Expression of the cystic fibrosis transmembrane conductance regulator gene in cells of non-epithelial origin. Academic Article uri icon

Overview

MeSH

  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cell Line
  • Cells, Cultured
  • Chromosome Mapping
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Deoxyribonuclease I
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Tumor Cells, Cultured

MeSH Major

  • Cystic Fibrosis
  • Gene Expression
  • Membrane Proteins

abstract

  • Consistent with the fact that the clinical disorder cystic fibrosis (CF) is manifested on epithelial surfaces, active transcription of the CF transmembrane conductance regulator (CFTR) gene and CFTR mRNA transcripts are detectable in a variety of epithelial cells, suggesting CFTR gene expression might be epithelial cell-specific. However, analysis of the CFTR gene promoter suggests it is a housekeeping gene, implying more widespread expression than only in epithelial cells. To evaluate the latter hypothesis, various human cells of non-epithelial origin, including lung fibroblasts, U-937 histiocytic lymphoma cells, K-562 erythroleukemia cells, HL-60 promyelocytic leukemia cells as well as freshly isolated blood lymphocytes, neutrophils, monocytes, and alveolar macrophages were examined for CFTR gene expression. Although Northern analysis failed to show CFTR mRNA transcripts in these cells, amplification of mRNA (after conversion to cDNA) by polymerase chain reaction combined with Southern analysis demonstrated the presence of CFTR mRNA transcripts at low levels in all cells evaluated except HL-60 cells. Comparative quantitative analysis showed fibroblasts contained 200-400 fold less CFTR mRNA transcripts than the T84 and HT-29 colon carcinoma epithelial cell lines, but had similar levels of CFTR transcripts to those of other epithelial cell lines. Nuclear transcription run-on analyses demonstrated very low level CFTR gene transcription in fibroblasts and U-937 cells, similar to that of other epithelial cells, but lower than the T84 and HT-29 colon carcinoma cell lines. Interestingly, while chromatin DNA of fibroblasts had no DNase I hypersensitivity sites in the 5' flanking region of the CFTR gene, HT-29 chromatin DNA exhibited four DNase I accessible sites in the same region, suggesting that these sites may be related to more active transcription of the CFTR gene in the intestinal epithelial cells than in fibroblasts.

publication date

  • October 11, 1991

has subject area

  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cell Line
  • Cells, Cultured
  • Chromosome Mapping
  • Cystic Fibrosis
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Deoxyribonuclease I
  • Gene Expression
  • Humans
  • Membrane Proteins
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Tumor Cells, Cultured

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC328907

PubMed ID

  • 1717947

Additional Document Info

start page

  • 5417

end page

  • 5423

volume

  • 19

number

  • 19