Skip to main content


We’d like to understand how you use our websites in order to improve them. Register your interest.

The transcriptional cascade associated with creatine kinase down-regulation and mitochondrial biogenesis in mice sarcoma


The tissue-specific expressions of creatine kinase (CK) isoforms are regulated by the coordinated action of various transcription factors. The myogenic differentiation factor D (MyoD) family of proteins and the myocyte-specific enhancer binding factor 2 family of transcription factors are important in regulating the muscle-specific expression of cytosolic muscle-type CK (MCK) and mitochondrial CKs. As reported in some related studies, TNF-α mediated degradation of MyoD and myogenin mRNA may lead to severe muscle wasting and cachexia, which is characterized by a low transcript level of MCK and myosin heavy chain proteins. In our previous study, we reported on a complete loss of total CK activity and expression when sarcoma was induced in mouse skeletal muscle (Patra et al. FEBS J. 275 (2008) 3236–3247). This study aimed at investigating the transcriptional cascade of CK down-regulation in carcinogen-induced sarcoma in mouse muscle. Both CK deficiency and enhanced nitric oxide synthase (NOS) were known to augment mitochondrial biogenesis, so we also explored the activation of the transcriptional cascade of mitochondrial biogenesis in this cancer. We observed the activation of the TNF-α-mediated nitric oxide production pathway with NFκB activation and concomitant degradation of MyoD and myogenin mRNA. Exploration of mitochondrial biogenesis revealed high cytochrome c oxidase activity and mitochondrial DNA content in sarcoma. The PGC-related co-activator seems to have a major role in regulating mitochondrial biogenesis by upregulating nuclear respiratory factors and mitochondrial transcription factor A. From the above findings, it can be concluded that severe muscle degeneration leads to CK down-regulation in sarcoma, and that the stimulation of mitochondrial biogenesis indicated a scenario representing both CK deficiency and NOS overexpression on the one hand, and altered bioenergetic profiling on the other.



creatine kinase


cytochrome c oxidase


citrate synthase


endothelial nitric oxide synthase


inducible nitric oxide synthase


muscle-type CK


myocyte-specific enhancer binding factor 2


mitochondrial transcription factor A


myosin heavy chain


myogenic differentiation factor D


normal muscle


nitric oxide synthase


nuclear respiratory factor


peroxisome proliferator-activated receptor gamma co-activator


PGC-related co-activator


sarcomeric mitochondrial CK


sarcoma tissue


tumor necrosis factor α


  1. 1.

    Wallimann, T., Wyss, M., Brdiczka, D., Nicolay, K. and Eppenberger, H.M. Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit’ for cellular energy homeostasis. Biochem. J. 281 (1992) 21–40.

  2. 2.

    Qin, W., Khuchua, Z., Cheng, J., Boero, J., Payne, R.M. and Strauss, A.W. Molecular characterization of the creatine kinases and some historical perspectives. Mol. Cell Biochem. 184 (1998) 153–167.

  3. 3.

    Edmondson, D.G. and Olson, E.N. Helix-loop-helix proteins as regulators of muscle-specific transcription. J. Biol. Chem. 268 (1993) 755–758.

  4. 4.

    Martin, J.F., Schwarz, J.J. and Olson, E.N. Myocyte enhancer factor (MEF) 2C: A tissue-restricted member of the MEF-2 family of transcription factors. Proc. Natl. Acad. Sci. 90 (1993) 5282–5286.

  5. 5.

    Buskin, J.N. and Hauschka, S.D. Identification of a myocyte nuclear factor that binds to the muscle-specific enhancer of the mouse muscle creatine kinase gene. Mol. Cell Biol. 9 (1989) 2627–2640.

  6. 6.

    MoIkentin, J.D., Black, B.L., Martin, J.F. and Olson, E.N. Cooperative activation of muscle gene expression by MEF2 and myogenic bHLH proteins. Cell 83 (1995) 1125–1136.

