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

Author:

Batra S.K.1,Kern H.F.1,Worlock A.J.1,Metzgar R.S.1,Hollingsworth M.A.1

Affiliation:

1. Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina.

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.

Publisher

The Company of Biologists

Subject

Cell Biology

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