Stable transfection of CHO cells with the c-myc gene results in increased proliferation rates, reduces serum dependency, and induces anchorage independence (original) (raw)

Abstract

Induction of the transcription factor Myc promotes cell proliferation and transformation by activating growth-promoting genes and/or by transcriptionally repressing the expression of growth arrest genes. However, a number of studies have shown that c-Myc is a potent inducer of apoptosis in the absence of serum or growth factors. To further examine the role of Myc in cell growth and proliferation, and the advantages of this positive regulator in cell culture we transfected the CHO-K1 cell line with a human c-myc gene driven by MMLV 5′-LTR promoter. Over-expression of ectopic c-Myc resulted in a significant increase in growth rate and maximum cell number, in both suspension and attached batch culture accompanied by a similar decrease in specific glucose consumption rate. Interestingly, there was no manifestation of the widely reported apoptotic death by c-myc in the absence of serum. Additionally, over-expression of c-Myc appeared to induce morphological transformation and partial anchorage-independence.

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References

• Al-Rubeai M., Singh R.P., Goldaman M.H. and Emery A.N. 1995. The death mechanism of animal cells in conditions of intensive agitation. Biotech Bioeng 45: 463–472.
  Google Scholar
• Al-Rubeai M. 1998. Apoptosis and cell culture technology. In: Scheper T. (ed.), Advances in Biochemical Engineering/Biotechnology Vol. 59. Springer-Verlag, pp. 226–249.
• Amati B., Littlewood T.D., Evan G.I. and Land H. 1993. The c-Myc protein induces cell cycle progression and apoptosis through dimerization with Max. EMBO J 13: 5083–5087.
  Google Scholar
• Bissonnette R.P., Echeverri F., Mahboubi A. and Green D.R. 1992. Apoptotic cell death induced by _c_-myc is inhibited by _bcl_-2. Nature 359: 552–554.
  Google Scholar
• Bouchard C., Staller P. and Eilers M. 1998. Control of cell proliferation by Myc. Trends Cell Biol 8: 202–206.
  Google Scholar
• Claasen G.F. and Hann S.R. 1999. Myc mediated transformation: the repression connection. Oncogene 18: 2925–2933.
  Google Scholar
• Cole M.D. 1986. The myc oncogenes: its role in transformation and differentiation. Ann Rev Genet 20: 361–384.
  Google Scholar
• Evan G.I., Wyllie A.H., Gilbert C.S., Littlewood T.D., Land H., Brooks M. et al. 1992. Induction of apoptosis in fibroblasts by c-Myc protein. Cell 69: 119–128.
  Google Scholar
• Evan G.I., Harrington E., McCarthy N., Gilbert C., Benedict M.A. and Nuñez G. 1996. Integrated control of cell proliferation and apoptosis by oncogenes. In: Thomas N.S.B. (ed.), Apoptosis and Cell Cycle Control in Cancer. BIOS Scientific Publishers, Oxford, pp. 109–129.
  Google Scholar
• Facchini L.M. and Penn L.Z. 1998. The molecular role of Myc in growth and transformation: recent discoveries lead to new insights. FASEB J 12: 633–651.
  Google Scholar
• Fanidi A., Harrington E.A. and Evan G.I. 1992. cooperative interaction between _c_-myc and _bcl_-2 proto-oncogenes. Nature 359: 554–556.
  Google Scholar
• Fussenegger M. and Bailey J.E. 1998. Molecular regulation of cell-cycle progression and apoptosis in mammalian cells: implications for biotechnology. Biotechnol Prog 14: 807–833.
  Google Scholar
• Hoffman B. and Liebermann D.A. 1998. The proto-oncogene _c_-myc and apoptosis. Oncogene 17: 3351–3357.
  Google Scholar
• Marcu K.B., Bossonne S.A. and Patel A.J. 1992. Myc function and regulation. Annu Rev Biochem 61: 809–860.
  Google Scholar
• Meichle A., Philipp A. and Eilers M. 1992. The functions of Myc proteins. Biochem Biophys Acta 1114: 129–146.
  Google Scholar
• Penn L.J.Z., Brooks M.W., Laufer E.M. and Land H. 1990. Negative autoregulation of _c_-myc transcription. EMBO J 9: 1113–1121.
  Google Scholar
• Prendergast G.C. 1999. Mechanisms of apoptosis by c-Myc. Oncogene 18: 2967–2987.
  Google Scholar
• Sanfeliou A. and Stephanopoulos G. 1999. Effect of glutamine limitation on the death of attached Chinese hamster ovary cells. Biotech Bioeng 64: 46–53.
  Google Scholar
• Simpson N.A., Milner E. and Al-Rubeai M. 1997. Prevention of hybridoma cell death by bcl-2 during sub-optimal culture conditions. Biotech Bioeng 54: 1–16.
  Google Scholar
• Small M.B., Hay N., Schwab M. and Bishop M. 1987. Neoplastic transformation by the human gene _N_-myc. Mol Cell Biol 7: 1638–1645.
  Google Scholar
• Spencer C.A. and Groudine M. 1991. Control of _c_-myc regulation in normal and neoplastic cells. Adv Cancer Res 56: 1–48.
  Google Scholar
• Wahgner A.J., Small M.B. and Hay N. 1993. Myc-mediated apoptosis is blocked by ectopic expression of Bcl-2. Mol Cell Biol 13: 2432–2440.
  Google Scholar

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Authors and Affiliations

1. Animal Cell Technology Group, Department of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, B15 2 TT, UK
   Vasiliki Ifandi & Mohamed Al-Rubeai

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1. Vasiliki Ifandi
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2. Mohamed Al-Rubeai
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Ifandi, V., Al-Rubeai, M. Stable transfection of CHO cells with the c-myc gene results in increased proliferation rates, reduces serum dependency, and induces anchorage independence.Cytotechnology 41, 1–10 (2003). https://doi.org/10.1023/A:1024203518501

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• Issue Date: January 2003
• DOI: https://doi.org/10.1023/A:1024203518501