271. [Google Scholar] [CrossRef]
- Bucay, A.H. Clinical report: A patient with primary peritoneal mesothelioma that has improved after taking citric acid orally. Clin. Res. Hepatol. Gas 2011, 35, 241. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Kim, S.Y.; Kim, D.; Kim, J.; Ko, H.Y.; Kim, W.J.; Park, Y.; Lee, H.W.; Han, D.H.; Kim, K.S.; Park, S.; et al. Extracellular Citrate Treatment Induces HIF1α Degradation and Inhibits the Growth of Low-Glycolytic Hepatocellular Carcinoma under Hypoxia. Cancers 2022, 14, 3355. https://doi.org/10.3390/cancers14143355
Kim SY, Kim D, Kim J, Ko HY, Kim WJ, Park Y, Lee HW, Han DH, Kim KS, Park S, et al. Extracellular Citrate Treatment Induces HIF1α Degradation and Inhibits the Growth of Low-Glycolytic Hepatocellular Carcinoma under Hypoxia. Cancers. 2022; 14(14):3355. https://doi.org/10.3390/cancers14143355
Chicago/Turabian StyleKim, Seon Yoo, Dongwoo Kim, Jisu Kim, Hae Young Ko, Won Jin Kim, Youngjoo Park, Hye Won Lee, Dai Hoon Han, Kyung Sik Kim, Sunghyouk Park, and et al. 2022. "Extracellular Citrate Treatment Induces HIF1α Degradation and Inhibits the Growth of Low-Glycolytic Hepatocellular Carcinoma under Hypoxia" Cancers 14, no. 14: 3355. https://doi.org/10.3390/cancers14143355