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

Surinder K. Batra, Horst F. Kern, Andrew J. Worlock, Richard S. Metzgar, Michael A. Hollingsworth

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

Full-length cDNA for the human mucin Muc 1 gene under the control of the β 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.

Original languageEnglish (US)
Pages (from-to)841-849
Number of pages9
JournalJournal of cell science
Volume100
Issue number4
StatePublished - Dec 1991

Keywords

  • Differentiation
  • Mucin
  • Transfection

ASJC Scopus subject areas

  • Cell Biology

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