Asada Hajime, Tomiyasu Hirotaka, Goto-Koshino Yuko, Ohno Koichi, Tsujimoto Hajime
Am J Vet Res. 2019 Jul;80(7):680-688. doi: 10.2460/ajvr.80.7.680.
To examine effects of a common mutation (2-base insertion in exon 5) of the gene on biological function of p53 protein in canine histiocytic sarcoma cells.
Canine histiocytic tumor cell lines DH82 with deletion of and CHS-3 with the wild-type and canine wild-type and mutant fragments.
Wild-type or mutant 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.
Transduced p53 protein was detected in wild-type -transduced DH82 and CHS-3 cells, whereas expression was not detected in mutant -transduced cells. There were significant increases in expression of target genes of p53 protein, including and , in wild-type -transduced cells, compared with results for native and mock-transfected cells, but not in mutant -transduced cells. There was no significant difference in drug susceptibilities among native and derivative cells of CHS-3. However, cell viabilities of wild-type -transduced DH82 cells incubated with doxorubicin were significantly lower than viabilities of native, mock-transfected, and AT insertion mutation--transduced DH82 cells; susceptibility to nimustine did not differ significantly among cells.
Expression of p53 protein and its function as a transcription factor were lost after addition of a 2-base insertion in the gene in canine histiocytic tumor cells. Additional studies are needed to investigate the clinical relevance of this mutation in histiocytic sarcomas of dogs.
研究基因的一种常见突变（外显子5中2个碱基插入）对犬组织细胞肉瘤细胞中p53蛋白生物学功能的影响。
缺失该基因的犬组织细胞瘤细胞系DH82、野生型该基因的CHS - 3以及犬野生型和突变型该基因片段。
将N端带有多聚蛋白肽标签的野生型或突变型该基因转导至DH82和CHS - 3细胞中。比较p53蛋白的表达、作为转录因子的功能变化以及对阿霉素和尼莫司汀的敏感性。
在野生型该基因转导的DH82和CHS - 3细胞中检测到转导的p53蛋白，而在突变型该基因转导的细胞中未检测到表达。与天然细胞和模拟转染细胞相比，野生型该基因转导的细胞中p53蛋白靶基因（包括和）的表达显著增加，但突变型该基因转导的细胞中未增加。CHS - 3的天然细胞和衍生细胞之间的药物敏感性无显著差异。然而，用阿霉素处理的野生型该基因转导的DH82细胞的细胞活力显著低于天然、模拟转染以及AT插入突变转导的DH82细胞；细胞对尼莫司汀的敏感性在各细胞间无显著差异。
在犬组织细胞瘤细胞的该基因中添加2个碱基插入后，p53蛋白的表达及其作为转录因子的功能丧失。需要进一步研究来探讨这种突变在犬组织细胞肉瘤中的临床意义。
