PL EN


Preferences help
enabled [disable] Abstract
Number of results
2005 | 52 | 4 | 881-888
Article title

Expression and hypoxia-responsiveness of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 in mammary gland malignant cell lines

Content
Title variants
Languages of publication
EN
Abstracts
EN
Recently, we have shown that PFKFB4 gene which encodes the testis isoenzyme of PFKFB is also expressed in the prostate and hepatoma cancer cell lines. Here we have studied expression and hypoxic regulation of the testis isoenzyme of PFKFB4 in several malignant cell lines from a female organ - the mammary gland. Our studies clearly demonstrated that PFKFB4 mRNA is also expressed in mammary gland malignant cells (MCF-7 and T47D cell lines) in normoxic conditions and that hypoxia strongly induces it expression. To better understand the mechanism of hypoxic regulation of PFKFB4 gene expression, we used dimethyloxalylglycine, a specific inhibitor of HIF-1α hydroxylase enzymes, which strongly increases HIF-1α levels and mimics the effect of hypoxia. It was observed that PFKFB4 expression in the MCF7 and T47D cell lines was highly responsive to dimethyloxalylglycine, suggesting that the hypoxia responsiveness of PFKFB4 gene in these cell lines is regulated by HIF-1 proteins. Moreover, desferrioxamine and cobalt chloride, which mimic the effect of hypoxia by chelating or substituting for iron, had a similar stimulatory effect on the expression of PFKFB mRNA. In other mammary gland malignant cell lines (BT549, MDA-MB-468, and SKBR-3) hypoxia and hypoxia mimics also induced PFKFB4 mRNA, but to variable degrees. The hypoxic induction of PFKFB4 mRNA was equivalent to the expression of PFKFB3, Glut1, and VEGF, which are known HIF-1-dependent genes. Hypoxia and dimethyloxalylglycine increased the PFKFB4 protein levels in all cell lines studied except MDA-MB-468. Through site-specific mutagenesis in the 5'-flanking region of PFKFB4 gene the hypoxia response could be limited. Thus, this study provides evidence that PFKFB4 gene is also expressed in mammary gland cancer cells and strongly responds to hypoxia via an HIF-1α dependent mechanism. Moreover, the PFKFB4 and PFKFB3 gene expression in mammary gland cancer cells has also a significant role in the Warburg effect which is found in all malignant cells.
Year
Volume
52
Issue
4
Pages
881-888
Physical description
Dates
published
2005
received
2004-11-30
revised
2005-05-13
accepted
2005-06-05
(unknown)
2005-07-11
References
  • Armstead VE, Opentanova IL, Minchenko AG, Lefer AM (1999) Tissue factor gene expression in vital organs during murine traumatic shock: role of transcription factors AP-1 and NF-κB. Anesthesiology 91: 1844-1852.
  • Atsumi T, Chesney J, Metz C, Leng L, Donnelly S, Makita Z, Mitchell R, Bucala R (2002) High expression of inducible 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (iPFK-2; PFKFB3) in human cancers. Cancer Res 62: 5881-5887.
  • Brown JM, Giaccia AM (1998) The unique physiology of solid tumors: opportunities (and problems) for cancer therapy. Cancer Res 58: 1408-1416.
  • Chesney J, Mitchell R, Benigni F, Bacher M, Spiegel L, Al-Abed Y, Han JH, Metz C, Bucala R (1999) An inducible gene product for 6-phosphofructo-2-kinase with an AU-rich instability element: role in tumor cell glycolysis and the Warburg effect. Proc Natl Acad Sci USA 96: 3047-3052.
  • Crepin KM, Darville MI, Hue L, Rousseau GG (1989) Characterization of distinct mRNAs coding for putative isozymes of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Eur J Biochem 183: 433-440.
  • Dang CV, Semenza GL (1999) Oncogenic alterations of metabolism. Trends Biochem Sci 24: 68-76.
  • Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O'Rourke J, Mole DR, Mukherji M, Metzen E, Wilson MI, Dhanda A, Tian YM, Masson N, Hamilton DL, Jaakkola P, Barstead R, Hodgkin J, Maxwell PH, Pugh CW, Schofield CJ, Ratcliffe PJ (2001) C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell 107: 43-54.
  • Gleade JM, Ratcliffe PJ (1998) Hypoxia and the regulation of gene expression. Mol Med Today 4: 122-129.
  • Gleadle JM, Ebert BL, Firth JD, Ratcliffe PJ (1995) Regulation of angiogenic growth factor expression by hypoxia, transition metals, and chelating agents. Am J Physiol 268: C1362-C1368.
  • Hockel M, Vaupel P (2001) Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects. J Natl Cancer Inst 93: 266-276.
  • Hopfl G, Ogunshola O, Gassmann M (2004) HIFs and tumors - causes and consequences. Am J Physiol Regul Integr Comp Physiol 286: R608-R623.
  • Hue L, Beauloye C, Bertrand L, Horman S, Krause U, Marsin A-S, Meisse D, Vertommen D, Rider MH (2003) New targets of AMP-activated protein kinase. Biochem Soc Trans 31: 213-215.
  • Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M, Salic A, Asara JM, Lane WS, Kaelin WGJ (2001) HIFα targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 292: 464-468.
  • Lando D, Peet DJ, Whelan DA, Gorman JJ, Whitelaw ML (2002) Asparagine hydroxylation of the HIF transactivation domain: a hypoxic switch. Science 295: 858-861.
  • Lu H, Forbes RA, Verma A (2002) Hypoxia-inducible factor 1 activation by aerobic glycolysis implicates the Warburg effect in carcinogenesis. J Biol Chem 277: 23111-23115.
  • Manzano A, Rosa JL, Ventura F, Perez JX, Nadal M, Estivill X, Ambrosio S, Gil J, Bartrons R (1998) Cloning, expression and chromosomal localization of a human testis 6-phosphofructo-2-kinase/fructose-2,6-bisphospha­tase gene. Gene 229: 83-89.
  • Marsin AS, Douzin C, Bertrand L, Hue L (2002) The stimulation of glycolysis by hypoxia in activated monocytes is mediated by AMP-activated protein kinase and inducible 6-phosphofructo-2-kinase. J Biol Chem 277: 30778-30783.
  • Masson N, Ratcliffe PJ (2003) HIF prolyl and asparaginyl hydroxylases in the biological response to intracellular O2 levels. J Cell Sci 116: 3041-3049.
  • Metzen E, Ratcliffe PJ (2004) HIF hydroxylation and cellular oxygen sensing. Biol Chem 385: 223-230.
  • Min J-H, Yang H, Ivan M, Gertler F, Kaelin WGJ Jr, Pavletich NP (2002) Structure of an HIF-1α-pVHL complex: hydroxyproline recognition in signaling. Science 296: 1886-1889.
  • Minchenko A, Caro J (2000) Regulation of endothelin-1 gene expression in human microvascular endothelial cells by hypoxia and cobalt: role of hypoxia responsive element. Mol Cell Biochem 208: 53-62.
  • Minchenko A, Bauer T, Salceda S, Caro J (1994) Hypoxic stimulation of vascular endothelial growth factor expression in vitro and in vivo. Lab Invest 71: 374-379.
  • Minchenko A, Leshchinsky I, Opentanova I, Sang N, Srinivas V, Armstead V, Caro J (2002) Hypoxia-inducible factor-1-mediated expression of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) gene: its possible role in the Warburg effect. J Biol Chem 277: 6183-6187.
  • Minchenko O, Opentanova I, Caro J (2003) Hypoxic regulation of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene family (PFKFB-1-4) expression in vivo. FEBS Lett 554: 264-270.
  • Minchenko OH, Opentanova IL, Minchenko DO, Ogura T, Esumi H (2004) Hypoxia induces transcription of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 gene via hypoxia inducible factor-1alpha activation. FEBS Lett 576: 14-20.
  • Mole DR, Schlemminger I, McNeil LA, Hewitson KS, Pugh CW, Ratcliffe PJ, Schofield CJ (2003) 2-Oxoglutarate analogue inhibitors of HIF prolyl hydroxylase. Bioorg Med Chem Lett 13, 2677-2680.
  • Mueller MD, Vigne J-L, Minchenko A, Lebovic DI, Leitman DC, Taylor RN (2000) Regulation of vascular endothelial growth factor (VEGF) gene transcription by estrogen receptors α and β. Proc Natl Acad Sci USA 97: 10972-10977.
  • Okar DA, Manzano A, Navarro-Sabate A, Riera L, Bartrons R, Lange A (2001) PFK-2/FBPase-2: maker and breaker of the essential biofactor fructose-2,6-bisphosphate. Trends Biochem Sci 26: 30-35.
  • Pilkis SJ, Claus TH, Kurland IJ, Lange AJ (1995) 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase: a metabolic signaling enzyme. Annu Rev Biochem 64: 799-835.
  • Rider MH, Bertrand L, Vertommen D, Michels PA, Rousseau GG, Hue L (2004) 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase: head-head with a bifunctional enzyme that controls glycolysis. Biochem J 381: 561-578.
  • Sakakibara R, Okudaira T, Fujiwara K, Kato M, Hirata T, Yamanaka S, Naito M, Fukasawa M (1999) Tissue distribution of placenta-type 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Biochem Biophys Res Commun 257: 177-181.
  • Sakata J, Abe Y, Uyeda K (1991) Molecular cloning of the DNA and expression and characterization of rat testes fructose-6-phosphate,2-kinase:fructose-2,6-bisphosphatase. J Biol Chem 266: 15764-15770.
  • Schofield CJ, Ratcliffe PJ (2004) Oxygen sensing by HIF hydroxylases. Nat Rev Mol Cell Biol 5: 343-354.
  • Vaupel P (1996) Oxygen transport in tumors: characteristics and clinical implications. Adv Exp Med Biol 388: 341-351.
  • Warburg O, Wind F, Negalein E (1927) The metabolism of tumours in the body. J Physiol 8: 519-530.
  • Warburg O (1956) On respiratory impairment in cancer cells. Science 123: 309-314.
  • Wenger RH (2002) Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression. FASEB J 16: 1151-1162.
Document Type
Publication order reference
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv52p881kz
Identifiers
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.