Editorial Type:
Oncology

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

Hajime Asada 1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113–8657, Japan.

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Hirotaka Tomiyasu 1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113–8657, Japan.

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Yuko Goto-Koshino 1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113–8657, Japan.

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Koichi Ohno 1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113–8657, Japan.

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Hajime Tsujimoto 1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113–8657, Japan.

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DOI:
https://doi.org/10.2460/ajvr.80.7.680
Volume/Issue:
Volume 80: Issue 7
Online Publication Date:
01 Jul 2019
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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).
Cover American Journal of Veterinary Research
American Journal of Veterinary Research
Publication Date:
01 Jul 2019

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