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01-08-2008 | Preclinical Study

Alteration of expression levels of the oxidative phosphorylation system (OXPHOS) in breast cancer cell mitochondria

Authors: Lorenza Putignani, Salvatore Raffa, Roberta Pescosolido, Laura Aimati, Fabrizio Signore, Maria Rosaria Torrisi, Paola Grammatico

Published in: Breast Cancer Research and Treatment | Issue 3/2008

Abstract

Mitochondria are dynamic intracellular organelles playing a central role in cell metabolism by generating ATP, through the oxidative phosphorylation system (OXPHOS). Altered mitochondrial functions have been identified as causative or contributing factors in some degenerative diseases and are becoming crucial to understanding cancer mechanisms. We report on distinct expression differences between mitochondria of normal and breast-infiltrating ductal carcinoma (IDC) cells. Mitochondria isolated from HMC (human mammary carcinoma) and HMEC (human mammary epithelial cell) cultures were assayed for expression levels of the multi-protein OXPHOS complexes using Western blot and densitometric analyses. Depressed expression levels were detected for all HMC OXPHOS complexes. Drastic signal reduction was observed for the succinate-dehydrogenase complex II iron-sulphur protein SDH-B (3.38%), while decreasing was reported for the NADH-ubiquinone oxidoreductase complex I Fe-S protein 3 NDUFS3 (32.78%) and the ubiquinol-cytochrome c reductase complex III protein 2 UQCRC2 (50.34%). A significant signal dropping was detected for the ATP-synthase complex V F₁β subunit (18.07%). For the cytochrome-oxidase complex IV (CO), near-depletion of the mitochondrial-encoded COI (4.37%) and no apparent variation of the COIV (97.26%) subunits were observed. CO and ATP-synthase were also assayed by cryo-immunoelectron microscopy (CIEM) on unfractionated HMC and HEMC cell mitochondria. COI and F₁β differential expression, invariance of COIV levels were corroborated, while HMC mitochondria morphology deterioration was highlighted. MitoTracker Red and fluorescence immunolabelling merging confirmed CIEM data. MitoTracker Red and Green co-staining showed mitochondria membrane property modulation. These data describe bioenergetic and phenotypic alterations of IDC cell mitochondria, possibly providing new cancer hallmarks.

Warburg O (1929) Ist die aerobe Glykolyse spezifish fur die Tumoren? Biochem Z 204:482–483

Warburg O (1956) On the origin of cancer cells. Science 123:309–314PubMedCrossRef

Levonen AL, Patel RP, Brookes P et al (2001) Mechanisms of cell signaling by nitric oxide and peroxynitrite: from mitochondria to MAP kinases. Antioxid Redox Signal 3:215–229PubMedCrossRef

Bogoyevitch MA, Ng DC, Court NW (2000) Intact mitochondrial electron transport function is essential for signalling by hydrogen peroxide in cardiac myocytes. J Mol Cell Cardiol 32:1469–1480PubMedCrossRef

Droge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95PubMed

Liu X, Kim CN, Yang J et al (1996) Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 86:147–157PubMedCrossRef

Electronic biology and cancer (1997) In: Kaminer B (ed) Search and discovery: a tribute to albert szent-gyorgyi. Academic Press, New York, pp 329–335

Woods MW, DuBuy HG (1945) Cytoplasmic diseases and cancer. Science 102:591–593PubMedCrossRef

Singh KK (1998) Mitochondrial DNA mutations in aging, disease, and cancer. Springer, New York

10.

Polyak K, Li Y, Zhu H et al (1998) Somatic mutations of the mitochondrial genome in human colorectal tumours. Nat Genet 20:291–293PubMedCrossRef

11.

Kroemer G, Reed JC (2000) Mitochondrial control of cell death. Nat Med 6:513–519PubMedCrossRef

12.

Brandon M, Baldi P, Wallace DC (2006) Mitochondrial mutations in cancer. Oncogene 25:4647–4662PubMedCrossRef

13.

