Inhibition of Heme Export and/or Heme Synthesis Potentiates Metformin Anti-Proliferative Effect on Cancer Cell Lines
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Gene Silencing and Overexpression
2.3. RNA Extraction and Quantitative Real-Time-PCR Analysis
2.4. Western Blot
2.5. Cells Treatment with Drugs
2.6. Evaluation of Cell Proliferation
2.7. Crystal Violet Staining
2.8. Mitochondria Isolation and ETC Complexes Activity Measurment
2.9. Statistical Analyses
3. Results
3.1. FLVCR1a Silencing Improves Metformin Effect on CRC Cell Lines
3.2. ALA Treatment Improves Metformin Effect on CRC Cell Lines
3.3. The Down-Modulation of the Heme Synthesis-Export System Improves Metformin Effect on Different Kinds of Tumor Cell Lines
4. Discussion
5. Conclusions
6. Patents
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Chiabrando, D.; Mercurio, S.; Tolosano, E. Heme and erythropoieis: More than a structural role. Haematologica 2014, 99, 973–983. [Google Scholar] [CrossRef]
- Fiorito, V.; Chiabrando, D.; Petrillo, S.; Bertino, F.; Tolosano, E. The Multifaceted Role of Heme in Cancer. Front. Oncol. 2020, 9, 1540. [Google Scholar] [CrossRef] [Green Version]
- Chiabrando, D.; Vinchi, F.; Fiorito, V.; Mercurio, S.; Tolosano, E. Heme in pathophysiology: A matter of scavenging, metabolism and trafficking across cell membranes. Front. Pharmacol. 2014, 5, 61. [Google Scholar] [CrossRef] [Green Version]
- Krishnamurthy, P.; Schuetz, J.D. The role of ABCG2 and ABCB6 in porphyrin metabolism and cell survival. Curr. Pharm. Biotechnol. 2011, 12, 647–655. [Google Scholar] [CrossRef]
- Quigley, J.G.; Yang, Z.; Worthington, M.T.; Phillips, J.D.; Sabo, K.M.; Sabath, D.E.; Berg, C.L.; Sassa, S.; Wood, B.L.; Abkowitz, J.L. Identification of a human heme exporter that is essential for erythropoiesis. Cell 2004, 118, 757–766. [Google Scholar] [CrossRef] [Green Version]
- Fiorito, V.; Allocco, A.L.; Petrillo, S.; Gazzano, E.; Torretta, S.; Marchi, S.; Destefanis, F.; Pacelli, C.; Audrito, V.; Provero, P.; et al. The heme synthesis-export system regulates the tricarboxylic acid cycle flux and oxidative phosphorylation. Cell Rep. 2021, 35, 109252. [Google Scholar] [CrossRef]
- Lee, J.; Yesilkanal, A.E.; Wynne, J.P.; Frankenberger, C.; Liu, J.; Yan, J.; Elbaz, M.; Rabe, D.C.; Rustandy, F.D.; Tiwari, P.; et al. Effective breast cancer combination therapy targeting BACH1 and mitochondrial metabolism. Nature 2019, 568, 254–258. [Google Scholar] [CrossRef]
- Frezza, C.; Zheng, L.; Folger, O.; Rajagopalan, K.N.; MacKenzie, E.D.; Jerby, L.; Micaroni, M.; Chaneton, B.; Adam, J.; Hedley, A.; et al. Haem oxygenase is synthetically lethal with the tumour suppressor fumarate hydratase. Nature 2011, 477, 225–228. [Google Scholar] [CrossRef]
- Han, L.; Jiang, J.; Ma, Q.; Wu, Z.; Wang, Z. The inhibition of heme oxygenase-1 enhances the chemosensitivity and suppresses the proliferation of pancreatic cancer cells through the SHH signaling pathway. Int. J. Oncol. 2018, 52, 2101–2109. [Google Scholar] [CrossRef]
