Enhanced DNA repair and tolerance of DNA damage associated with resistance to cis-diammine-dichloroplatinum (II) after in vitro exposure of a human teratoma cell line to fractionated X-irradiation.
In vitro exposure of a human testicular teratoma continuous cell line to fractionated X-irradiation resulted in the expression of resistance to cisplatin. In two independently-derived sublines, designated SUSA-DXR13 and SUSA-DXR10 resulting from treatment with either 13 fractions of 1.5 Gy (dose required to reduce survival by 1 log) or 10 fractions of 3 Gy (dose required to reduce survival by 2 logs) respectively, the IC50 values for cisplatin were 2- and 3.1-fold higher than that of the parental cell line. These sublines were cross-resistant to carboplatin (approximately 2-fold) but not to adriamycin and they showed unaltered radiosensitivities. The SUSA-DXR10 subline expressed some cross-resistance to mitomycin C and melphalan but none to Carmustine (BCNU). Total glutathione content was significantly reduced in both SUSA-DXR10 and SUSA-DXR13 cells, but the activities of associated enzymes, including the glutathione S-transferases, peroxidase and reductase were not modified significantly in the resistant sublines. Resistance in the SUSA-DXR10 subline was associated with significantly decreased 195mcisplatin uptake (p less than 0.01), but this was not reflected in a reduced level of drug bound to the DNA. The formation and removal of four platinum-DNA adducts were immunochemically quantitated. Immediately following drug treatment there was a higher level of total platination of the DNA in the resistant subline indicative of increased tolerance to DNA damage. After an 18 hr post treatment incubation, there was an indication of some repair capacity in this SUSA-DXR10 cell line, which was not apparent in the parental cells. Neither the parental nor the SUSA-DXR10 cell line was proficient in the repair of the major adduct Pt-GG, whereas both lines repaired the monofunctional adduct and the adduct Pt(GMP)2. SUSA-DXR10 cells were also able to repair the intrastrand adduct Pt-AG and interstrand crosslinks, unlike the repair deficient parental cells. Higher levels of interstrand crosslinks were characteristic of the SUSA-DXR10 subline. These observations therefore implicate both enhanced repair and increased tolerance of DNA damage as mechanisms of resistance to cisplatin resulting from in vitro exposure of a human teratoma cell line to fractionated X-irradiation.
Hill
B T
BT
Laboratory of Cellular Chemotherapy, Imperial Cancer Research Fund, Lincoln's Inn Fields, London, U.K.
Shellard
S A
SA
Hosking
L K
LK
Fichtinger-Schepman
A M
AM
Bedford
P
P
D002945:Cisplatin / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D018736:DNA Adducts; D004249:DNA Damage; D004260:DNA Repair / Q000528:radiation effects*; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000528:radiation effects*; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008297:Male; D011879:Radiotherapy Dosage; D013724:Teratoma / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D013736:Testicular Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000201:enzymology / Q000528:radiation effects*
D018736:DNA Adducts; C057270:cisplatin-DNA adduct; D004247:DNA; D005979:Glutathione Peroxidase; D005980:Glutathione Reductase; D005982:Glutathione Transferase; D005978:Glutathione; D002945:Cisplatin