Isidoro A, Martinez M, Fernandez PL et al (2004) Alteration of the bioenergetic phenotype of mitochondria is a hallmark of breast, gastric, lung and oesophageal cancer. Biochem J 378:17–20PubMedCrossRef

14.

Isidoro A, Casado E, Redondo A et al (2005) Breast carcinomas fulfill the Warburg hypothesis and provide metabolic markers of cancer prognosis. Carcinogenesis 26:2095–2104 PubMedCrossRef

15.

Scheffler IE (1999) Mitochondria. Wiley-Liss, New York

16.

Benit P, Slama A, Cartault F et al (2004) Mutant NDUFS3 subunit of mitochondrial complex I causes Leigh syndrome. J Med Genet 41:14–17PubMedCrossRef

17.

Ugalde C, Vogel R, Huijbens R et al (2004) Human mitochondrial complex I assembles through the combination of evolutionary conserved modules: a framework to interpret complex I deficiencies. Human Mol Genet 20:2461–2472CrossRef

18.

Au HC, Ream-Robinson D, Bellew LA et al (1995) Structural organization of the gene encoding the human iron-sulfur subunit of succinate dehydrogenase. Gene 159:249–253PubMedCrossRef

19.

Ackrell BA (2000) Progress in understanding structure-function relationships in respiratory chain complex II. FEBS Lett. 466:1–5PubMedCrossRef

20.

Duncan AM, Ozawa T, Suzuki H et al (1993) Assignment of the gene for the core protein II (UQCRC2) subunit of the mitochondrial cytochrome bc1 complex to human chromosome 16p12. Genomics 18:455–456PubMedCrossRef

21.

Coenen MJ, van den Heuvel LP, Smeitink JA (2001) Mitochondrial oxidative phosphorylation system assembly in man: recent achievements. Curr Opin Neurol 14:777–781PubMedCrossRef

22.

Tsukihara T, Aoyama H, Yamashita E et al (1996) The whole structure of the 13-subunit oxidized cytochrome c oxidase at 2.8 A. Science 272:1136–1144PubMedCrossRef

23.

Groen AK, Wanders RJ, Westerhoff HV et al (1982) Quantification of the contribution of various steps to the control of mitochondrial respiration. J Biol Chem 257:2754–2757PubMed

24.

Higuti T, Tsurumi C, Osaka F et al (1991) Molecular cloning of cDNA for the import precursor of human subunit β of H(+)-ATP synthase in mitochondria. Biochem Biophys Res Commun 178:1014–1020PubMedCrossRef

25.

Schon EA, Santra S, Pallotti F et al (2001) Pathogenesis of primary defects in mitochondrial ATP synthesis. Semin Cell Dev Biol 12:441–448 PubMedCrossRef

26.

Villani G, Attardi G (2000) In vivo control of respiration by cytochrome c oxidase in human cells. Free Radic Biol Med 29:202–210PubMedCrossRef

27.

Vijayasarathy C, Biunno I, Lenka Net et al (1998) Variations in the subunit content and catalytic activity of the cytochrome c oxidase complex from different tissues and different cardiac compartments. Biochim Biophys Acta 1371:71–82PubMedCrossRef

28.

Speirs V, Green AR, Walton DS et al (1998) Short-term primary culture of epithelial cells derived from human breast tumours. Br J Cancer 78:1421–1429PubMed

29.

Emerman JT, Wilkinson DA (1990) Routine culturing of normal, dysplastic and malignant human mammary epithelial cells from small tissue samples. In Vitro Cell Dev Biol 26:1186–1194PubMedCrossRef

30.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685CrossRef

31.

Barber RD, Harmer DW, Coleman RA et al (2005) GADPH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics 21:389–395 PubMedCrossRef

32.

Herrmann PC, Gillespie JW, Charboneau L et al (2003) Mitochondrial proteome: altered cytochrome c oxidase subunit levels in prostate cancer. Proteomics 3:1801–1810PubMedCrossRef

33.

Krieg RC, Knuechel R, Schiffmann E et al (2004) Mitochondrial proteome: cancer-altered metabolism associated with cytochrome c oxidase subunit level variation. Proteomics 4:2789–2795PubMedCrossRef

34.

