Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1995 Jan;15(1):290–297. doi: 10.1128/mcb.15.1.290

An XPG DNA repair defect causing mutagen hypersensitivity in mouse leukemia L1210 cells.

J A Vilpo 1, L M Vilpo 1, D E Szymkowski 1, A O'Donovan 1, R D Wood 1
PMCID: PMC231955  PMID: 7799936

Abstract

One of the most widely used antitumor drugs is cis-diamminedichloroplatinum(II) (cisplatin), and mechanisms of cisplatin resistance have been investigated in numerous model systems. Many studies have used mouse leukemia L1210/0 as a reference wild-type cell line, and cisplatin-resistant subclones have been derived from it. Increased DNA excision repair capacity is thought to play a key role in the acquired cisplatin resistance, and this has influenced development of drugs for clinical trials. We report here that the L1210/0 line is in fact severely deficient in nucleotide excision repair of damaged DNA in vivo and in vitro. L1210/0 cell extracts could be complemented by extracts from repair-defective human xeroderma pigmentosum (XP) or rodent excision repair cross-complementing (ERCC) mutant cells, except for XPG/ERCC5 mutants. Purified XPG protein could restore repair proficiency to L1210/0 extracts. Expression of mouse XPG mRNA was similar in all L1210 lines studied, suggesting a point mutation or small alteration of XPG in L1210/0 cells. The DNA repair capacity of a cisplatin-resistant subline, L1210/DDP10, is similar to that of type culture collection L1210 cells and to those of other normal mammalian cell lines. Nucleotide excision repair of DNA is thus clearly important in the intrinsic cellular defense against cisplatin. However, in contrast to what is generally believed, enhancement of DNA repair above the normal level in these rodent cells does not appear to be a mechanism of acquired resistance to the drug.

Full Text

The Full Text of this article is available as a PDF (394.7 KB).

