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Cellular and Molecular Biology Letters

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1689-1392
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Volume 20, Issue 4

Ras Transformation Overrides a Proliferation Defect Induced by Tpm3.1 Knockout

Jason D. Coombes
  • Oncology Research Unit, School of Medical Sciences, UNSW Australia, Sydney, NSW 2052, Australia
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/ Galina Schevzov
  • Oncology Research Unit, School of Medical Sciences, UNSW Australia, Sydney, NSW 2052, Australia
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/ Chin-Yi Kan
  • Cancer Cell Development Group, Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, NSW 2052, Australia
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/ Carlotta Petti
  • Cancer Cell Development Group, Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, NSW 2052, Australia
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/ Michelle F. Maritz
  • Cancer Cell Development Group, Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, NSW 2052, Australia
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/ Shane Whittaker
  • Oncology Research Unit, School of Medical Sciences, UNSW Australia, Sydney, NSW 2052, Australia
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/ Karen L. Mackenzie
  • Cancer Cell Development Group, Children's Cancer Institute Australia, Lowy Cancer Research Centre, UNSW Australia, Sydney, NSW 2052, Australia
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/ Peter W. Gunning
  • Corresponding author
  • Oncology Research Unit, School of Medical Sciences, UNSW Australia, Sydney, NSW 2052, Australia
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Published Online: 2015-10-15 | DOI: https://doi.org/10.1515/cmble-2015-0037

Abstract

Extensive re-organisation of the actin cytoskeleton and changes in the expression of its binding proteins is a characteristic feature of cancer cells. Previously we have shown that the tropomyosin isoform Tpm3.1, an integral component of the actin cytoskeleton in tumor cells, is required for tumor cell survival. Our objective was to determine whether cancer cells devoid of Tpm3.1 would evade the tumorgenic effects induced by H-Ras transformation. The tropomyosin isoform (Tpm) expression profile of a range of cancer cell lines (21) demonstrates that Tpm3.1 is one of the most broadly expressed Tpm isoform. Consequently, the contribution of Tpm3.1 to the transformation process was functionally evaluated. Primary embryonic fibroblasts isolated from wild type (WT) and Tpm3.1 knockout (KO) mice were transduced with retroviral vectors expressing SV40 large T antigen and an oncogenic allele of the H-Ras gene, H-RasV12, to generate immortalized and transformed WT and KO MEFs respectively. We show that Tpm3.1 is required for growth factor-independent proliferation in the SV40 large T antigen immortalized MEFs, but this requirement is overcome by H-Ras transformation. Consistent with those findings, we found that Tpm3.1 was not required for anchorage independent growth or growth of H-Ras-driven tumors in a mouse model. Finally, we show that pERK and Importin 7 protein interactions are significantly decreased in the SV40 large T antigen immortalized KO MEFs but not in the H-Ras transformed KO cells, relative to control MEFs. The data demonstrate that H-Ras transformation overrides a requirement for Tpm3.1 in growth factor-independent proliferation of immortalized MEFs. We propose that in the SV40 large T antigen immortalized MEFs, Tpm3.1 is partly responsible for the efficient interaction between pERK and Imp7 resulting in cell proliferation, but this is overidden by Ras transformation.

This article offers supplementary material which is provided at the end of the article.

Keywords : Actin; Cytoskeleton; Tropomyosin; Cancer; Tumor; ERK; Ras; Transformation

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About the article

Received: 2015-05-28

Accepted: 2015-07-30

Published Online: 2015-10-15

Published in Print: 2015-12-01


Citation Information: Cellular and Molecular Biology Letters, Volume 20, Issue 4, Pages 626–646, ISSN (Online) 1689-1392, DOI: https://doi.org/10.1515/cmble-2015-0037.

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