Jurutha Juthathip, Piyasanti Yanika, Sritabtim Kornkanok, Chaipipat Suparat, Siripattarapravat Kannika, Prukudom Sukumal, Jermnak Usuma, Sinsiri Rungthiwa, Wongsuppabut Kakanang, Wongsali Charuwan, Niyatiwatchanchai Nutawan, Sutthiprapa Wijit, Swainson Napachanok, Supsavhad Wachiraphan
Center for Veterinary Diagnostic Laboratory, Bangkhen, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, 10900, Thailand.
Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, 10900, Thailand.
Vet World. 2025 Jun;18(6):1725-1740. doi: 10.14202/vetworld.2025.1725-1740. Epub 2025 Jun 26.
Canine mammary cancer (CMC) is the most frequently diagnosed malignancy in female dogs, sharing significant pathological and molecular similarities with human breast cancer (HBC). Despite the availability of various CMC cell lines, most represent triple-negative orepidermal growth factor receptor 2 (ErbB2)-enriched subtypes, which limit research on hormone receptor-positive cancers. This study aimed to establish and characterize novel CMC cell lines representing luminal A and B subtypes.
Between 2020 and 2021, 31 canine mammary tumors (CMTs) were collected from clinical cases. Tumor tissues were processed for primary culture, and two cell lines - CMGT_071020 and CMGT_180321 - were successfully established. Immunohistochemistry (IHC) was used to assess expression of estrogen receptor alpha (ERα), progesterone receptor (PR), ErbB2, Ki-67, vimentin, and multi-cytokeratin. Functional assays (wound-healing and transwell migration) assessed metastatic behavior. Gene expression (, , , , , , and ) was analyzed using reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Cell line authentication was confirmed through short tandem repeat (STR) profiling and mycoplasma testing.
The CMGT_071020 (luminal B) and CMGT_180321 (luminal A) cell lines were derived from malignant epithelial tumors and maintained stable growth over 30 passages. IHC confirmed molecular subtype classifications. CMGT_071020 exhibited a fibroblast-like morphology, a high Ki-67 index (67%), and superior migratory capacity compared to CMGT_180321 and the commercial ErbB2-enriched REM134 cell line. expression was significantly elevated in CMGT_071020 ( < 0.05), whereas the expression levels of other genes were comparable. STR analysis verified their genetic uniqueness, and both lines were free from mycoplasma contamination.
This study successfully established and characterized two novel hormone receptor-positive CMC cell lines, representing luminal A and luminal B subtypes. The CMGT_071020 line exhibited higher metastatic potential, offering a promising model for aggressive hormone-responsive CMC. These cell lines provide valuable tools for comparative oncology and may facilitate subtype-specific therapeutic research.
犬乳腺肿瘤（CMC）是雌性犬中最常被诊断出的恶性肿瘤，与人类乳腺癌（HBC）在病理和分子方面有显著相似性。尽管有多种CMC细胞系可供使用，但大多数代表三阴性或富含表皮生长因子受体2（ErbB2）的亚型，这限制了对激素受体阳性癌症的研究。本研究旨在建立并表征代表管腔A型和B型亚型的新型CMC细胞系。
在2020年至2021年期间，从临床病例中收集了31例犬乳腺肿瘤（CMT）。对肿瘤组织进行原代培养，成功建立了两个细胞系——CMGT_071020和CMGT_180321。免疫组织化学（IHC）用于评估雌激素受体α（ERα）、孕激素受体（PR）、ErbB2、Ki-67、波形蛋白和多细胞角蛋白的表达。功能试验（伤口愈合和Transwell迁移）评估转移行为。使用逆转录定量实时聚合酶链反应（RT-qPCR）分析基因表达（、、、、、和）。通过短串联重复序列（STR）分析和支原体检测确认细胞系鉴定。
CMGT_071020（管腔B型）和CMGT_180321（管腔A型）细胞系源自恶性上皮肿瘤，并在30代以上保持稳定生长。IHC证实了分子亚型分类。与CMGT_180321和商业化的富含ErbB2的REM134细胞系相比，CMGT_071020表现出成纤维细胞样形态、高Ki-67指数（67%）和更强的迁移能力。在CMGT_071020中表达显著升高（<0.05），而其他基因的表达水平相当。STR分析验证了它们的遗传独特性，且两个细胞系均无支原体污染。
本研究成功建立并表征了两个代表管腔A型和B型亚型的新型激素受体阳性CMC细胞系。CMGT_071020细胞系表现出更高的转移潜能，为侵袭性激素反应性CMC提供了一个有前景的模型。这些细胞系为比较肿瘤学提供了有价值的工具，并可能促进亚型特异性治疗研究。
