Anchorage-independent growth of pocket protein-deficient murine fibroblasts requires bypass of G2 arrest and can be accomplished by expression of TBX2

Mol Cell Biol. 2008 Dec;28(24):7263-73. doi: 10.1128/MCB.00313-08. Epub 2008 Oct 20.

Abstract

Mouse embryonic fibroblasts (MEFs) deficient for pocket proteins (i.e., pRB/p107-, pRB/p130-, or pRB/p107/p130-deficient MEFs) have lost proper G(1) control and are refractory to Ras(V12)-induced senescence. However, pocket protein-deficient MEFs expressing Ras(V12) were unable to exhibit anchorage-independent growth or to form tumors in nude mice. We show that depending on the level of pocket proteins, loss of adhesion induces G(1) and G(2) arrest, which could be alleviated by overexpression of the TBX2 oncogene. TBX2-induced transformation occurred only in the absence of pocket proteins and could be attributed to downregulation of the p53/p21(CIP1) pathway. Our results show that a balance between the pocket protein and p53 pathways determines the level of transformation of MEFs by regulating cyclin-dependent kinase activities. Since transformation of human fibroblasts also requires ablation of both pathways, our results imply that the mechanisms underlying transformation of human and mouse cells are not as different as previously claimed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Transformation, Neoplastic
  • Cyclin-Dependent Kinase Inhibitor p21 / genetics
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Fibroblasts / cytology
  • Fibroblasts / physiology*
  • G2 Phase / physiology*
  • Humans
  • Mice
  • Mice, Knockout
  • Mice, Nude
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Retinoblastoma Protein / genetics
  • Retinoblastoma Protein / metabolism*
  • Retinoblastoma-Like Protein p107 / genetics
  • Retinoblastoma-Like Protein p107 / metabolism*
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / metabolism*
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism
  • ras Proteins / genetics
  • ras Proteins / metabolism

Substances

  • Cdkn1a protein, mouse
  • Cyclin-Dependent Kinase Inhibitor p21
  • Protein Isoforms
  • Retinoblastoma Protein
  • Retinoblastoma-Like Protein p107
  • T-Box Domain Protein 2
  • T-Box Domain Proteins
  • Tumor Suppressor Protein p53
  • ras Proteins