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[Cancer Research 51, 3367-3372, July 1, 1991]
© 1991 American Association for Cancer Research
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Effect of O6-Benzylguanine Analogues on Sensitivity of Human Tumor Cells to the Cytotoxic Effects of Alkylating Agents1

M. Eileen Dolan2, R. Brian Mitchell, Christine Mummert, Robert C. Moschel and Anthony E. Pegg

Division of Hematology-Oncology, the University of Chicago Medical Center, Chicago, Illinois 60637 [M. E. D., R. B. M.]; Departments of Cellular and Molecular Physiology and Pharmacology, Pennsylvania State University College of Medicine, The Milton S. Hershey Medical Center, Hershey, Pennsylvania 17033 [C. M., A. E. P.]; and Carcinogen-Modified Nucleic Acid Chemistry, Chemistry of Carcinogenesis Laboratory, ABL-Basic Research Program, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, Maryland 21702 [R. C. M.]

The effect of O6-benzylguanine, O6-(p-chlorobenzyl)guanine, and O6-(p-methylbenzyl)guanine on the sensitivity of various human tumor cell lines to alkylating agents is evaluated. The sensitivity of human colon tumor cells, HT29, to the chloroethylating agents, 1,3-bis(2-chloroethyl)-1-nitrosourea, 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, 2-chloroethyl(methylsulfonyl) methanesulfonate (clomesone), and chlorozotocin was increased by pretreatment for 2 h with 25 µM of each analogue. O6-Benzylguanine was slightly more effective as a sensitizer in HT29 cells than the p-chlorobenzyl and p-methylbenzyl analogues. However, all analogues sensitized SF767 glioma cells to the cytotoxic effects of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, 1,3-bis(2-chloroethyl)-1-nitrosourea, and clomesone to the same degree. Both cell lines were sensitized to the methylating agents streptozotocin and 5-(3-methyl-1-triazeno)imidazole-4-carboxamide, the active intermediate of 5-(3,3-dimethyl-1-triazenyl)imidazole-4-carboxamide, by pretreatment with 10 µM O6-benzylguanine for 2 h. The number of Raji cells surviving 50 µM clomesone decreased 3-fold upon pretreatment for 2 h with 1 µM O6-benzylguanine. The degree of enhancement was dependent on the amount of alkyltransferase protein present in cell lines. For example, HT29 cells (alkyltransferase activity, 381 fmol/mg protein) exhibited a greater degree of enhancement when treated with O6-benzylguanine than SF767 (77 fmol/mg protein) and M19-MEL melanoma (36 fmol/mg protein) cells. There was no enhancement observed in mer- cell lines, U251 (<2 fmol/mg protein), and BE (3 fmol/mg protein), or with alkylating agents which did not produce a cytotoxic lesion at the O6 position of guanine in DNA such as cisplatin or 4-hydroperoxycyclophosphamide. Our studies suggest that O6-benzylguanine analogues may have utility in mer+ tumors as an adjuvant to a variety of alkylating agents which produce a toxic lesion at the O6 position of guanine.

1 This research was supported by the National Cancer Institute, Department of Health and Human Services, through Grants CA-47228 (M.E.D.), CA-18137 (A.E.P.), and Contract NO1-CO-74101 (R.C.M.) with ABL.

2 To whom requests for reprints should be addressed, at Division of Hematology-Oncology, the University of Chicago Medical Center, 5841 S. Maryland Ave., Box 420, Chicago, IL 60637.

Received 2/11/91. Accepted 4/11/91.




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