Cuezva JM, Chen G, Alonso AM et al (2004) The bioeneergetic signature of lung adenocarcinomas is a molecular marker of cancer diagnosis and prognosis. Carcinogenesis 25:1157–1163PubMedCrossRef

35.

Lemasters JJ (2007) Modulation of mitochondrial membrane permeability in pathogenesis, authophagy and control of metabolism. J Gastroenterology Hepatology 22:S31–S37CrossRef

36.

Rustin P, Rötig A (2002) Inborn errors of complex II-unusual human mitochondrial diseases. Biochim Biophys Acta 1553:117–122PubMedCrossRef

37.

Chen Q, Vazquez EJ, Moghaddas S et al (2003) Production of reactive oxygen species by mitochondria: central role of complex III. J Biol Chem 278:36027–36031PubMedCrossRef

38.

Felty Q, Roy D (2005) Estrogen, mitochondria, and growth of cancer and non-cancer cells. J Carcinog 4:1–18PubMedCrossRef

39.

Chen JQ, Yager JD, Russo J (2005) Regulation of mitochondrial respiratory chain structure and function by estrogens/estrogen receptors and potential physiological/pathophysiological implications. Biochim Biophys Acta 1746:1–17PubMedCrossRef

40.

Felty Q, Singh KP, Roy D (2005) Estrogen-induced G1/S transition of G0-arrested estrogen-dependent breast cancer cells is regulated by mitochondrial oxidant signaling. Oncogene 24:4883–4893PubMedCrossRef

41.

Felty Q, Xiong WC, Sun D et al (2005) Estrogen-induced mitochondrial reactive oxygen species as signal-transducing messengers. Biochemistry 44:6900–6909PubMedCrossRef

42.

Gibson BW (2005) The human mitochondrial proteome: oxidative stress, protein modifications and oxidative phosphorylation. Int J Biochem Cell Biol 37:927–934PubMedCrossRef

43.

Patwardhan AJ, Strittmatter EF, Camp DG 2nd et al (2005) Comparison of normal and breast cancer cell lines using proteome, genome, and interactome data. J Proteome Res 4:1952–1960PubMedCrossRef

44.

Moreira PI, Custodio J, Moreno A et al (2006) Tamoxifen and estradiol interact with the flavin mononucleotide site of complex I leading to mitochondrial failure. J Biol Chem 281:10143–10152PubMedCrossRef

45.

Knowles HJ, Harris AL (2001) Hypoxia and oxidative stress in breast cancer. Hypoxia and tumourigenesis. Breast Cancer Res 3:318–322CrossRef

46.

Izquierdo JM, Cuezva JM (2005) Epigenetic regulation of the binding activity of translation inhibitory proteins that bind the 3′ untranslated region of beta-F1-ATPase mRNA by adenine nucleotides and the redox state. Arch Biochem Biophys 433:481–486PubMedCrossRef

47.

Izquierdo JM, Cuezva JM (1997) Control of the translational efficiency of beta-F1-ATPase mRNA depends on the regulation of a protein that bind the 3′ untranslated region of the mRNA. Mol Cell Biol 17:5255–5268PubMed

48.

Schulz TJ, Thierbach R, Voigt A et al (2006) Induction of oxidative metabolism by mitochondrial frataxin inhibits cancer growth: Otto Warburg revisited. J Biol Chem 281:977–981PubMedCrossRef

49.

Modica-Napolitano JS, Singh K (2002) Mitochondria as targets for detection and treatment of cancer. Expert Rev Mol Med 11:1–19CrossRef

Title: Alteration of expression levels of the oxidative phosphorylation system (OXPHOS) in breast cancer cell mitochondria
Authors: Lorenza Putignani
Salvatore Raffa
Roberta Pescosolido
Laura Aimati
Fabrizio Signore
Maria Rosaria Torrisi
Paola Grammatico
Publication date: 01-08-2008
Publisher: Springer US
Published in: Breast Cancer Research and Treatment / Issue 3/2008
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-007-9738-x

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