  7. 7.

    Patra, S., Bera, S., Roy, S.S., Ghoshal, S., Ray, S., Basu, A., Schlattner, U., Wallimann, T. and Ray, M. Progressive decrease of phospho-creatine, creatine and creatine kinase in skeletal muscle upon transformation to sarcoma. FEBS J. 275 (2008) 3236–3247.

  8. 8.

    Langen, R.C., Van Der Velden, J.L., Schols, A.M., Kelders, M.C., Wouters, E.F. and Janssen-Heininger, Y.M. Tumor necrosis factor-alpha inhibits myogenic differentiation through MyoD protein destabilization. FASEB J. 18 (2004) 227–237.

  9. 9.

    Guttridge, D.C., Mayo, M.W., Madrid, L.V., Wang, C.Y. and Baldwin, A.S. Jr. NF-kappaB-induced loss of MyoD messenger RNA: possible role in muscle decay and cachexia. Science 289 (2000) 2363–2366.

  10. 10.

    Williams, G., Brown, T., Becker, L., Prager, M. and Giroir, B.P. Cytokine-induced expression of nitric oxide synthase in C2C12 skeletal muscle myocytes. Am. J. Physiol. 267 (1994) R1020–R1025.

  11. 11.

    Di Marco, S., Mazroui, R., Dallaire, P., Chittur, S., Tenenbaum, S.A., Radzioch, D., Marette, A. and Gallouzi, I.E. NF-kappa B-mediated MyoD decay during muscle wasting requires nitric oxide synthase mRNA stabilization, HuR protein, and nitric oxide release. Mol. Cell Biol. 25 (2005) 6533–6545.

  12. 12.

    Broussard, S.R., McCusker, R.H., Novakofski, J.E., Strle, K., Shen, W.H., Johnson, R.W., Freund, G.G., Dantzer, R. and Kelley, K.W. Cytokine-hormone interactions: tumor necrosis factor-α impairs biologic activity and downstream activation signals of the insulin-like growth factor I receptor in myoblasts. Endocrinology 144 (2003) 2988–2996.

  13. 13.

    Vaarmann, A., Fortin, D., Veksler, V., Momken, I., Ventura-Clapier, R. and Garnier, A. Mitochondrial biogenesis in fast skeletal muscle of CK deficient mice. Biochim. Biophys. Acta 1777 (2008) 39–47.

  14. 14.

    Scarpulla, R.C. Transcriptional paradigms in mammalian mitochondrial biogenesis and function. Physiol. Rev. 88 (2008) 611–638.

  15. 15.

    Scarpulla, R.C. Nuclear control of respiratory chain expression in mammalian cells. J. Bioenerg. Biomembr. 29 (1997) 109–119.

  16. 16.

    Moyes, C.D., Mathieu-costello, O.A., Tsuchiya, N., Filburn, C. and Hansford, R.G. Mitochondrial biogenesis during cellular differentiation. Am. J. Physiol. 272 (1997) Cl345–Cl351.

  17. 17.

    Leek, B.T., Mudaliar, S.R.D., Henry, R., Mathieu-costello, O.A. and Richardson, R.S. Effect of acute exercise on citrate synthase activity in untrained and trained human skeletal muscle. Am. J. Physiol. Regul. Integr. Comp. Physiol. 280 (2001) R441–R447.

  18. 18.

    Cox, B. and Emili, A. Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomics. Nat. Protoc. 1 (2006) 1872–1878.

  19. 19.

    Layne, E. Sepctrophotometric and turbidimetric methods for measuring proteins. Methods Enzymol. 3 (1957) 447–454.

  20. 20.

    Ross, J.A. and Nesnow, S. Polycyclic aromatic hydrocarbons: correlations between DNA adducts and ras oncogene mutations. Mutat. Res. 424 (1999) 155–166.

  21. 21.

