Smokeless tobacco extracts modulate exogenous gene expression in early passage cultured human oral epithelial cells - An in vitro system to study chemical and viral enhancer/promoter interaction

Cem S. Demirci, Don R. Miller, Jacques Baillargeon, Mary Pat Moyer, Hal B. Jenson

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The increased risk for cancers of the oral cavity from smokeless tobacco use may reflect the interaction of tobacco with genetic factors, such as oncogenes, and other exogenous factors, such as viruses. An in vitro system was developed based on expression of the chloramphenicol acetyltransferase (CAT) reporter gene to study interactions of chemical treatments with viral enhancer/promoters in early passage cell cultures of oral cavity-derived epithelial cells. Expression of CAT in transfected cells was significantly greater with CAT under the control of the cytomegalovirus immediate early enhancer/promoter (pCEP4/CAT) compared to the Rous sarcoma virus long terminal repeat enhancer/promoter (pRSV-cat) and the simian virus 40 (SV40) early promoter (pSV2-cat). No CAT expression was detected using corresponding control plasmids without the CAT reporter gene. Using this system, smokeless tobacco extracts prepared from either dry snuff or moist snuff delayed maximum CAT expression from Day 4 to Day 5, with sustained, significantly increased CAT expression at 12 days compared to the declining CAT expression observed in untreated control cells. Smokeless tobacco extracts can modulate intracellular gene expression. This system provides an in vitro model to test specificity of toxic agents on enhancer/promoter activity and the interaction on exogenous gene expression.

Original languageEnglish (US)
Pages (from-to)527-532
Number of pages6
JournalJournal of Pharmacological and Toxicological Methods
Volume44
Issue number3
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Smokeless Tobacco
Chloramphenicol O-Acetyltransferase
Gene expression
Epithelial Cells
Gene Expression
Viruses
Reporter Genes
Mouth
Cats
Genes
In Vitro Techniques
Rous sarcoma virus
Simian virus 40
Tobacco
Terminal Repeat Sequences
Poisons
Mouth Neoplasms
Tobacco Use
Cytomegalovirus
Oncogenes

Keywords

  • Chloramphenicol acetyltransferase (CAT)
  • Enhancer/promoter
  • Gene expression
  • Smokeless tobacco

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

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title = "Smokeless tobacco extracts modulate exogenous gene expression in early passage cultured human oral epithelial cells - An in vitro system to study chemical and viral enhancer/promoter interaction",
abstract = "The increased risk for cancers of the oral cavity from smokeless tobacco use may reflect the interaction of tobacco with genetic factors, such as oncogenes, and other exogenous factors, such as viruses. An in vitro system was developed based on expression of the chloramphenicol acetyltransferase (CAT) reporter gene to study interactions of chemical treatments with viral enhancer/promoters in early passage cell cultures of oral cavity-derived epithelial cells. Expression of CAT in transfected cells was significantly greater with CAT under the control of the cytomegalovirus immediate early enhancer/promoter (pCEP4/CAT) compared to the Rous sarcoma virus long terminal repeat enhancer/promoter (pRSV-cat) and the simian virus 40 (SV40) early promoter (pSV2-cat). No CAT expression was detected using corresponding control plasmids without the CAT reporter gene. Using this system, smokeless tobacco extracts prepared from either dry snuff or moist snuff delayed maximum CAT expression from Day 4 to Day 5, with sustained, significantly increased CAT expression at 12 days compared to the declining CAT expression observed in untreated control cells. Smokeless tobacco extracts can modulate intracellular gene expression. This system provides an in vitro model to test specificity of toxic agents on enhancer/promoter activity and the interaction on exogenous gene expression.",
keywords = "Chloramphenicol acetyltransferase (CAT), Enhancer/promoter, Gene expression, Smokeless tobacco",
author = "Demirci, {Cem S.} and Miller, {Don R.} and Jacques Baillargeon and Moyer, {Mary Pat} and Jenson, {Hal B.}",
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T1 - Smokeless tobacco extracts modulate exogenous gene expression in early passage cultured human oral epithelial cells - An in vitro system to study chemical and viral enhancer/promoter interaction

AU - Demirci, Cem S.

AU - Miller, Don R.

AU - Baillargeon, Jacques

AU - Moyer, Mary Pat

AU - Jenson, Hal B.

PY - 2000

Y1 - 2000

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AB - The increased risk for cancers of the oral cavity from smokeless tobacco use may reflect the interaction of tobacco with genetic factors, such as oncogenes, and other exogenous factors, such as viruses. An in vitro system was developed based on expression of the chloramphenicol acetyltransferase (CAT) reporter gene to study interactions of chemical treatments with viral enhancer/promoters in early passage cell cultures of oral cavity-derived epithelial cells. Expression of CAT in transfected cells was significantly greater with CAT under the control of the cytomegalovirus immediate early enhancer/promoter (pCEP4/CAT) compared to the Rous sarcoma virus long terminal repeat enhancer/promoter (pRSV-cat) and the simian virus 40 (SV40) early promoter (pSV2-cat). No CAT expression was detected using corresponding control plasmids without the CAT reporter gene. Using this system, smokeless tobacco extracts prepared from either dry snuff or moist snuff delayed maximum CAT expression from Day 4 to Day 5, with sustained, significantly increased CAT expression at 12 days compared to the declining CAT expression observed in untreated control cells. Smokeless tobacco extracts can modulate intracellular gene expression. This system provides an in vitro model to test specificity of toxic agents on enhancer/promoter activity and the interaction on exogenous gene expression.

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