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

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

Research output: Contribution to journalArticle

38 Citations (Scopus)

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

Fingerprint

Mucin-1
Tumor Cell Line
Transfection
Complementary DNA
Genes
Northern Blotting
Proteins
DEAE-Dextran
Rough Endoplasmic Reticulum
Actins
Western Blotting
Monoclonal Antibodies
Genome
Staining and Labeling

Keywords

  • Differentiation
  • Mucin
  • Transfection

ASJC Scopus subject areas

  • Cell Biology

Cite this

@article{bf11e937d0f84843ac74197e5036b064,
title = "Transfection of the human Muc 1 mucin gene into a poorly differentiated human pancreatic tumor cell line, Panc1: Integration, expression and ultrastructural changes",
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.",
keywords = "Differentiation, Mucin, Transfection",
author = "Batra, {Surinder Kumar} and Kern, {Horst F.} and Worlock, {Andrew J.} and Metzgar, {Richard S.} and Hollingsworth, {Michael A}",
year = "1991",
month = "12",
language = "English (US)",
volume = "100",
pages = "841--849",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "4",

}

TY - JOUR

T1 - Transfection of the human Muc 1 mucin gene into a poorly differentiated human pancreatic tumor cell line, Panc1

T2 - Integration, expression and ultrastructural changes

AU - Batra, Surinder Kumar

AU - Kern, Horst F.

AU - Worlock, Andrew J.

AU - Metzgar, Richard S.

AU - Hollingsworth, Michael A

PY - 1991/12

Y1 - 1991/12

N2 - 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.

AB - 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.

KW - Differentiation

KW - Mucin

KW - Transfection

UR - http://www.scopus.com/inward/record.url?scp=0026349593&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026349593&partnerID=8YFLogxK

M3 - Article

C2 - 1814933

AN - SCOPUS:0026349593

VL - 100

SP - 841

EP - 849

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 4

ER -