    Kuru, S., Inukai, A., Kato, T., Liang, Y., Kimura, S. and Sobue, G. Expression of tumor necrosis factor-alpha in regenerating muscle fibers in inflammatory and noninflammatory myopathies. Acta Neuropathol. 105 (2003) 217–224.

  22. 22.

    Florini, J.R., Ewton, D.Z. and Coolican, S.A. Growth hormone and the insulin-like growth factor system in myogenesis. Endocr. Rev. 17 (1996) 481–517.

  23. 23.

    Yu, H. and Rohan, T. Role of the insulin-like growth factor family in cancer development and progression. J. Natl. Cancer Inst. 92 (2000) 1472–1489.

  24. 24.

    Frost, R.A., Nystrom, G.J. and Lang, C.H. Tumor necrosis factor-α decreases insulin-like growth factor-I messenger ribonucleic acid expression in C2C12 myoblasts via a Jun N-terminal kinase pathway. Endocrinology 144 (2003) 1770–1779.

  25. 25.

    Nisoli, E., Clementi, E., Paolucci, C., Cozzi, V., Tonello, C., Sciorati, C., Bracale, R., Valerio, A., Francolini, M., Moncada, S. and Carruba, M.O. Mitochondrial biogenesis in mammals: the role of endogenous nitric oxide. Science 299 (2003) 896–899.

  26. 26.

    Savagner, F., Mirebeau, D., Jacques, C., Guyetant, S., Morgan, C., Franc, B., Reynier, P. and Malthièry, Y. PGC-1-related coactivator and targets are upregulated in thyroid oncocytoma. Biochem. Biophys. Res. Commun. 310 (2003) 779–784.

  27. 27.

    Dong, X., Ghoshal, K., Majumder, S., Yadav, S.P. and Jacob, S.T. Mitochondrial transcription factor A and its downstream targets are upregulated in a rat hepatoma. J. Biol. Chem. 277 (2002) 43309–43318.

  28. 28.

    Kraft, C.S., LeMoine, C.M.R., Lyons, C.N., Michaud, D., Mueller, C.R. and Moyes, C.D. Control of mitochondrial biogenesis during myogenesis. Am. J. Physiol. Cell Physiol. 290 (2006) 1119–1127.

  29. 29.

    Handschin, C and Spiegelman, B.M. Peroxisome proliferator-activated receptor-γ coactivator 1 coactivators, energy homeostasis and metabolism. Endocr. Rev. 27 (2006) 728–735.

  30. 30.

    Chang, J.H., Lin, H.K., Chung, H.S., Chang, Y.J., Chi, H.C. and Chen, S.L. Myogenic bHLH proteins regulate the expression of PPARγ coactivator-1α. Endocrinology 147 (2006) 3093–3106.

  31. 31.

    Andersson, U. and Scarpulla, R.C. Pgc-1-related coactivator, a novel serum-inducible coactivator of nuclear respiratory factor 1-dependent transcription in mammalian cells, Mol. Cell Biol. 21 (2001) 3738–3749.

  32. 32.

    Ventura-Clapier, R., Kaasik, A. and Veksler, V. Structural and functional adaptations of striated muscles to CK deficiency. Mol. Cell Biochem. 256/257 (2004) 29–41.

  33. 33.

    de Groof, A.J.C., Oerlemans, F., Jost, C.R. and Wieringa, B. Changes in glycolytic network and mitochondrial design in creatine kinase-deficient muscles. Muscle Nerve 24 (2001) 1188–1196.

Download references

Author information



Corresponding author

Correspondence to Soumen Bera.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bera, S., Ray, M. The transcriptional cascade associated with creatine kinase down-regulation and mitochondrial biogenesis in mice sarcoma. Cell Mol Biol Lett 14, 481–496 (2009).

Download citation

Key words

  • Sarcoma
  • Creatine kinase
  • Nitric oxide synthase
  • Muscle degeneration
  • Mitochondrial biogenesis