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Related Experiment Videos

Changes in gap junctional intercellular communication in mouse skin carcinogenesis

P R Holden1, B McGuire, A Stoler

  • 1The Beatson Institute for Cancer Research, CRC Beatson Laboratories, Bearsden, Glasgow, UK.

Carcinogenesis
|January 1, 1997
PubMed
Summary

Gap junction intercellular communication (GJIC) remains unchanged in early skin cancer stages but significantly decreases in spindle carcinoma cells. E-cadherin loss, not connexin 43, is linked to this communication defect.

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Area of Science:

  • Cell Biology
  • Cancer Research
  • Molecular Biology

Background:

  • Gap junction intercellular communication (GJIC) is vital for tissue homeostasis.
  • Dysregulation of GJIC is implicated in various cancers.
  • Chemically induced mouse skin carcinogenesis provides a model to study GJIC alterations during tumor progression.

Purpose of the Study:

  • To investigate changes in GJIC during different stages of mouse skin carcinogenesis.
  • To identify molecular factors associated with decreased GJIC in advanced skin cancer.
  • To explore the role of E-cadherin and connexin 43 in GJIC regulation during tumor progression.

Main Methods:

  • Measurement of GJIC using dye spread assays in various cell lines.
  • Analysis of Ha-ras gene mutations.

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  • Somatic cell hybridization for genetic complementation studies.
  • Western blotting and immunofluorescence to assess protein expression (connexin 43, E-cadherin).
  • Gene transfection to restore E-cadherin expression.

    • Main Results:

    • GJIC levels were similar in normal keratinocytes, papillomas, and squamous carcinomas.
    • A significant 80-90% decrease in GJIC was observed in spindle carcinoma cells compared to squamous carcinoma cells.
    • The defect causing reduced GJIC in spindle cells was recessive and linked to E-cadherin deficiency, not connexin 43 abnormalities.
    • Restoration of E-cadherin partially rescued GJIC in spindle cells.
    • A separate, non-tumorigenic keratinocyte line showed a distinct GJIC defect with abnormal connexin 43 distribution.

    Conclusions:

    • Loss of GJIC is a late event in skin carcinogenesis, specifically occurring during the transition to spindle cell carcinoma.
    • E-cadherin plays a crucial role in maintaining GJIC in skin epithelial cells.
    • The genetic defect in spindle carcinoma cells involves E-cadherin, while other keratinocyte lines may have different GJIC regulatory mechanisms.