Preferential platination of an activated cellular promoter by cis-diamminedichloroplatinum.
This study examines how accessibility to cisplatin on various genomic regions in T47D breast cancer cells, including the retinoic acid receptor beta gene promoter and coding region and the dihydrofolate reductase gene promoter and coding region, is affected by treatment of the cells with 9-cis retinoic acid, a treatment that activates the retinoic acid receptor beta gene promoter in these cells. A PCR-based assay was used to measure cisplatin adduct density based on the inhibition of PCR amplification of templates from cisplatin treated versus untreated cells. Treatment of cells with 9-cis retinoic acid enhanced accessibility to cisplatin on the retinoic acid receptor beta gene promoter region, but not on the coding regions of that gene nor on the dihydrofolate reductase gene promoter or coding regions, where accessibilities to cisplatin remained 2-4 times lower than on the activated retinoic acid receptor beta gene promoter. Examination of smaller regions within this promoter region showed a repression of platination in the 500 bp region surrounding the TATA box in cells prior to 9-cis retinoic acid treatment, which was abolished following promoter activation. Differences in sequence composition between the various regions could not fully account for differences in platination, suggesting that structural features such as bends in retinoic acid receptor beta gene promoter DNA following gene activation, create energetically favorable sites for platination, and contribute to the cytotoxicity of the drug.
Haghighi
A
A
Sidney Kimmel Cancer Center, San Diego, California 92121, USA.
Lebedeva
S
S
Gjerset
R A
RA
D000077556:Alitretinoin; D000970:Antineoplastic Agents / Q000494:pharmacology; D001943:Breast Neoplasms; D002945:Cisplatin / Q000378:metabolism* / Q000494:pharmacology*; D018736:DNA Adducts / Q000378:metabolism; D004273:DNA, Neoplasm / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D015533:Transcriptional Activation; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D018736:DNA Adducts; D004273:DNA, Neoplasm; D018168:Receptors, Retinoic Acid; C082925:retinoic acid receptor beta; D000077556:Alitretinoin; D014212:Tretinoin; D013762:Tetrahydrofolate Dehydrogenase; D002945:Cisplatin