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Recombinant interferon-gamma differently affects DQA and DRA gene expression in two human melanoma clones:

C Castelli1, M Sensi, G Parmiani

  • 1Division of Experimental Oncology D, Istituto Nazionale per lo Studio e la Cura dei Tumori, Milan, Italy.

Experimental and Clinical Immunogenetics
|January 1, 1991
PubMed

Insights

Recombinant interferon gamma (rINF-gamma) differentially regulates human leukocyte antigen (HLA) DQ gene expression in melanoma. This regulation occurs at both transcriptional and posttranscriptional levels, involving a short-lived protein stabilizing DQ mRNA.

Area of Science:

  • Immunogenetics
  • Molecular Biology
  • Cancer Research

Background:

  • Human melanoma cells exhibit differential responses to recombinant interferon gamma (rINF-gamma).
  • Previous studies showed distinct HLA class II gene expression patterns in two melanoma clones (9229/18 and 9229/5) upon rINF-gamma treatment.
  • Clone 9229/18 showed coordinate DR and DQ gene expression, while clone 9229/5 showed DR but not DQ gene upregulation.

Purpose of the Study:

  • To investigate the molecular mechanisms behind differential HLA DQ gene regulation by rINF-gamma in human melanoma.
  • To analyze DQA gene regulation at both transcriptional and posttranscriptional levels.
  • To identify factors influencing DQA mRNA stability.

Main Methods:

  • Nuclear run-on assays to assess gene transcription rates.
  • Analysis of steady-state mRNA levels and cell surface antigen expression.
  • mRNA turnover studies using actinomycin D and cycloheximide to evaluate mRNA stability.
  • Treatment of melanoma cell clones with rINF-gamma.

Main Results:

  • rINF-gamma induced coordinate transcriptional activation of DRA and DQA genes in clone 9229/18.
  • In clone 9229/5, rINF-gamma strongly increased DRA transcription but not DQA transcription.
  • A short-lived protein was found to be essential for stabilizing human DQA mRNA in melanoma cells.
  • Basal transcription rates for class II genes did not yield mature cytoplasmic mRNA in either clone.

Conclusions:

  • DQA gene expression in melanoma is tightly regulated at both transcriptional and posttranscriptional levels.
  • rINF-gamma can differentially regulate DQA and DRA genes transcriptionally in specific melanoma clones.
  • Posttranscriptional regulation of DQA mRNA stability is mediated by a short-lived protein, highlighting complex gene expression control in melanoma.

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