Selected References

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

  1. Aboussekhra A., Wood R. D. Repair of UV-damaged DNA by mammalian cells and Saccharomyces cerevisiae. Curr Opin Genet Dev. 1994 Apr;4(2):212–220. doi: 10.1016/s0959-437x(05)80047-4. [DOI] [PubMed] [Google Scholar]
  2. Alexander M., Curtis G., Avruch J., Goodman H. M. Insulin regulation of protein biosynthesis in differentiated 3T3 adipocytes. Regulation of glyceraldehyde-3-phosphate dehydrogenase. J Biol Chem. 1985 Oct 5;260(22):11978–11985. [PubMed] [Google Scholar]
  3. Andrews P. A., Howell S. B. Cellular pharmacology of cisplatin: perspectives on mechanisms of acquired resistance. Cancer Cells. 1990 Feb;2(2):35–43. [PubMed] [Google Scholar]
  4. Beer J. Z., Budzicka E., Niepokojczycka E., Rosiek O., Szumiel I., Walicka M. Loss of tumorigenicity with simultaneous changes in radiosensitivity and photosensitivity during in vitro growth of L5178Y murine lymphoma cells. Cancer Res. 1983 Oct;43(10):4736–4742. [PubMed] [Google Scholar]
  5. Biggerstaff M., Szymkowski D. E., Wood R. D. Co-correction of the ERCC1, ERCC4 and xeroderma pigmentosum group F DNA repair defects in vitro. EMBO J. 1993 Sep;12(9):3685–3692. doi: 10.1002/j.1460-2075.1993.tb06043.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burchenal J. H., Kalaher K., O'Toole T., Chisholm J. Lack of cross-resistance between certain platinum coordination compounds in mouse leukemia. Cancer Res. 1977 Sep;37(9):3455–3457. [PubMed] [Google Scholar]
  7. Busch D., Greiner C., Lewis K., Ford R., Adair G., Thompson L. Summary of complementation groups of UV-sensitive CHO cell mutants isolated by large-scale screening. Mutagenesis. 1989 Sep;4(5):349–354. doi: 10.1093/mutage/4.5.349. [DOI] [PubMed] [Google Scholar]
  8. Calsou P., Barret J. M., Cros S., Salles B. DNA excision-repair synthesis is enhanced in a murine leukemia L1210 cell line resistant to cisplatin. Eur J Biochem. 1993 Feb 1;211(3):403–409. doi: 10.1111/j.1432-1033.1993.tb17563.x. [DOI] [PubMed] [Google Scholar]
  9. Calvert H., Judson I., van der Vijgh W. J. Platinum complexes in cancer medicine: pharmacokinetics and pharmacodynamics in relation to toxicity and therapeutic activity. Cancer Surv. 1993;17:189–217. [PubMed] [Google Scholar]
  10. Chu G. Cellular responses to cisplatin. The roles of DNA-binding proteins and DNA repair. J Biol Chem. 1994 Jan 14;269(2):787–790. [PubMed] [Google Scholar]
  11. Cleaver J. E., Cortés F., Lutze L. H., Morgan W. F., Player A. N., Mitchell D. L. Unique DNA repair properties of a xeroderma pigmentosum revertant. Mol Cell Biol. 1987 Sep;7(9):3353–3357. doi: 10.1128/mcb.7.9.3353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Collins A. R. Mutant rodent cell lines sensitive to ultraviolet light, ionizing radiation and cross-linking agents: a comprehensive survey of genetic and biochemical characteristics. Mutat Res. 1993 Jan;293(2):99–118. doi: 10.1016/0921-8777(93)90062-l. [DOI] [PubMed] [Google Scholar]
  13. Coverley D., Kenny M. K., Lane D. P., Wood R. D. A role for the human single-stranded DNA binding protein HSSB/RPA in an early stage of nucleotide excision repair. Nucleic Acids Res. 1992 Aug 11;20(15):3873–3880. doi: 10.1093/nar/20.15.3873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eastman A., Schulte N. Enhanced DNA repair as a mechanism of resistance to cis-diamminedichloroplatinum(II). Biochemistry. 1988 Jun 28;27(13):4730–4734. doi: 10.1021/bi00413a022. [DOI] [PubMed] [Google Scholar]
  15. Farrell N., Kelland L. R., Roberts J. D., Van Beusichem M. Activation of the trans geometry in platinum antitumor complexes: a survey of the cytotoxicity of trans complexes containing planar ligands in murine L1210 and human tumor panels and studies on their mechanism of action. Cancer Res. 1992 Sep 15;52(18):5065–5072. [PubMed] [Google Scholar]
  16. Farrell N., Qu Y., Feng L., Van Houten B. Comparison of chemical reactivity, cytotoxicity, interstrand cross-linking and DNA sequence specificity of bis(platinum) complexes containing monodentate or bidentate coordination spheres with their monomeric analogues. Biochemistry. 1990 Oct 16;29(41):9522–9531. doi: 10.1021/bi00493a005. [DOI] [PubMed] [Google Scholar]
