Relative promoter strengths in four human prostate cancer cell lines evaluated by particle bombardment-mediated gene transfer

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Abstract

BACKGROUND. The particle bombardment (gene gun) method for gene transfer provides a new and efficient means for transfection of various cell types in culture. In this study we evaluate its application to human prostate tumor cells. METHODS. Transient expression of the firefly luciferase gene driven by five viral and five cellular promoters was assessed after in vitro gene transfer using the gene gun method. The relative strengths of these promoters were quantitatively determined in four different human prostate tumor cell lines: DU145, PC-3, LNCaP, and CWR22Rv1 cells. In situ histochemical staining of cells, transfected with bacterial β-galactosidase cDNA as a reporter gene, was also performed to evaluate the transfection efficiency. Time course of gene expression was determined using the luciferase reporter gene. RESULTS. The peak levels of transient expression of firefly luciferase are observed within 24 hr after gene transfer. Sustained but reduced luciferase levels were also detected as long as 5 days post transfection. Up to 35% of bombarded cells in vitro were found to express transgenic β-galactosidase activity. Among tested viral promoters, cytomegalovirus early enhancer/promoter activity was observed to confer consistently the highest activity in each test cell line, whereas phosphoglycerate kinase gene promoter possessed the highest activity among the cellular promoters tested. CONCLUSIONS. The particle bombardment gene-transfer technology can be effectively employed as an efficient method for in vitro gene-transfer into prostate tumor cells. The characterization of relative promoter strength and preference may be useful for future studies of cancer gene therapy approaches.

Original languageEnglish
Pages (from-to)286-292
Number of pages7
JournalProstate
Volume51
Issue number4
DOIs
StatePublished - Jun 1 2002

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Prostatic Neoplasms
Cell Line
Genes
Galactosidases
Transfection
Firefly Luciferases
Prostate
Firearms
Luciferases
Reporter Genes
Phosphoglycerate Kinase
Technology Transfer
Neoplasm Genes
Tumor Cell Line
Cytomegalovirus
Genetic Therapy
Neoplasms
Complementary DNA
Cell Culture Techniques
Staining and Labeling

Keywords

  • Gene gun
  • Gene therapy
  • Promoter
  • Prostate
  • Transfection

ASJC Scopus subject areas

  • Urology

Cite this

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title = "Relative promoter strengths in four human prostate cancer cell lines evaluated by particle bombardment-mediated gene transfer",
abstract = "BACKGROUND. The particle bombardment (gene gun) method for gene transfer provides a new and efficient means for transfection of various cell types in culture. In this study we evaluate its application to human prostate tumor cells. METHODS. Transient expression of the firefly luciferase gene driven by five viral and five cellular promoters was assessed after in vitro gene transfer using the gene gun method. The relative strengths of these promoters were quantitatively determined in four different human prostate tumor cell lines: DU145, PC-3, LNCaP, and CWR22Rv1 cells. In situ histochemical staining of cells, transfected with bacterial β-galactosidase cDNA as a reporter gene, was also performed to evaluate the transfection efficiency. Time course of gene expression was determined using the luciferase reporter gene. RESULTS. The peak levels of transient expression of firefly luciferase are observed within 24 hr after gene transfer. Sustained but reduced luciferase levels were also detected as long as 5 days post transfection. Up to 35{\%} of bombarded cells in vitro were found to express transgenic β-galactosidase activity. Among tested viral promoters, cytomegalovirus early enhancer/promoter activity was observed to confer consistently the highest activity in each test cell line, whereas phosphoglycerate kinase gene promoter possessed the highest activity among the cellular promoters tested. CONCLUSIONS. The particle bombardment gene-transfer technology can be effectively employed as an efficient method for in vitro gene-transfer into prostate tumor cells. The characterization of relative promoter strength and preference may be useful for future studies of cancer gene therapy approaches.",
keywords = "Gene gun, Gene therapy, Promoter, Prostate, Transfection",
author = "Shaobo Zhang and Jian Gu and Yang, {Ning Sun} and Chinghai Kao and Thomas Gardner and John Eble and Liang Cheng",
year = "2002",
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pages = "286--292",
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TY - JOUR

T1 - Relative promoter strengths in four human prostate cancer cell lines evaluated by particle bombardment-mediated gene transfer

AU - Zhang, Shaobo

AU - Gu, Jian

AU - Yang, Ning Sun

AU - Kao, Chinghai

AU - Gardner, Thomas

AU - Eble, John

AU - Cheng, Liang

PY - 2002/6/1

Y1 - 2002/6/1

N2 - BACKGROUND. The particle bombardment (gene gun) method for gene transfer provides a new and efficient means for transfection of various cell types in culture. In this study we evaluate its application to human prostate tumor cells. METHODS. Transient expression of the firefly luciferase gene driven by five viral and five cellular promoters was assessed after in vitro gene transfer using the gene gun method. The relative strengths of these promoters were quantitatively determined in four different human prostate tumor cell lines: DU145, PC-3, LNCaP, and CWR22Rv1 cells. In situ histochemical staining of cells, transfected with bacterial β-galactosidase cDNA as a reporter gene, was also performed to evaluate the transfection efficiency. Time course of gene expression was determined using the luciferase reporter gene. RESULTS. The peak levels of transient expression of firefly luciferase are observed within 24 hr after gene transfer. Sustained but reduced luciferase levels were also detected as long as 5 days post transfection. Up to 35% of bombarded cells in vitro were found to express transgenic β-galactosidase activity. Among tested viral promoters, cytomegalovirus early enhancer/promoter activity was observed to confer consistently the highest activity in each test cell line, whereas phosphoglycerate kinase gene promoter possessed the highest activity among the cellular promoters tested. CONCLUSIONS. The particle bombardment gene-transfer technology can be effectively employed as an efficient method for in vitro gene-transfer into prostate tumor cells. The characterization of relative promoter strength and preference may be useful for future studies of cancer gene therapy approaches.

AB - BACKGROUND. The particle bombardment (gene gun) method for gene transfer provides a new and efficient means for transfection of various cell types in culture. In this study we evaluate its application to human prostate tumor cells. METHODS. Transient expression of the firefly luciferase gene driven by five viral and five cellular promoters was assessed after in vitro gene transfer using the gene gun method. The relative strengths of these promoters were quantitatively determined in four different human prostate tumor cell lines: DU145, PC-3, LNCaP, and CWR22Rv1 cells. In situ histochemical staining of cells, transfected with bacterial β-galactosidase cDNA as a reporter gene, was also performed to evaluate the transfection efficiency. Time course of gene expression was determined using the luciferase reporter gene. RESULTS. The peak levels of transient expression of firefly luciferase are observed within 24 hr after gene transfer. Sustained but reduced luciferase levels were also detected as long as 5 days post transfection. Up to 35% of bombarded cells in vitro were found to express transgenic β-galactosidase activity. Among tested viral promoters, cytomegalovirus early enhancer/promoter activity was observed to confer consistently the highest activity in each test cell line, whereas phosphoglycerate kinase gene promoter possessed the highest activity among the cellular promoters tested. CONCLUSIONS. The particle bombardment gene-transfer technology can be effectively employed as an efficient method for in vitro gene-transfer into prostate tumor cells. The characterization of relative promoter strength and preference may be useful for future studies of cancer gene therapy approaches.

KW - Gene gun

KW - Gene therapy

KW - Promoter

KW - Prostate

KW - Transfection

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SN - 0270-4137

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