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犬乳腺肿瘤细胞系的建立及部分特性研究

Establishment and Partial Characterization of Canine Mammary Tumor Cell Lines.

作者信息

Vazquez Eliza, Dominguez Luis, Silverio Brian, Torres Geobanni, Garibay-Escobar Adriana, Queiroga Felisbina Luisa, Velazquez Carlos

机构信息

Department of Chemistry-Biology, University of Sonora, Blvd. Luis Encinas y Rosales s/n, Hermosillo 83000, Mexico.

Veterinary and Animal Research Centre (CECAV), University of Trás-os-Montes and Alto Douro, 5001-801 Vila Real, Portugal.

出版信息

Animals (Basel). 2025 Jul 7;15(13):1991. doi: 10.3390/ani15131991.

Abstract

Mammary tumors are the most common neoplasms diagnosed in female dogs and have been considered excellent models for studying human breast cancer. Establishing cell lines from primary cultures of canine mammary tumors provides an in vitro model to better understand the disease and develop new treatments. This study aimed to establish and characterize canine mammary tumor cell lines. Ten cell cultures were generated from tumor tissue obtained from affected dogs, including seven from primary mammary tumors and three from metastatic sites. Characterization included molecular marker expression (ER, PR, HER2, cytokeratin 5/6 (CK5/6), vimentin, and the marker of cell proliferation Ki67) and in vitro tumorigenic capacity assessment. Additionally, the susceptibility of five cell lines to DOX, 5-FU, paclitaxel, colchicine, and carboplatin was evaluated using the MTT assay. ICC analysis revealed negative expression of hormonal receptors (ER and PR) in five cell lines, while only one cell line was positive for both. Six cell lines were HER2-negative and positive for vimentin. Five cell lines exhibited in vitro tumorigenic capacity, forming colonies in soft agar. DOX showed the highest growth-inhibitory effect (DOX > Paclitaxel > Colchicine > 5-FU > Carboplatin). Two cell lines had a minimal concentration for 50% inhibition in vitro (IC) < 0.63 µM and 4.37 ± 0.40 µM for DOX, while one was sensitive to colchicine and paclitaxel (IC 0.19 µM and 0.04 µM, respectively). All tested cell lines were resistant to carboplatin and 5-FU. These cell lines provide a valuable model for studying breast cancer in humans and dogs and evaluating new potential therapeutic strategies.

摘要

乳腺肿瘤是雌性犬类中最常见的被诊断出的肿瘤,并且一直被认为是研究人类乳腺癌的优秀模型。从犬乳腺肿瘤原代培养物中建立细胞系提供了一个体外模型,以更好地理解该疾病并开发新的治疗方法。本研究旨在建立并表征犬乳腺肿瘤细胞系。从患病犬获取的肿瘤组织中生成了10个细胞培养物,其中7个来自原发性乳腺肿瘤,3个来自转移部位。表征包括分子标志物表达(雌激素受体(ER)、孕激素受体(PR)、人表皮生长因子受体2(HER2)、细胞角蛋白5/6(CK5/6)、波形蛋白以及细胞增殖标志物Ki67)和体外致瘤能力评估。此外,使用MTT法评估了5个细胞系对阿霉素(DOX)、5-氟尿嘧啶(5-FU)、紫杉醇、秋水仙碱和卡铂的敏感性。免疫细胞化学(ICC)分析显示,5个细胞系中的激素受体(ER和PR)呈阴性表达,而只有1个细胞系两者均为阳性。6个细胞系HER2呈阴性,波形蛋白呈阳性。5个细胞系表现出体外致瘤能力,在软琼脂中形成集落。DOX显示出最高的生长抑制作用(DOX>紫杉醇>秋水仙碱>5-FU>卡铂)。2个细胞系的体外50%抑制最低浓度(IC)<0.63 μM,DOX为4.37±0.40 μM,而1个细胞系对秋水仙碱和紫杉醇敏感(IC分别为0.19 μM和0.04 μM)。所有测试的细胞系对卡铂和5-FU均耐药。这些细胞系为研究人类和犬类乳腺癌以及评估新的潜在治疗策略提供了有价值的模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a22/12249022/a151e75675ca/animals-15-01991-g001.jpg

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