Transfection of the human Muc 1 mucin gene into a poorly differentiated human pancreatic tumor cell line, Panc1: integration, expression and ultrastructural changes

J Cell Sci. 1991 Dec;100 ( Pt 4):841-9. doi: 10.1242/jcs.100.4.841.

Abstract

Full-length cDNA for the human mucin Muc 1 gene under the control of the beta actin promoter was transfected into a morphologically poorly differentiated pancreatic tumor cell line, Panc 1, by the DEAE-dextran method. Integration of the foreign Muc 1 cDNA occurred at multiple sites in the genome of Panc 1. Northern blot analysis showed Muc 1 expression in cells transfected with the Muc 1 cDNA, but not in control cells transfected with vector alone or an antisense Muc 1 cDNA construct. Transfection of Panc 1 with Muc 1 cDNA did not cause any detectable alteration or rearrangements in the Muc 1 gene or cDNA. Western blot analysis of cell lysates from the transfected lines using a monoclonal antibody reactive with the Muc 1 protein (HMFG-2) demonstrated that Muc 1 protein expression correlated with the Northern blot data. Immunoperoxidase staining using HMFG-2 showed that Muc 1 protein was expressed in less than 5% of control Panc 1 cells, whereas greater than 95% of cells transfected with Muc 1 cDNA expressed the protein. Ultrastructural examination of Muc 1-transfected cells demonstrated the formation of dense core granules and increased amounts of rough endoplasmic reticulum.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Blotting, Northern
  • Blotting, Southern
  • Blotting, Western
  • Cell Differentiation
  • Cytoplasm / ultrastructure
  • Gene Expression Regulation, Neoplastic*
  • Genes, Neoplasm / genetics*
  • Golgi Apparatus / ultrastructure
  • Humans
  • Immunohistochemistry
  • Membrane Glycoproteins / genetics*
  • Microscopy, Electron
  • Mucin-1
  • Mucins / genetics*
  • Neoplasm Proteins / genetics*
  • Pancreas / cytology
  • Pancreas / ultrastructure*
  • Protein Biosynthesis
  • Transcription, Genetic
  • Transfection*
  • Tumor Cells, Cultured

Substances

  • Membrane Glycoproteins
  • Mucin-1
  • Mucins
  • Neoplasm Proteins