Transfection of the human Muc 1 mucin gene into a poorly differentiated human pancreatic tumor cell line, Panc1: integration, expression and ultrastructural changes.
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.
Batra
S K
SK
Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina.
Kern
H F
HF
Worlock
A J
AJ
Metzgar
R S
RS
Hollingsworth
M A
MA
D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002454:Cell Differentiation; D003593:Cytoplasm / Q000648:ultrastructure; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm / Q000235:genetics*; D006056:Golgi Apparatus / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000235:genetics*; D008854:Microscopy, Electron; D018396:Mucin-1; D009077:Mucins / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D010179:Pancreas / Q000166:cytology / Q000648:ultrastructure*; D014176:Protein Biosynthesis; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D008562:Membrane Glycoproteins; D018396:Mucin-1; D009077:Mucins; D009363:Neoplasm Proteins