Attenuation of O(6)-methylguanine-DNA methyltransferase activity and mRNA levels by cisplatin and temozolomide in jurkat cells.
The DNA repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) is important in cellular resistance to certain alkylating antitumor agents such as the methylating drug temozolomide (TMZ). To provide a more rational basis for clinical combinations with another commonly used drug, cisplatin, we assessed the modulation of MGMT protein and mRNA levels in the human leukemic cell line Jurkat after treatment with these agents. Cisplatin decreased MGMT activity in a time- and dose-dependent manner, with maximal suppression (50%) observed 24 h after treatment with 25 microM cisplatin. This was probably the result of decreased transcription of the MGMT gene, because there was an earlier nadir of MGMT mRNA levels after cisplatin treatment and neither cisplatin nor DNA reacted with cisplatin in vitro was able to inhibit MGMT activity in an in vitro assay. TMZ alone depleted MGMT activity in a time- and dose-dependent manner with almost complete loss of activity occurring immediately after treatment with 500 microM TMZ. Combinations of cisplatin (12.5 microM) and TMZ (250 microM) caused substantial and prolonged MGMT depletion with recovery to only 30% of pretreatment levels by 48 h. These results suggest that the clinical efficacy of TMZ and cisplatin may be improved by appropriate schedules of combinations of these agents.
D'Atri
S
S
Istituto Dermopatico Dell'Immacolata, Rome, Italy. s.datri@idi.it
Graziani
G
G
Lacal
P M
PM
Nisticò
V
V
Gilberti
S
S
Faraoni
I
I
Watson
A J
AJ
Bonmassar
E
E
Margison
G P
GP
D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology; D018906:Antineoplastic Agents, Alkylating / Q000008:administration & dosage / Q000494:pharmacology; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D020137:Base Pair Mismatch; D002945:Cisplatin / Q000008:administration & dosage / Q000494:pharmacology*; D004260:DNA Repair; D003606:Dacarbazine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D015870:Gene Expression; D006801:Humans; D019169:Jurkat Cells / Q000187:drug effects* / Q000201:enzymology; D007700:Kinetics; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D000077204:Temozolomide
D000970:Antineoplastic Agents; D018906:Antineoplastic Agents, Alkylating; D012333:RNA, Messenger; D003606:Dacarbazine; D019853:O(6)-Methylguanine-DNA Methyltransferase; D002945:Cisplatin; D000077204:Temozolomide