Conrad David, Kehl Alexandra, Beitzinger Christoph, Metzler Thomas, Steiger Katja, Pfarr Nicole, Fischer Konrad, Klopfleisch Robert, Aupperle-Lellbach Heike
Department of Pathology, LABOKLIN GmbH & Co. KG, 97688 Bad Kissingen, Germany.
Department of Molecular Biology, LABOKLIN GmbH & Co. KG, 97688 Bad Kissingen, Germany.
Vet Sci. 2022 Jan 30;9(2):56. doi: 10.3390/vetsci9020056.
Canine digital melanoma, in contrast to canine oral melanoma, is still largely unexplored at the molecular genetic level. The aim of this study was to detect mutant genes in digital melanoma. Paraffin-embedded samples from 86 canine digital melanomas were examined for the V595E variant by digital droplet PCR (ddPCR), and for exon 11 mutations in Furthermore, exons 2 and 3 of and were analysed by Sanger sequencing. Copy number variations (CNV) of in genomic DNA were analysed from nine dogs. The V595E variant was absent and in , a single nucleotide polymorphism was found in 16/70 tumours (23%). The number of copies of varied between 4 and 6. exon 2 codons 12 and 13 were mutated in 22/86 (25.6%) of the melanomas examined. Other mutually exclusive mutations were found within the hotspot loci, i.e., exon 3 codon 61: 2/55 (3.6%); exon 2 codons 12 and 13: 2/83 (2.4%); and exon 3 codon 61: 9/86 (10.5%). However, no correlation could be established between histological malignancy criteria, survival times and the presence of mutations. In summary, canine digital melanoma differs from molecular genetic data of canine oral melanoma and human melanoma, especially regarding the proportion of mutations.
与犬口腔黑色素瘤相比，犬趾部黑色素瘤在分子遗传学水平上仍未得到充分研究。本研究的目的是检测趾部黑色素瘤中的突变基因。通过数字液滴PCR（ddPCR）检测了86例犬趾部黑色素瘤石蜡包埋样本中的V595E变体，并检测了……外显子11的突变。此外，通过桑格测序分析了……的外显子2和3。分析了9只犬基因组DNA中……的拷贝数变异（CNV）。未发现V595E变体，在……中，16/70例肿瘤（23%）中发现了一个单核苷酸多态性。……的拷贝数在4至6之间变化。在所检测的86例（25.6%）黑色素瘤中，……外显子2的第12和13密码子发生了突变。在热点位点内还发现了其他相互排斥的……突变，即……外显子3第61密码子：2/55（3.6%）；……外显子2第12和13密码子：2/83（2.4%）；以及……外显子3第61密码子：9/86（10.5%）。然而，组织学恶性标准、生存时间与……突变的存在之间未发现相关性。总之，犬趾部黑色素瘤与犬口腔黑色素瘤和人类黑色素瘤的分子遗传学数据不同，尤其是在……突变的比例方面。
