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Related Experiment Videos

Decrease in epidermal growth factor receptor levels and production of material enhancing epidermal growth factor

P Guinivan, R L Ladda

    Proceedings of the National Academy of Sciences of the United States of America
    |July 1, 1979
    PubMed
    Summary

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    Temperature-sensitive (Ts) cells infected with Kirsten sarcoma virus showed reversible transformation, correlating with epidermal growth factor (EGF) receptor levels and production of growth-stimulating substances.

    Area of Science:

    • Cell Biology
    • Virology
    • Cancer Research

    Background:

    • Normal rat kidney (NRK) cells exhibit normal morphology and respond to epidermal growth factor (EGF).
    • Transformation of NRK cells by Kirsten sarcoma virus (KNRK) alters morphology and EGF responsiveness.
    • Temperature-sensitive (Ts) mutants of Kirsten sarcoma virus allow for reversible studies of viral transformation.

    Purpose of the Study:

    • To investigate the relationship between the transformed phenotype induced by a temperature-sensitive Kirsten sarcoma virus mutant and epidermal growth factor (EGF) receptor levels.
    • To characterize the substances produced by transformed cells that influence cell growth and EGF receptor expression.

    Main Methods:

    • Culturing normal rat kidney (NRK) cells, KNRK cells, and Ts cells at permissive (32°C) and nonpermissive (39°C) temperatures.

    Related Experiment Videos

  • Assessing cell morphology and growth patterns (monolayer vs. multilayer foci).
  • Measuring epidermal growth factor (EGF) binding using 125I-labeled EGF to determine EGF receptor levels.
  • Performing crossfeeding experiments with conditioned medium to evaluate the production of growth-stimulating substances.

    • Main Results:

    • Ts cells displayed normal morphology at 39°C and transformed morphology at 32°C, mirroring KNRK cells.
    • NRK cells showed high EGF receptor levels and responded to EGF, while KNRK cells lacked detectable EGF receptors and were unresponsive to EGF.
    • Ts cells had reduced EGF binding, which varied with temperature, and produced heat-stable substances that stimulated NRK cell DNA synthesis.
    • Conditioned medium from transformed cells enhanced EGF binding to NRK cells without competing for the EGF receptor.

    Conclusions:

    • A direct correlation exists between the transformed phenotype and epidermal growth factor (EGF) receptor levels, which is reversible in Ts cells.
    • Transformed cells produce novel factors that stimulate DNA synthesis and enhance EGF binding, independent of EGF competition.
    • These findings offer insights into the mechanisms of viral transformation and cell growth regulation.