Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1979 Oct;76(10):5249–5253. doi: 10.1073/pnas.76.10.5249

HeLa cell variants that differ in sensitivity to monofunctional alkylating agents, with independence of cytotoxic and mutagenic responses

R M Baker 1,2, W C Van Voorhis 1,2,*, L A Spencer 1,2
PMCID: PMC413118  PMID: 291942

Abstract

Different strains of the established human cell line HeLa differ substantially in sensitivity to ethyl methanesulfonate (EtMes). The EtMes doses effective for either cytotoxicity or mutation induction in a line of HeLa S3 cells are about 1/10th those required in the CCL2 HeLa line of the American Type Culture Collection. By plating the sensitive HeLa S3 line in the presence of highly cytotoxic doses of EtMes, we obtained a clone (designated A6) that displays about 7-fold greater resistance to EtMes toxicity. This A6 isolate is also cross resistant to other simple monofunctional alkylating agents—exhibiting about 4-fold increased resistance to methyl methanesulfonate and 10- to 15-fold increased resistance to N-methyl-N′-nitro-N-nitrosoguanidine but is similar to the S3 parent in sensitivity to mitomycin C, UV radiation, and γ-rays. In contrast to the results for cytotoxicity, the A6 variant and the S3 parent showed the same high susceptibility to EtMes induction of ouabain-resistant mutations. This is direct biological evidence that different alkylation lesions are normally responsible for mutagenic and cytotoxic effects. The S3 and A6 cell lines may differ in DNA repair capability specific to certain potentially lethal alkylation products. The comparative sensitivity of the A6 cells to alkylation mutagenesis may also prove useful in cell genetic studies by facilitating the generation of multiple mutants for recessive alleles and permitting exceptionally sensitive detection of specific mutagenic effects.

Keywords: somatic cell mutants, ethyl methanesulfonate, ouabain resistance, DNA repair

