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The expression of topoisomerase II alpha mRNA is increased by the minor groove DNA‐binding drug Hoechst 33258 in African green monkey kidney (Vero) cells
Author(s) -
Thomsen Kristen M.,
Daggett Melissa A. F.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a78-b
Subject(s) - topoisomerase , microbiology and biotechnology , transfection , biology , vero cell , dna , gene , luciferase , cell culture , messenger rna , gene expression , cancer cell , cancer , genetics
Topoisomerase IIα is a gene that plays a critical role in the replication of DNA. Several anti‐tumor chemotherapeutic agents selectively target the topoisomerase IIαprotein and promote the death of rapidly dividing tumor cells. In many drug resistant tumors, topoisomerase IIαhas been shown to be transcriptionally down regulated. This lowered level of topoisomerase IIαexpression has an adverse effect on the chemosensitivity of tumors to anti‐tumor chemotherapeutic agents. In this study, we examined the effect of the DNA minor groove‐binding agent Hoechst 33258 on the expression of topoisomerase IIα mRNA in subconfluent Vero and MCF‐7 cell lines. While we did not detect endogenous expression of topoisomerase IIα mRNA in the MCF‐7 cells, the addition of hoechst 33258 to Vero cells resulted in increased expression of topoisomerase IIα mRNA. In order to further investigate these results, we have amplified and cloned the promoter regions of topoisomerase IIα from both Vero and human genomic DNA into the luciferase reporter vector pGL3. Transient transfection analysis and sequence comparisons of the promoter regions from both human and monkey genomic DNA will provide additional information as to the role conserved promoter elements have in promoting the up regulation of topoisomerase IIα in response to Hoechst 33258. Results from these studies will help to identify sequence specific promoter elements that can be targeted for modifying the expression of genes known to be influential in the chemosensitivity of cancer cells. This research was supported through funding from NIH Grant 1R15CA096723‐01