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Altered folate-binding protein mRNA stability in KB cells grown in folate-deficient medium

C T Hsueh1, B J Dolnick

  • 1Department of Experimental Therapeutics, Grace Cancer Drug Center, Roswell Park Cancer Institute, Buffalo, NY 14263.

Biochemical Pharmacology
|June 22, 1993
PubMed

Insights

Folate-binding protein (FBP) mRNA levels increase in folate-deficient cells. This rise is primarily due to enhanced mRNA stability, not increased gene expression or methylation changes.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Folate-binding protein (FBP) facilitates cellular uptake of folate and analogs like methotrexate.
  • FBP expression is significantly upregulated in human KB cells under folate-deficient conditions.

Purpose of the Study:

  • To investigate the mechanisms behind enhanced FBP gene expression in folate-deficient KB cells.
  • To determine if gene organization, copy number, methylation, transcription, or mRNA stability contribute to increased FBP levels.

Main Methods:

  • Southern blot analysis to assess FBP gene organization and copy number.
  • DNA methylation analysis of FBP gene in folate-replete and deficient cells.
  • Assessment of FBP gene transcriptional rates.
  • RNA half-life studies to evaluate FBP mRNA stability.

Main Results:

  • No significant alterations in FBP gene organization or copy number were observed.
  • Folate deficiency led to FBP gene hypomethylation, but methylation inhibitors did not increase FBP mRNA.
  • Transcriptional rates of the FBP gene remained constant between conditions.
  • FBP mRNA half-life was approximately 2.5-fold longer in folate-deficient cells.

Conclusions:

  • The increased FBP mRNA levels in folate-deficient KB cells are primarily attributed to enhanced mRNA stability.
  • Mechanisms involving gene organization, copy number, and direct transcriptional regulation do not explain the observed upregulation.

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