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[Cancer Research 56, 5375-5379, December 1, 1996]
© 1996 American Association for Cancer Research
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Mismatch Repair Mutations Override Alkyltransferase in Conferring Resistance to Temozolomide but not to 1,3-Bis(2-chloroethyl)nitrosourea1

Lili Liu, Sanford Markowitz and Stanton L. Gerson2

Division of Hematology and Oncology, Department of Medicine, and the University/Ireland Cancer Research Center, Case Western Reserve University School of Medicine, and University Hospital of Cleveland, Cleveland, Ohio 44106-4937

Cells with the mutator phenotype are tolerant to methylating damage from N-methylnitrosourea and N-methyl-N'-nitro-N-nitrosoguanine, exhibit replication repair errors, and have recently been found to be mutant in mismatch repair (MMR). However, resistance of cell lines with these defects to clinically used chemotherapeutic agents and the relationship of this resistance to expression of O6-alkylguanine-DNA alkyltransferase (AGT), which repairs DNA damage caused by methylating agents, has not been demonstrated. We compared resistance to the methylating agent temozolomide (TMZ) and to the chloroethylating agent 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), with and without AGT inhibition by O6-bG in several colorectal carcinoma cell lines. Two cell lines had known microsatellite instability (replication repair error-positive) and high levels of AGT, as well as a mutation in one of two MMR genes, hMLH1 (HCT116) or GTBP (HCT15). Cell line SW480 had wild-type MMR genes and high AGT, and HCT116+Ch3 has previously been transduced with chromosome 3 (carrying wild-type hMLH1) and thus has a "corrected" MMR phenotype. SW480 exhibited the expected sensitivity to TMZ and BCNU and marked potentiation of cytotoxicity by O6-bG. In contrast, HCT15 and HCT116 were markedly resistant to TMZ and were not sensitized by O6-bG-mediated inhibition of AGT, whereas the sensitivity pattern in HCT116+Ch3 cells was similar to that in SW480. All cell lines were sensitized to BCNU by O6-bG. Thus, tumor cells with defects in MMR appear particularly resistant to methylating agents in a manner that overrides dependence on AGT and its inhibition by O6-bG. However, these cells use AGT for resistance to chloroethylating agents, providing an alternative strategy for alkylating agent therapy.

1 Supported in part by the following grants from the Public Health Service: R01ES06288, UO1CA75525, R01CA63193 and P30CA43703.

2 To whom requests for reprints should be addressed, at Division of Hematology and Oncology, Case Western Reserve University School of Medicine, BRB-3, 10900 Euclid Avenue, Cleveland, OH 44106-4937.

Received 8/ 7/96. Accepted 10/17/96.




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