Although the Warburg effect is a dominant metabolic phenotype observed in cancers, the metabolic changes and adaptation occurring in tumors have been demonstrated to extend beyond the Warburg effect and thus considered a secondary effect to the transformation process of carcinogenesis, including nutritional deficiencies. However, the role of nutritional deficiencies in this metabolic reprogramming (e. g., oxidative phosphorylation (OXPHOS)/glycolysis interconversion) is not completely known yet. Here, we showed that under regular culture condition, the proliferation of U251 cells, but not other tumor cell lines, preferentially performed the Warburg effect and was remarkably inhibited by oxamic acid which can inhibit the activity of lactate dehydrogenase (LDH); whereas under serum starvation, glycolysis was depressed, tricarboxylic acid cycle (TCA) was enhanced, and the activity of OXPHOS was reinforced to maintain cellular ATP content in a high level, but interestingly, we observed a decreased expression of reactive oxygen species (ROS). Moreover, the upregulated activity of mitochondrial complex I was confirmed by Western blots and showed that the mitochondrial-related protein, NDUFA9, NDUFB8, ND1, and VDAC1 were remarkably increased after serum starved. Mechanistically, nutritional deficiencies could reduce hypoxia-inducible factor α (HIF-1α) protein expression to increase C-MYC protein level, which in turn increased nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM) transcription to enhance the activity of OXPHOS, suggesting that metabolic reprogramming by the changes of microenvironment during the carcinogenesis can provide some novel therapeutic clues to traditional cancer treatments.
Starvation OXPHOS Glycolysis HIF-1α C-MYC
This is a preview of subscription content, log in to check access.
We would like to thank all the members of our laboratory for the encouragement and help in this study. This work was financially supported by the National Natural Science Foundation of China (no. 31260276, no. 31160237, and no. 81271330), the talent program of Yunnan Province (no. W8110305), and the Yunnan Province Science and Technology Innovation Team (no. 2011CI123)
ZJL, YS, and MY designed the experiments with valuable help from QHC, MZL, HUH and ZJL performed and analyzed data with valuable help from ST and LL, and ZJL wrote the manuscript. ZJL and MY oversaw the overall project.
Compliance with ethical standards
Conflicts of interest
Moreno-Sanchez R, Marin-Hernandez A, Saavedra E, Pardo JP, Ralph SJ, Rodriguez-Enriquez S. Who controls the ATP supply in cancer cells? Biochemistry lessons to understand cancer energy metabolism. Int J Biochem Cell Biol. 2014;50:10–23. doi:10.1016/j.biocel.2014.01.025.CrossRefPubMedGoogle Scholar
Bustamante E, Morris HP, Pedersen PL. Energy metabolism of tumor cells. Requirement for a form of hexokinase with a propensity for mitochondrial binding. J Biol Chem. 1981;256(16):8699–704.PubMedGoogle Scholar
Bonuccelli G, Tsirigos A, Whitaker-Menezes D, Pavlides S, Pestell RG, Chiavarina B, et al. Ketones and lactate “fuel” tumor growth and metastasis: evidence that epithelial cancer cells use oxidative mitochondrial metabolism. Cell Cycle. 2010;9(17):3506–14.CrossRefPubMedPubMedCentralGoogle Scholar
Bashan N, Burdett E, Hundal HS, Klip A. Regulation of glucose transport and GLUT1 glucose transporter expression by O2 in muscle cells in culture. Am J Physiol. 1992;262(3 Pt 1):C682–90.PubMedGoogle Scholar
Signorile A, Micelli L, De Rasmo D, Santeramo A, Papa F, Ficarella R, et al. Regulation of the biogenesis of OXPHOS complexes in cell transition from replicating to quiescent state: involvement of PKA and effect of hydroxytyrosol. Biochim Biophys Acta. 2014;1843(4):675–84. doi:10.1016/j.bbamcr.2013.12.017.CrossRefPubMedGoogle Scholar
Yu M, Dai J, Huang W, Jiao Y, Liu L, Wu M, et al. hMTERF4 knockdown in HeLa cells results in sub-G1 cell accumulation and cell death. Acta Biochim Biophys Sin (Shanghai). 2011;43(5):372–9. doi:10.1093/abbs/gmr020.CrossRefGoogle Scholar
Zhang E, Li X, Zhang S, Chen L, Zheng X. Cell cycle synchronization of embryonic stem cells: effect of serum deprivation on the differentiation of embryonic bodies in vitro. Biochem Biophys Res Commun. 2005;333(4):1171–7. doi:10.1016/j.bbrc.2005.05.200.CrossRefPubMedGoogle Scholar
Hsiao YP, Lai WW, Wu SB, Tsai CH, Tang SC, Chung JG, et al. Triggering apoptotic death of human epidermal keratinocytes by malic acid: involvement of endoplasmic reticulum stress- and mitochondria-dependent signaling pathways. Toxins (Basel). 2015;7(1):81–96. doi:10.3390/toxins7010081.CrossRefGoogle Scholar
Nazmara Z, Salehnia M, HosseinKhani S. Mitochondrial distribution and ATP content of vitrified, in vitro matured mouse oocytes. Avicenna J Med Biotechnol. 2014;6(4):210–7.PubMedPubMedCentralGoogle Scholar
Xiong W, Huang W, Jiao Y, Ma J, Yu M, Ma M, et al. Production, purification and characterization of mouse monoclonal antibodies against human mitochondrial transcription termination factor 2 (MTERF2). Protein Expr Purif. 2012;82(1):11–9. doi:10.1016/j.pep.2011.10.012.CrossRefPubMedGoogle Scholar
Ratcliffe PJ. From erythropoietin to oxygen: hypoxia-inducible factor hydroxylases and the hypoxia signal pathway. Blood Purif. 2002;20(5):445–50.CrossRefPubMedGoogle Scholar
Semenza GL, Rue EA, Iyer NV, Pang MG, Kearns WG. Assignment of the hypoxia-inducible factor 1alpha gene to a region of conserved synteny on mouse chromosome 12 and human chromosome 14q. Genomics. 1996;34(3):437–9. doi:10.1006/geno.1996.0311.CrossRefPubMedGoogle Scholar
He M, Wang QY, Yin QQ, Tang J, Lu Y, Zhou CX, et al. HIF-1alpha downregulates miR-17/20a directly targeting p21 and STAT3: a role in myeloid leukemic cell differentiation. Cell Death Differ. 2013;20(3):408–18. doi:10.1038/cdd.2012.130.CrossRefPubMedGoogle Scholar
Zhang H, Gao P, Fukuda R, Kumar G, Krishnamachary B, Zeller KI, et al. HIF-1 inhibits mitochondrial biogenesis and cellular respiration in VHL-deficient renal cell carcinoma by repression of C-MYC activity. Cancer Cell. 2007;11(5):407–20. doi:10.1016/j.ccr.2007.04.001.CrossRefPubMedGoogle Scholar
Chen G, Dai J, Tan S, Meng S, Liu Z, Li M, et al. MTERF1 regulates the oxidative phosphorylation activity and cell proliferation in HeLa cells. Acta Biochim Biophys Sin (Shanghai). 2014;46(6):512–21. doi:10.1093/abbs/gmu029.CrossRefGoogle Scholar
Wu H, Ding Z, Hu D, Sun F, Dai C, Xie J, et al. Central role of lactic acidosis in cancer cell resistance to glucose deprivation-induced cell death. J Pathol. 2012;227(2):189–99. doi:10.1002/path.3978.CrossRefPubMedGoogle Scholar