  17. Farrell N., Qu Y., Hacker M. P. Cytotoxicity and antitumor activity of bis(platinum) complexes. A novel class of platinum complexes active in cell lines resistant to both cisplatin and 1,2-diaminocyclohexane complexes. J Med Chem. 1990 Aug;33(8):2179–2184. doi: 10.1021/jm00170a021. [DOI] [PubMed] [Google Scholar]
  18. Gibbons G. R., Kaufmann W. K., Chaney S. G. Role of DNA replication in carrier-ligand-specific resistance to platinum compounds in L1210 cells. Carcinogenesis. 1991 Dec;12(12):2253–2257. doi: 10.1093/carcin/12.12.2253. [DOI] [PubMed] [Google Scholar]
  19. Gibbons G. R., Page J. D., Mauldin S. K., Husain I., Chaney S. G. Role of carrier ligand in platinum resistance in L1210 cells. Cancer Res. 1990 Oct 15;50(20):6497–6501. [PubMed] [Google Scholar]
  20. Hansson J., Grossman L., Lindahl T., Wood R. D. Complementation of the xeroderma pigmentosum DNA repair synthesis defect with Escherichia coli UvrABC proteins in a cell-free system. Nucleic Acids Res. 1990 Jan 11;18(1):35–40. doi: 10.1093/nar/18.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hansson J., Keyse S. M., Lindahl T., Wood R. D. DNA excision repair in cell extracts from human cell lines exhibiting hypersensitivity to DNA-damaging agents. Cancer Res. 1991 Jul 1;51(13):3384–3390. [PubMed] [Google Scholar]
  22. Hansson J., Munn M., Rupp W. D., Kahn R., Wood R. D. Localization of DNA repair synthesis by human cell extracts to a short region at the site of a lesion. J Biol Chem. 1989 Dec 25;264(36):21788–21792. [PubMed] [Google Scholar]
  23. Hansson J., Wood R. D. Repair synthesis by human cell extracts in DNA damaged by cis- and trans-diamminedichloroplatinum(II). Nucleic Acids Res. 1989 Oct 25;17(20):8073–8091. doi: 10.1093/nar/17.20.8073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hogan B., Fellous M., Avner P., Jacob F. Isolation of a human teratoma cell line which expresses F9 antigen. Nature. 1977 Dec 8;270(5637):515–518. doi: 10.1038/270515a0. [DOI] [PubMed] [Google Scholar]
  25. Hoy C. A., Thompson L. H., Mooney C. L., Salazar E. P. Defective DNA cross-link removal in Chinese hamster cell mutants hypersensitive to bifunctional alkylating agents. Cancer Res. 1985 Apr;45(4):1737–1743. [PubMed] [Google Scholar]
  26. Jennerwein M. M., Eastman A., Khokhar A. R. The role of DNA repair in resistance of L1210 cells to isomeric 1,2-diaminocyclohexaneplatinum complexes and ultraviolet irradiation. Mutat Res. 1991 Jan;254(1):89–96. doi: 10.1016/0921-8777(91)90044-p. [DOI] [PubMed] [Google Scholar]
  27. Kelland L. R., Mistry P., Abel G., Loh S. Y., O'Neill C. F., Murrer B. A., Harrap K. R. Mechanism-related circumvention of acquired cis-diamminedichloroplatinum(II) resistance using two pairs of human ovarian carcinoma cell lines by ammine/amine platinum(IV) dicarboxylates. Cancer Res. 1992 Jul 15;52(14):3857–3864. [PubMed] [Google Scholar]
  28. MacInnes M. A., Dickson J. A., Hernandez R. R., Learmonth D., Lin G. Y., Mudgett J. S., Park M. S., Schauer S., Reynolds R. J., Strniste G. F. Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe. Mol Cell Biol. 1993 Oct;13(10):6393–6402. doi: 10.1128/mcb.13.10.6393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Masson N., Hurst H. C., Lee K. A. Identification of proteins that interact with CREB during differentiation of F9 embryonal carcinoma cells. Nucleic Acids Res. 1993 Jun 11;21(11):1163–1169. doi: 10.1093/nar/21.5.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nichols A. F., Schmidt W. J., Chaney S. G., Sancar A. Limitations of the in vitro repair synthesis assay for probing the role of DNA repair in platinum resistance. Chem Biol Interact. 1992 Feb;81(3):223–231. doi: 10.1016/0009-2797(92)90079-z. [DOI] [PubMed] [Google Scholar]
  31. Nouspikel T., Clarkson S. G. Mutations that disable the DNA repair gene XPG in a xeroderma pigmentosum group G patient. Hum Mol Genet. 1994 Jun;3(6):963–967. doi: 10.1093/hmg/3.6.963. [DOI] [PubMed] [Google Scholar]
  32. O'Donovan A., Davies A. A., Moggs J. G., West S. C., Wood R. D. XPG endonuclease makes the 3' incision in human DNA nucleotide excision repair. Nature. 1994 Sep 29;371(6496):432–435. doi: 10.1038/371432a0. [DOI] [PubMed] [Google Scholar]
