Activated ras oncogene and specifically acquired resistance to cisplatin in human mammary epithelial cells: induction of DNA cross-links and their repair.
A human non-malignant mammary epithelial cell line, HBL100, and the ras-transformed HBL100/ras1 cell line were examined for their sensitivity to cis-diamminedichloroplatinum(II) (cisplatin). The clonogenic cell survival assay showed that HBL100/ras1 exhibited a 2.7-fold increased resistance compared to the parental HBL100 cell line. The responses to other agents interacting with DNA, such as mitomycin C, 8-methoxypsoralen plus UVA or doxorubicin, were very similar in both cell lines. The same is true for ionizing radiation (Alapetite et al., Int J. Radiat. Biol., 59, 385-396, 1991). In other words, the mechanism of acquired resistance in HBL100 appears to be limited to cisplatin. No difference was observed between the two cell lines in cisplatin uptake as determined by atomic absorption spectrometry. Alkaline elution showed that less interstrand cross-links were formed by this drug in the resistant HBL100/ras1 cells compared to HBL100 and, moreover, the removal of these adducts was clearly more efficient in the former cell line. This was confirmed by an in vitro excision repair assay which revealed a 2.2-fold increase in DNA repair activity in the extracts from HBL100/ras1 versus HBL100 cells. It is concluded that the transformation of human epithelial HBL100 cells by the ras gene resulted in an acquired resistance apparently limited to cisplatin, a feature associated with a reduced proportion of induced interstrand cross-links and a higher efficiency in their removal. The mechanism of involvement of the ras gene product in this process is still a matter of speculation.
Levy
E
E
Institut Curie, Section de Biologie, URA 1292 du CNRS, Paris, France.
Baroche
C
C
Barret
J M
JM
Alapetite
C
C
Salles
B
B
Averbeck
D
D
Moustacchi
E
E
D001940:Breast / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002945:Cisplatin / Q000493:pharmacokinetics / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004317:Doxorubicin / Q000494:pharmacology; D004351:Drug Resistance; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005260:Female; D005786:Gene Expression Regulation; D011905:Genes, ras; D006801:Humans; D008730:Methoxsalen / Q000494:pharmacology; D016685:Mitomycin / Q000494:pharmacology; D012680:Sensitivity and Specificity
D016685:Mitomycin; D004317:Doxorubicin; D004247:DNA; D002945:Cisplatin; D008730:Methoxsalen