- Miyake, M.; Fujimoto, K.; Anai, S.; Ohnishi, S.; Nakai, Y.; Inoue, T.; Matsumura, Y.; Tomioka, A.; Ikeda, T.; Okajima, E.; et al. Inhibition of heme oxygenase-1 enhances the cytotoxic effect of gemcitabine in urothelial cancer cells. Anticancer Res. 2010, 30, 2145–2152. [Google Scholar]
- Chen, C.P.; Chen, K.; Feng, Z.; Wen, X.; Sun, H. Synergistic antitumor activity of artesunate and HDAC inhibitors through elevating heme synthesis. Acta Pharm. Sin. B 2019, 9, 937–951. [Google Scholar] [CrossRef]
- Wang, J.; Zhang, J.; Shi, Y.; Xu, C.; Zhang, C.; Wong, Y.K.; Lee, Y.M.; Krishna, S.; He, Y.; Lim, T.K.; et al. Mechanistic Investigation of the Specific Anticancer Property of Artemisinin and Its Combination with Aminolevulinic Acid for Enhanced Anticolorectal Cancer Activity. ACS Cent. Sci. 2017, 3, 743–750. [Google Scholar] [CrossRef] [PubMed]
- Lin, K.H.; Xie, A.; Rutter, J.C.; Ahn, Y.R.; Lloyd-Cowden, J.M.; Nichols, A.G.; Soderquist, R.S.; Koves, T.R.; Muoio, D.M.; MacIver, N.J.; et al. Systematic Dissection of the Metabolic-Apoptotic Interface in AML Reveals Heme Biosynthesis to Be a Regulator of Drug Sensitivity. Cell Metab. 2019, 29, 1217–1231.e7. [Google Scholar] [CrossRef] [PubMed]
- Reita, D.; Bour, C.; Benbrika, R.; Groh, A.; Pencreach, E.; Guérin, E.; Guenot, D. Synergistic Anti-Tumor Effect of mTOR Inhibitors with Irinotecan on Colon Cancer Cells. Cancers 2019, 11, 1581. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Raffaele, M.; Pittalà, V.; Zingales, V.; Barbagallo, I.; Salerno, L.; Li Volti, G.; Romeo, G.; Carota, G.; Sorrenti, V.; Vanella, L. Heme Oxygenase-1 Inhibition Sensitizes Human Prostate Cancer Cells towards Glucose Deprivation and Metformin-Mediated Cell Death. Int. J. Mol. Sci. 2019, 20, 2593. [Google Scholar] [CrossRef] [Green Version]
- Biller, L.H.; Schrag, D. Diagnosis and Treatment of Metastatic Colorectal Cancer: A Review. JAMA 2021, 325, 669–685. [Google Scholar] [CrossRef] [PubMed]
- Sohn, K.J.; Croxford, R.; Yates, Z.; Lucock, M.; Kim, Y.I. Effect of the methylenetetrahydrofolate reductase C677T polymorphism on chemosensitivity of colon and breast cancer cells to 5-fluorouracil and methotrexate. J. Natl. Cancer Inst. 2004, 96, 134–144. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lu, J.; Hu, Y.; Qian, R.; Zhang, Y.; Yang, X.; Luo, P. Enhanced proliferation inhibition and apoptosis in glioma cells elicited by combination of irinotecan and imatinib. Eur. J. Pharmacol. 2020, 874, 173022. [Google Scholar] [CrossRef]
- Murono, K.; Tsuno, N.H.; Kawai, K.; Sasaki, K.; Hongo, K.; Kaneko, M.; Hiyoshi, M.; Tada, N.; Nirei, T.; Sunami, E.; et al. SN-38 overcomes chemoresistance of colorectal cancer cells induced by hypoxia, through HIF1alpha. Anticancer Res. 2012, 32, 865–872. [Google Scholar]
- Wibom, R.; Hagenfeldt, L.; von Döbeln, U. Measurement of ATP production and respiratory chain enzyme activities in mitochondria isolated from small muscle biopsy samples. Anal. Biochem. 2002, 311, 139–151. [Google Scholar] [CrossRef]