  33. O'Donovan A., Scherly D., Clarkson S. G., Wood R. D. Isolation of active recombinant XPG protein, a human DNA repair endonuclease. J Biol Chem. 1994 Jun 10;269(23):15965–15968. [PubMed] [Google Scholar]
  34. O'Donovan A., Wood R. D. Identical defects in DNA repair in xeroderma pigmentosum group G and rodent ERCC group 5. Nature. 1993 May 13;363(6425):185–188. doi: 10.1038/363185a0. [DOI] [PubMed] [Google Scholar]
  35. Oliver T., Mead G. Testicular cancer. Curr Opin Oncol. 1993 May;5(3):559–567. doi: 10.1097/00001622-199305000-00018. [DOI] [PubMed] [Google Scholar]
  36. Ozols R. F., Young R. C. Chemotherapy of ovarian cancer. Semin Oncol. 1991 Jun;18(3):222–232. [PubMed] [Google Scholar]
  37. Piechaczyk M., Blanchard J. M., Marty L., Dani C., Panabieres F., El Sabouty S., Fort P., Jeanteur P. Post-transcriptional regulation of glyceraldehyde-3-phosphate-dehydrogenase gene expression in rat tissues. Nucleic Acids Res. 1984 Sep 25;12(18):6951–6963. doi: 10.1093/nar/12.18.6951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Richon V. M., Schulte N., Eastman A. Multiple mechanisms of resistance to cis-diamminedichloroplatinum(II) in murine leukemia L1210 cells. Cancer Res. 1987 Apr 15;47(8):2056–2061. [PubMed] [Google Scholar]
  39. Robins P., Jones C. J., Biggerstaff M., Lindahl T., Wood R. D. Complementation of DNA repair in xeroderma pigmentosum group A cell extracts by a protein with affinity for damaged DNA. EMBO J. 1991 Dec;10(12):3913–3921. doi: 10.1002/j.1460-2075.1991.tb04961.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scherly D., Nouspikel T., Corlet J., Ucla C., Bairoch A., Clarkson S. G. Complementation of the DNA repair defect in xeroderma pigmentosum group G cells by a human cDNA related to yeast RAD2. Nature. 1993 May 13;363(6425):182–185. doi: 10.1038/363182a0. [DOI] [PubMed] [Google Scholar]
  41. Sheibani N., Eastman A. Analysis of various mRNA potentially involved in cisplatin resistance of murine leukemia L1210 cells. Cancer Lett. 1990 Jul 31;52(3):179–185. doi: 10.1016/0304-3835(90)90185-z. [DOI] [PubMed] [Google Scholar]
  42. Sheibani N., Jennerwein M. M., Eastman A. DNA repair in cells sensitive and resistant to cis-diamminedichloroplatinum(II): host cell reactivation of damaged plasmid DNA. Biochemistry. 1989 Apr 4;28(7):3120–3124. doi: 10.1021/bi00433a055. [DOI] [PubMed] [Google Scholar]
  43. Shiomi T., Harada Y., Saito T., Shiomi N., Okuno Y., Yamaizumi M. An ERCC5 gene with homology to yeast RAD2 is involved in group G xeroderma pigmentosum. Mutat Res. 1994 Mar;314(2):167–175. doi: 10.1016/0921-8777(94)90080-9. [DOI] [PubMed] [Google Scholar]
  44. Shivji K. K., Kenny M. K., Wood R. D. Proliferating cell nuclear antigen is required for DNA excision repair. Cell. 1992 Apr 17;69(2):367–374. doi: 10.1016/0092-8674(92)90416-a. [DOI] [PubMed] [Google Scholar]
  45. Szymkowski D. E., Lawrence C. W., Wood R. D. Repair by human cell extracts of single (6-4) and cyclobutane thymine-thymine photoproducts in DNA. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):9823–9827. doi: 10.1073/pnas.90.21.9823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Szymkowski D. E., Yarema K., Essigmann J. M., Lippard S. J., Wood R. D. An intrastrand d(GpG) platinum crosslink in duplex M13 DNA is refractory to repair by human cell extracts. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10772–10776. doi: 10.1073/pnas.89.22.10772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Timmer-Bosscha H., Mulder N. H., de Vries E. G. Modulation of cis-diamminedichloroplatinum(II) resistance: a review. Br J Cancer. 1992 Aug;66(2):227–238. doi: 10.1038/bjc.1992.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weeda G., Hoeijmakers J. H. Genetic analysis of nucleotide excision repair in mammalian cells. Semin Cancer Biol. 1993 Apr;4(2):105–117. [PubMed] [Google Scholar]
  49. Wood R. D., Burki H. J. Repair capability and the cellular age response for killing and mutation induction after UV. Mutat Res. 1982 Aug;95(2-3):505–514. doi: 10.1016/0027-5107(82)90281-0. [DOI] [PubMed] [Google Scholar]
  50. Wood R. D., Robins P., Lindahl T. Complementation of the xeroderma pigmentosum DNA repair defect in cell-free extracts. Cell. 1988 Apr 8;53(1):97–106. doi: 10.1016/0092-8674(88)90491-6. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES