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Related Experiment Videos

Transformation by v-Src: Ras-MAPK and PI3K-mTOR mediate parallel pathways.

E Penuel1, G S Martin

  • 1Department of Molecular and Cell Biology, University of California-Berkeley, Berkeley, California 94720-3204, USA.

Molecular Biology of the Cell
|June 8, 1999
PubMed
Summary
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v-Src transformation of chicken embryo fibroblasts relies on two parallel pathways: Ras-MAPK and PI3K-mTOR. Inhibiting either pathway alone is insufficient, but simultaneous blockade effectively prevents transformation, highlighting their combined role.

Area of Science:

  • Cell Biology
  • Molecular Oncology
  • Signal Transduction

Background:

  • v-Src mediated transformation of chicken embryo fibroblasts (CEF) is a complex process.
  • While active Ras (GTP-bound Ras) is not essential for transformation, suggesting alternative signaling routes.
  • The phosphatidylinositol-3-kinase (PI3K)-mammalian target of rapamycin (mTOR) pathway is a potential Ras-independent contributor.

Purpose of the Study:

  • To investigate the contribution of the PI3K-mTOR pathway to v-Src transformation.
  • To determine the combined effect of inhibiting both Ras-MAPK and PI3K-mTOR pathways on CEF transformation by v-Src.

Main Methods:

  • Transformation parameters assessed: morphological alteration, hexose uptake, density inhibition, anchorage-independent growth.
  • Ras-MAPK pathway inhibition using dominant-negative HRasN17 or MEK inhibitor PD98059.

Related Experiment Videos

  • PI3K-mTOR pathway inhibition using PI3K inhibitor LY294002 or mTOR inhibitor rapamycin.

    • Main Results:

    • Inhibition of Ras-MAPK or PI3K-mTOR pathways individually reduced transformation parameters but did not block it.
    • Simultaneous inhibition of both Ras-MAPK and PI3K-mTOR pathways effectively blocked v-Src-induced transformation.
    • These findings indicate two parallel, cooperating pathways are crucial for transformation.

    Conclusions:

    • v-Src transformation of CEF is mediated by two parallel signaling pathways: Ras-MAPK and PI3K-mTOR.
    • Both pathways are necessary and contribute significantly to the transformation process.
    • Targeting both pathways concurrently offers a potential strategy to inhibit v-Src-driven transformation.