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. 1996 May;122(5):275–282. doi: 10.1007/BF01261403

Vincristine induction of mutant and wild-type human multidrug-resistance promoters is cell-type-specific and dose-dependent

Ulrike Stein 1,2, Wolfgang Walther 1, Robert H Shoemaker 1
PMCID: PMC12200739  PMID: 8609150

Abstract

To investigate multidrug-resistance gene (MDR1) promoter efficacy and drug inducibility in cells with different multidrug-resistance phenotypes, multidrug-resistant HCT15 and drug-sensitive KM12 human colon carcinoma cell lines were transfected with constructs incorporating the chloramphenicol acetyltransferase (CAT) reporter gene, driven by wild-type and point-mutatedMDR1 promoter regions. The basalCAT expression level in HCT15 cells was markedly elevated compared to KM12 cells.CAT induction by vincristine was dose-dependent over a broad concentration range (40–500 ng/ml) in both lines. The induction levels were related to the cells' MDR phenotype, with the multidrug-resistant HCT15 cells showing the greater effect. In both cell types, basal and drug-inducedCAT expression were significantly enhanced by the point-mutated promoter regions. The findings support the possible exploitation of theMDR1 promoter for construction of drug-inducible and MDR-cell-targeted expression vectors for use in gene therapy.

Key words: Multidrug resistance, MDR1 promoter, Drug inducibility, CAT assay colon carcinoma cell lines

Abbreviations

MDR1

gene multidrug resistance 1 gene

MDR

multidrug resistance

CAT

chloramphenicol acetyltransferase

RT-PCR

reverse transcriptase/polymerase chain reaction

PBS

phosphate-buffered saline

Footnotes

This work was supported by Feodor-Lynen Fellowships (to U.S. and W.W.) of the Alexander von Humboldt Foundation (Bonn, Germany) and the Office of International Affairs, National Cancer Institute (Bethesda, Md., USA).

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