Full text

PDF
5252

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boyd J. B., Golino M. D., Nguyen T. D., Green M. M. Isolation and characterization of X-linked mutants of Drosophila melanogaster which are sensitive to mutagens. Genetics. 1976 Nov;84(3):485–506. doi: 10.1093/genetics/84.3.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brendel M., Haynes R. H. Interactions among genes controlling sensitivity to radiation and alkylation in yeast. Mol Gen Genet. 1973 Sep 12;125(3):197–216. doi: 10.1007/BF00270743. [DOI] [PubMed] [Google Scholar]
  3. Buchwald M. Mutagenesis at the ouabain-resistance locus in human diploid fibroblasts. Mutat Res. 1977 Sep;44(3):401–411. doi: 10.1016/0027-5107(77)90098-7. [DOI] [PubMed] [Google Scholar]
  4. Fujiwara Y., Tatsumi M. Cross-link repair in human cells and its possible defect in Fanconi's anemia cells. J Mol Biol. 1977 Jul 15;113(4):635–649. doi: 10.1016/0022-2836(77)90227-3. [DOI] [PubMed] [Google Scholar]
  5. HAYNES R. H., INCH W. R. SYNERGISTIC ACTION OF NITROGEN MUSTARD AND RADIATION IN MICROORGANISMS. Proc Natl Acad Sci U S A. 1963 Nov;50:839–846. doi: 10.1073/pnas.50.5.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Higgins N. P., Strauss B. S. Differences in the ability of human lymphoblastoid lines to exclude bromodeoxyuridine and in their sensitivity to methyl methanesulfonate and to incorporated [3H]thymidine. Cancer Res. 1979 Feb;39(2 Pt 1):312–320. [PubMed] [Google Scholar]
  7. Hoar D. I., Sargent P. Chemical mutagen hypersensitivity in ataxia telangiectasia. Nature. 1976 Jun 17;261(5561):590–592. doi: 10.1038/261590a0. [DOI] [PubMed] [Google Scholar]
  8. Jeggo P., Defais M., Samson L., Schendel P. The adaptive response of E.coli to low levels of alkylating agent: the role of polA in killing adaptation. Mol Gen Genet. 1978 Jul 4;162(3):299–305. doi: 10.1007/BF00268855. [DOI] [PubMed] [Google Scholar]
  9. Kao F. T., Puck T. T. Genetics of somatic mammalian cells. IX. Quantitation of mutagenesis by physical and chemical agents. J Cell Physiol. 1969 Dec;74(3):245–258. doi: 10.1002/jcp.1040740305. [DOI] [PubMed] [Google Scholar]
  10. Lawley P. D., Orr D. J. Specific excision of methylation products from DNA of Escherichia coli treated with N-methyl-N'-nitro-N-nitrosoguanidine. Chem Biol Interact. 1970 Aug;2(2):154–157. doi: 10.1016/0009-2797(70)90047-5. [DOI] [PubMed] [Google Scholar]
  11. Lawley P. D. Some chemical aspects of dose-response relationships in alkylation mutagenesis. Mutat Res. 1974 Jun;23(3):283–295. doi: 10.1016/0027-5107(74)90102-x. [DOI] [PubMed] [Google Scholar]
  12. Lindahl T. DNA glycosylases, endonucleases for apurinic/apyrimidinic sites, and base excision-repair. Prog Nucleic Acid Res Mol Biol. 1979;22:135–192. doi: 10.1016/s0079-6603(08)60800-4. [DOI] [PubMed] [Google Scholar]
  13. Ljungquist S., Lindahl T., Howard-Flanders P. Methyl methane sulfonate-sensitive mutant of Escherichia coli deficient in an endonuclease specific for apurinic sites in deoxyribonucleic acid. J Bacteriol. 1976 May;126(2):646–653. doi: 10.1128/jb.126.2.646-653.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mankovitz R., Buchwald M., Baker R. M. Isolation of ouabain-resistant human diploid fibroblasts. Cell. 1974 Nov;3(3):221–226. doi: 10.1016/0092-8674(74)90135-4. [DOI] [PubMed] [Google Scholar]
  15. PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pegg A. E. Formation and metabolism of alkylated nucleosides: possible role in carcinogenesis by nitroso compounds and alkylating agents. Adv Cancer Res. 1977;25:195–269. doi: 10.1016/s0065-230x(08)60635-1. [DOI] [PubMed] [Google Scholar]
  17. Peterson A. R., Peterson H., Heidelberger C. Oncogenesis, mutagenesis, DNA damage, and cytotoxicity in cultured mammalian cells treated with alkylating agents. Cancer Res. 1979 Jan;39(1):131–138. [PubMed] [Google Scholar]
  18. Prakash L., Prakash S. Isolation and characterization of MMS-sensitive mutants of Saccharomyces cerevisiae. Genetics. 1977 May;86(1):33–55. doi: 10.1093/genetics/86.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Robbins A. R., Baker R. M. (Na, K)ATPase activity in membrane preparations of ouabain-resistant HeLa cells. Biochemistry. 1977 Nov 15;16(23):5163–5168. doi: 10.1021/bi00642a600. [DOI] [PubMed] [Google Scholar]
  20. Roberts J. J., Pascoe J. M., Plant J. E., Sturrock J. E., Crathorn A. R. Quantitative aspects of the repair of alkylated DNA in cultured mammalian cells. I. The effect on HeLa and Chinese hamster cell survival of alkylation of cellular macromolecules. Chem Biol Interact. 1971 Feb;3(1):29–47. doi: 10.1016/0009-2797(71)90024-x. [DOI] [PubMed] [Google Scholar]
  21. Schendel P. F., Defais M., Jeggo P., Samson L., Cairns J. Pathways of mutagenesis and repair in Escherichia coli exposed to low levels of simple alkylating agents. J Bacteriol. 1978 Aug;135(2):466–475. doi: 10.1128/jb.135.2.466-475.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schendel P. F., Robins P. E. Repair of O6-methylguanine in adapted Escherichia coli. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6017–6020. doi: 10.1073/pnas.75.12.6017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stich H. F., San R. H., Kawazoe Y. Increased sensitivity of xeroderma pigmentosum cells to some chemical carcinogens and mutagens. Mutat Res. 1973 Jan;17(1):127–137. doi: 10.1016/0027-5107(73)90261-3. [DOI] [PubMed] [Google Scholar]
  24. Strauss B. S. DNA repair mechanisms and their relation to mutation and recombination. Curr Top Microbiol Immunol. 1968;44:1–85. [PubMed] [Google Scholar]
  25. Taylor A. M., Harnden D. G., Arlett C. F., Harcourt S. A., Lehmann A. R., Stevens S., Bridges B. A. Ataxia telangiectasia: a human mutation with abnormal radiation sensitivity. Nature. 1975 Dec 4;258(5534):427–429. doi: 10.1038/258427a0. [DOI] [PubMed] [Google Scholar]
  26. Thompson L. H., Baker R. M. Isolation of mutants of cultured mammalian cells. Methods Cell Biol. 1973;6:209–281. doi: 10.1016/s0091-679x(08)60052-7. [DOI] [PubMed] [Google Scholar]
  27. Yamamoto Y., Katsuki M., Sekiguchi M., Otsuji N. Escherichia coli gene that controls sensitivity to alkylating agents. J Bacteriol. 1978 Jul;135(1):144–152. doi: 10.1128/jb.135.1.144-152.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES