• 1. Affolter VK, Moore PF. Localized and disseminated histiocytic sarcoma of dendritic cell origin in dogs. Vet Pathol 2002;39:7483.

  • 2. Moore PF, Affolter VK, Vernau W. Canine hemophagocytic histiocytic sarcoma: a proliferative disorder of CD11d+ macrophages. Vet Pathol 2006;43:632645.

    • Search Google Scholar
    • Export Citation
  • 3. Vail DM, Kravis LD, Cooley AJ, et al. Preclinical trial of doxorubicin entrapped in sterically stabilized liposomes in dogs with spontaneously arising malignant tumors. Cancer Che-mother Pharmacol 1997;39:410416.

    • Search Google Scholar
    • Export Citation
  • 4. Takahashi M, Tomiyasu H, Hotta E, et al. Clinical characteristics and prognostic factors in dogs with histiocytic sarcomas in Japan. J Vet Med Sci 2014;76:661666.

    • Search Google Scholar
    • Export Citation
  • 5. Padgett GA, Madewell BR, Keller ET, et al. Inheritance of histiocytosis in Bernese Mountain Dogs. J Small Anim Pract 1995;36:9398.

  • 6. Rassnick KM, Moore AS, Russell DS, et al. Phase II: open-label trial of single-agent CCNU in dogs with previously untreated histiocytic sarcoma. J Vet Intern Med 2010;24:15281531.

    • Search Google Scholar
    • Export Citation
  • 7. Skorupski KA, Clifford CA, Paoloni MC, et al. CCNU for the treatment of dogs with histiocytic sarcoma. J Vet Intern Med 2007;21:121126.

    • Search Google Scholar
    • Export Citation
  • 8. Levine AJ. p53, the cellular gatekeeper for growth and division. Cell 1997;88:323331.

  • 9. Merkel O, Taylor N, Prutsch N, et al. When the guardian sleeps: reactivation of the p53 pathway in cancer. Mutat Res 2017;773:113.

  • 10. Ozaki T, Nakagawara A. p53: the attractive tumor suppressor in the cancer research field. J Biomed Biotechnol 2011;2011:603925.

  • 11. Oshika Y, Nakamura M, Tokunaga T, et al. Multidrug resistance-associated protein and mutant p53 protein expression in non-small cell lung cancer. Mod Pathol 1998;11:10591063.

    • Search Google Scholar
    • Export Citation
  • 12. Wattel E, Preudhomme C, Hecquet B, et al. p53 mutations are associated with resistance to chemotherapy and short survival in hematologic malignancies. Blood 1994;84:31483157.

    • Search Google Scholar
    • Export Citation
  • 13. Kirpensteijn J, Kik M, Teske E, et al. TP53 gene mutations in canine osteosarcoma. Vet Surg 2008;37:454460.

  • 14. Koshino A, Goto-Koshino Y, Setoguchi A, et al. Mutation of p53 gene and its correlation with the clinical outcome in dogs with lymphoma. J Vet Intern Med 2016;30:223229.

    • Search Google Scholar
    • Export Citation
  • 15. Van Leeuwen IS, Hellmen E, Cornelisse CJ, et al. P53 mutations in mammary tumor cell lines and corresponding tumor tissues in the dog. Anticancer Res 1996;16:37373744.

    • Search Google Scholar
    • Export Citation
  • 16. York D, Higgins RJ, LeCouteur RA, et al. TP53 mutations in canine brain tumors. Vet Pathol 2012;49:796801.

  • 17. Asada H, Tsuboi M, Chambers JK, et al. A 2-base insertion in exon 5 is a common mutation of the TP53 gene in dogs with histiocytic sarcoma. J Vet Med Sci 2017;79:17211726.

    • Search Google Scholar
    • Export Citation
  • 18. Wellman ML, Krakowka S, Jacobs RM, et al. A macrophagemonocyte cell line from a dog with malignant histiocytosis. In Vitro Cell Dev Biol 1988;24:223229.

    • Search Google Scholar
    • Export Citation
  • 19. Azakami D, Bonkobara M, Washizu T, et al. Establishment and biological characterization of canine histiocytic sarcoma cell lines. J Vet Med Sci 2006;68:13431346.

    • Search Google Scholar
    • Export Citation
  • 20. Asada H, Tomiyasu H, Goto-Koshino Y, et al. Evaluation of the drug sensitivity and expression of 16 drug resistance-related genes in canine histiocytic sarcoma cell lines. J Vet Med Sci 2015;77:677684.

    • Search Google Scholar
    • Export Citation
  • 21. Tomiyasu H, Goto-Koshino Y, Takahashi M, et al. Quantitative analysis of mRNA for 10 different drug resistance factors in dogs with lymphoma. J Vet Med Sci 2010;72:11651172.

    • Search Google Scholar
    • Export Citation
  • 22. Rivera-Calderón LG, Fonseca-Alves CE, Kobayashi PE, et al. Alterations in PTEN, MDM2, TP53 and AR protein and gene expression are associated with canine prostate carcinogenesis. Res Vet Sci 2016;106:5661.

    • Search Google Scholar
    • Export Citation
  • 23. Peters IR, Peeters D, Helps CR, et al. Development and application of multiple internal reference (housekeeper) gene assays for accurate normalisation of canine gene expression studies. Vet Immunol Immunopathol 2007;117:5566.

    • Search Google Scholar
    • Export Citation
  • 24. Nasif S, Contu L, Muhlemann O. Beyond quality control: the role of nonsense-mediated mRNA decay (NMD) in regulating gene expression. Semin Cell Dev Biol 2018;75:7887.

    • Search Google Scholar
    • Export Citation
  • 25. Nickless A, Bailis JM, You Z. Control of gene expression through the nonsense-mediated RNA decay pathway. Cell Biosci 2017;7:26.

  • 26. Hedan B, Thomas R, Motsinger-Reif A, et al. Molecular cytogenetic characterization of canine histiocytic sarcoma: a spontaneous model for human histiocytic cancer identifies deletion of tumor suppressor genes and highlights influence of genetic background on tumor behavior. BMC Cancer 2011;11:201.

    • Search Google Scholar
    • Export Citation
  • 27. Zhang J, Chen X, Kent MS, et al. Establishment of a dog model for the p53 family pathway and identification of a novel isoform of p21 cyclin-dependent kinase inhibitor. Mol Cancer Res 2009;7:6778.

    • Search Google Scholar
    • Export Citation
  • 28. Gu J, Hu W, Song ZP, et al. Resveratrol-induced autophagy promotes survival and attenuates doxorubicin-induced cardiotoxicity. Int Immunopharmacol 2016;32:17.

    • Search Google Scholar
    • Export Citation
  • 29. Batista LF, Roos WP, Christmann M, et al. Differential sensitivity of malignant glioma cells to methylating and chloroethylating anticancer drugs: p53 determines the switch by regulating xpc, ddb2, and DNA double-strand breaks. Cancer Res 2007;67:1188611895.

    • Search Google Scholar
    • Export Citation
  • 30. Ikeda J. Roles of p53 in chemotherapy of glioblastoma. Hokkaido Igaku Zasshi 2000;75:299314.

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Effect of a two-base insertion mutation of the TP53 gene on expression of p53 protein in canine histiocytic sarcoma cells

Hajime Asada DVM1, Hirotaka Tomiyasu PhD, DVM1, Yuko Goto-Koshino PhD, DVM1, Koichi Ohno PhD, DVM1, and Hajime Tsujimoto PhD, DVM1
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  • 1 1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113–8657, Japan.

Abstract

OBJECTIVE

To examine effects of a common mutation (2-base insertion in exon 5) of the TP53 gene on biological function of p53 protein in canine histiocytic sarcoma cells.

SAMPLE

Canine histiocytic tumor cell lines DH82 with deletion of TP53 and CHS-3 with the wild-type TP53 and canine wild-type and mutant TP53 fragments.

PROCEDURES

Wild-type or mutant TP53 with a polyprotein peptide tag at the N-terminus was transduced into DH82 and CHS-3 cells. Expression of p53 protein, changes in function as a transcription factor, and susceptibility to doxorubicin and nimustine were compared.

RESULTS

Transduced p53 protein was detected in wild-type TP53–transduced DH82 and CHS-3 cells, whereas expression was not detected in mutant TP53–transduced cells. There were significant increases in expression of target genes of p53 protein, including p21 and MDM2, in wild-type TP53–transduced cells, compared with results for native and mock-transfected cells, but not in mutant TP53–transduced cells. There was no significant difference in drug susceptibilities among native and derivative cells of CHS-3. However, cell viabilities of wild-type TP53–transduced DH82 cells incubated with doxorubicin were significantly lower than viabilities of native, mock-transfected, and AT insertion mutation–TP53–transduced DH82 cells; susceptibility to nimustine did not differ significantly among cells.

CONCLUSIONS AND CLINICAL RELEVANCE

Expression of p53 protein and its function as a transcription factor were lost after addition of a 2-base insertion in the TP53 gene in canine histiocytic tumor cells. Additional studies are needed to investigate the clinical relevance of this mutation in histiocytic sarcomas of dogs.

Abstract

OBJECTIVE

To examine effects of a common mutation (2-base insertion in exon 5) of the TP53 gene on biological function of p53 protein in canine histiocytic sarcoma cells.

SAMPLE

Canine histiocytic tumor cell lines DH82 with deletion of TP53 and CHS-3 with the wild-type TP53 and canine wild-type and mutant TP53 fragments.

PROCEDURES

Wild-type or mutant TP53 with a polyprotein peptide tag at the N-terminus was transduced into DH82 and CHS-3 cells. Expression of p53 protein, changes in function as a transcription factor, and susceptibility to doxorubicin and nimustine were compared.

RESULTS

Transduced p53 protein was detected in wild-type TP53–transduced DH82 and CHS-3 cells, whereas expression was not detected in mutant TP53–transduced cells. There were significant increases in expression of target genes of p53 protein, including p21 and MDM2, in wild-type TP53–transduced cells, compared with results for native and mock-transfected cells, but not in mutant TP53–transduced cells. There was no significant difference in drug susceptibilities among native and derivative cells of CHS-3. However, cell viabilities of wild-type TP53–transduced DH82 cells incubated with doxorubicin were significantly lower than viabilities of native, mock-transfected, and AT insertion mutation–TP53–transduced DH82 cells; susceptibility to nimustine did not differ significantly among cells.

CONCLUSIONS AND CLINICAL RELEVANCE

Expression of p53 protein and its function as a transcription factor were lost after addition of a 2-base insertion in the TP53 gene in canine histiocytic tumor cells. Additional studies are needed to investigate the clinical relevance of this mutation in histiocytic sarcomas of dogs.

Supplementary Materials

    • Supplementary Appendix s1 (PDF 153 kb)
    • Supplementary Figure s1 (PDF 194 kb)

Contributor Notes

Address correspondence to Dr. Tomiyasu (atomi@mail.ecc.u-tokyo.ac.jp).