- Zhu, X.G.; Chudnovskiy, A.; Baudrier, L.; Prizer, B.; Liu, Y.; Ostendorf, B.N.; Yamaguchi, N.; Arab, A.; Tavora, B.; Timson, R.; et al. Functional Genomics In Vivo Reveal Metabolic Dependencies of Pancreatic Cancer Cells. Cell Metab. 2021, 33, 211–221.e6. [Google Scholar] [CrossRef] [PubMed]
- Lignitto, L.; LeBoeuf, S.E.; Homer, H.; Jiang, S.; Askenazi, M.; Karakousi, T.R.; Pass, H.I.; Bhutkar, A.J.; Tsirigos, A.; Ueberheide, B.; et al. Nrf2 Activation Promotes Lung Cancer Metastasis by Inhibiting the Degradation of Bach1. Cell 2019, 178, 316–329.e18. [Google Scholar] [CrossRef] [PubMed]
- Wiel, C.; Le Gal, K.; Ibrahim, M.X.; Jahangir, C.A.; Kashif, M.; Yao, H.; Ziegler, D.V.; Xu, X.; Ghosh, T.; Mondal, T.; et al. BACH1 Stabilization by Antioxidants Stimulates Lung Cancer Metastasis. Cell 2019, 178, 330–345.e22. [Google Scholar] [CrossRef]
- Memmott, R.M.; Mercado, J.R.; Maier, C.R.; Kawabata, S.; Fox, S.D.; Dennis, P.A. Metformin prevents tobacco carcinogen--induced lung tumorigenesis. Cancer Prev. Res. 2010, 3, 1066–1076. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mohammed, A.; Janakiram, N.B.; Brewer, M.; Ritchie, R.L.; Marya, A.; Lightfoot, S.; Steele, V.E.; Rao, C.V. Antidiabetic Drug Metformin Prevents Progression of Pancreatic Cancer by Targeting in Part Cancer Stem Cells and mTOR Signaling. Transl. Oncol. 2013, 6, 649–659. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hou, M.; Venier, N.; Sugar, L.; Musquera, M.; Pollak, M.; Kiss, A.; Fleshner, N.; Klotz, L.; Venkateswaran, V. Protective effect of metformin in CD1 mice placed on a high carbohydrate-high fat diet. Biochem. Biophys. Res. Commun. 2010, 397, 537–542. [Google Scholar] [CrossRef]
- Schneider, M.B.; Matsuzaki, H.; Haorah, J.; Ulrich, A.; Standop, J.; Ding, X.Z.; Adrian, T.E.; Pour, P.M. Prevention of pancreatic cancer induction in hamsters by metformin. Gastroenterology 2001, 120, 1263–1270. [Google Scholar] [CrossRef]
- Zhao, B.; Luo, J.; Yu, T.; Zhou, L.; Lv, H.; Shang, P. Anticancer mechanisms of metformin: A review of the current evidence. Life Sci. 2020, 254, 117717. [Google Scholar] [CrossRef]
- Foretz, M.; Guigas, B.; Bertrand, L.; Pollak, M.; Viollet, B. Metformin: From mechanisms of action to therapies. Cell Metab. 2014, 20, 953–966. [Google Scholar] [CrossRef] [Green Version]
- Owen, M.R.; Doran, E.; Halestrap, A.P. Evidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondrial respiratory chain. Biochem. J. 2000, 348 Pt 3, 607–614. [Google Scholar] [CrossRef]
- Anisimov, V.N. Metformin for cancer and aging prevention: Is it a time to make the long story short? Oncotarget 2015, 6, 39398–39407. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hosono, K.; Endo, H.; Takahashi, H.; Sugiyama, M.; Sakai, E.; Uchiyama, T.; Suzuki, K.; Iida, H.; Sakamoto, Y.; Yoneda, K.; et al. Metformin suppresses colorectal aberrant crypt foci in a short-term clinical trial. Cancer Prev. Res. 2010, 3, 1077–1083. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tsai, M.J.; Yang, C.J.; Kung, Y.T.; Sheu, C.C.; Shen, Y.T.; Chang, P.Y.; Huang, M.S.; Chiu, H.C. Metformin decreases lung cancer risk in diabetic patients in a dose-dependent manner. Lung Cancer 2014, 86, 137–143. [Google Scholar] [CrossRef] [PubMed]
- Lin, J.J.; Gallagher, E.J.; Sigel, K.; Mhango, G.; Galsky, M.D.; Smith, C.B.; LeRoith, D.; Wisnivesky, J.P. Survival of patients with stage IV lung cancer with diabetes treated with metformin. Am. J. Respir. Crit. Care Med. 2015, 191, 448–454. [Google Scholar] [CrossRef] [Green Version]
- Currie, C.J.; Poole, C.D.; Gale, E.A. The influence of glucose-lowering therapies on cancer risk in type 2 diabetes. Diabetologia 2009, 52, 1766–1777. [Google Scholar] [CrossRef] [Green Version]
- Li, D.; Yeung, S.C.; Hassan, M.M.; Konopleva, M.; Abbruzzese, J.L. Antidiabetic therapies affect risk of pancreatic cancer. Gastroenterology 2009, 137, 482–488. [Google Scholar] [CrossRef] [Green Version]
- Xin, W.; Fang, L.; Fang, Q.; Zheng, X.; Huang, P. Effects of metformin on survival outcomes of pancreatic cancer patients with diabetes: A meta-analysis. Mol. Clin. Oncol. 2018, 8, 483–488. [Google Scholar] [CrossRef]
- Shi, Y.Q.; Zhou, X.C.; Du, P.; Yin, M.Y.; Xu, L.; Chen, W.J.; Xu, C.F. Relationships are between metformin use and survival in pancreatic cancer patients concurrent with diabetes: A systematic review and meta-analysis. Medicine 2020, 99, e21687. [Google Scholar] [CrossRef]
- Lee, J.H.; Kim, T.I.; Jeon, S.M.; Hong, S.P.; Cheon, J.H.; Kim, W.H. The effects of metformin on the survival of colorectal cancer patients with diabetes mellitus. Int. J. Cancer 2012, 131, 752–759. [Google Scholar] [CrossRef]
- Rajgopal, R.K.; Kochhar, R.S. Efficacy and Cardiovascular Safety of Metformin. Curr. Drug Saf. 2021, 16, 165–177. [Google Scholar] [CrossRef]
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Allocco, A.L.; Bertino, F.; Petrillo, S.; Chiabrando, D.; Riganti, C.; Bardelli, A.; Altruda, F.; Fiorito, V.; Tolosano, E. Inhibition of Heme Export and/or Heme Synthesis Potentiates Metformin Anti-Proliferative Effect on Cancer Cell Lines. Cancers 2022, 14, 1230. https://doi.org/10.3390/cancers14051230
Allocco AL, Bertino F, Petrillo S, Chiabrando D, Riganti C, Bardelli A, Altruda F, Fiorito V, Tolosano E. Inhibition of Heme Export and/or Heme Synthesis Potentiates Metformin Anti-Proliferative Effect on Cancer Cell Lines. Cancers. 2022; 14(5):1230. https://doi.org/10.3390/cancers14051230
Chicago/Turabian StyleAllocco, Anna Lucia, Francesca Bertino, Sara Petrillo, Deborah Chiabrando, Chiara Riganti, Alberto Bardelli, Fiorella Altruda, Veronica Fiorito, and Emanuela Tolosano. 2022. "Inhibition of Heme Export and/or Heme Synthesis Potentiates Metformin Anti-Proliferative Effect on Cancer Cell Lines" Cancers 14, no. 5: 1230. https://doi.org/10.3390/cancers14051230