Cooperative transformation of 32D cells by the combined expression of IRS-1 and V-Ha-Ras

Barbara Cristofanelli, Barbara Valentinis, Silvia Soddu, Maria Giulia Rizzo, Alessandra Marchetti, Gianluca Bossi, Anna Rita Morena, Michael Dews, Renato Baserga, Ada Sacchi

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

32D cells expressing v-Ha-Ras fail to show a transformed phenotype. Since Ras requires an active IGF-1R for transformation of fibroblasts, we asked whether expression of IRS-1 or Shc (two of the major substrates of the IGF-1R) could co-operate with oncogenic Ras in transforming 32D cells. We find that IRS-1, but not Shc, in combination with v-Ha-Ras generates a fully transformed phenotype in 32D cells. 32D cells expressing both IRS-1 and v-Ha-Ras (32D/IRS1/Ras) survive and proliferate in the absence of IL-3, do not undergo granulocytic differentiation in the presence of G-CSF and form tumors in nu/nu and syngeneic mice. In contrast, 32D cells expressing singly IRS-1 or v-Ha-Ras exhibit only a block in differentiation capacity. Over-expression of Shc proteins, by itself, promotes differentiation of 32D cells. Concomitant expression of IRS-1 and v-Ha-Ras synergistically phosphorylates ERK-1 and ERK-2 whereas a MEK inhibitor rapidly induces death of 32D/IRS1/Ras transformed cells. Furthermore, transformed 32D/IRS1/Ras cells display high levels of PI3-K activation and undergo rapid apoptosis when exposed to P13-K inhibitors. The data indicate that: (1) a fully transformed phenotype in 32D cells is generated when a block in differentiation (v-Ha-Ras) is coupled with another differentiation block (IRS-1); (2) PI3-K and MAPK activity are required for the survival of transformed cells; (3) the signals generated by IRS-1 and oncogenic Ras converge on ERK and P13-K resulting in high levels of activation.

Original languageEnglish
Pages (from-to)3245-3255
Number of pages11
JournalOncogene
Volume19
Issue number29
Publication statusPublished - Jul 6 2000

Fingerprint

Phenotype
Interleukin-3
Mitogen-Activated Protein Kinase Kinases
Granulocyte Colony-Stimulating Factor
Cell Differentiation
Cell Survival
Fibroblasts
Apoptosis
Neoplasms
Proteins

Keywords

  • MAPK
  • Myelodysplastic syndrome
  • PI3-K
  • Shc
  • Transformation

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Cristofanelli, B., Valentinis, B., Soddu, S., Rizzo, M. G., Marchetti, A., Bossi, G., ... Sacchi, A. (2000). Cooperative transformation of 32D cells by the combined expression of IRS-1 and V-Ha-Ras. Oncogene, 19(29), 3245-3255.

Cooperative transformation of 32D cells by the combined expression of IRS-1 and V-Ha-Ras. / Cristofanelli, Barbara; Valentinis, Barbara; Soddu, Silvia; Rizzo, Maria Giulia; Marchetti, Alessandra; Bossi, Gianluca; Morena, Anna Rita; Dews, Michael; Baserga, Renato; Sacchi, Ada.

In: Oncogene, Vol. 19, No. 29, 06.07.2000, p. 3245-3255.

Research output: Contribution to journalArticle

Cristofanelli, B, Valentinis, B, Soddu, S, Rizzo, MG, Marchetti, A, Bossi, G, Morena, AR, Dews, M, Baserga, R & Sacchi, A 2000, 'Cooperative transformation of 32D cells by the combined expression of IRS-1 and V-Ha-Ras', Oncogene, vol. 19, no. 29, pp. 3245-3255.
Cristofanelli B, Valentinis B, Soddu S, Rizzo MG, Marchetti A, Bossi G et al. Cooperative transformation of 32D cells by the combined expression of IRS-1 and V-Ha-Ras. Oncogene. 2000 Jul 6;19(29):3245-3255.
Cristofanelli, Barbara ; Valentinis, Barbara ; Soddu, Silvia ; Rizzo, Maria Giulia ; Marchetti, Alessandra ; Bossi, Gianluca ; Morena, Anna Rita ; Dews, Michael ; Baserga, Renato ; Sacchi, Ada. / Cooperative transformation of 32D cells by the combined expression of IRS-1 and V-Ha-Ras. In: Oncogene. 2000 ; Vol. 19, No. 29. pp. 3245-3255.
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