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Relative promoter strengths in four human prostate cancer cell lines evaluated by particle bombardment‐mediated gene transfer *
Author(s) -
Zhang Shaobo,
Gu Jian,
Yang NingSun,
Kao Chinghai,
Gardner Thomas A.,
Eble John N.,
Cheng Liang
Publication year - 2002
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/pros.10081
Subject(s) - transfection , luciferase , promoter , microbiology and biotechnology , reporter gene , biology , lncap , cell culture , gene expression , gene , cancer cell , cancer , biochemistry , genetics
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. Prostate 51: 286–292, 2002. © 2002 Wiley‐Liss, Inc.