Transcriptomic profile of canine mammary ductal carcinoma. Transcriptomic profile of canine mammary ductal carcinoma

Mammary neoplasms are the most frequent in non-neutered bitches. Here we used next generation sequencing (NGS), to determine the transcriptional profile of samples of canine mammary ductal carcinomas and to contribute to the clarification of the importance of deregulated molecules in the molecular pathways involved in the disease. For this, samples of tumor tissue from mammary ductal carcinoma (without metastases) and non-tumor breast tissue from radical mastectomy of six bitches of different breeds and ages were used. Overall design: Samples of tumor tissue from mammary ductal carcinoma and adjacent non-tumor mammary tissue, from the radical mastectomy of 6 bitches of different breeds, not neutered, aged between 5 and 14 years, attended by the Small Animal Surgery Service of the Veterinary Hospital.RNA was extracted and was pooled with three samples each, forming four groups (two tumoral and two control samples). Each pool was used to build the four sequencing libraries. The cDNA libraries were built with the TruSeqTM Stranded Total RNA Sample Preparation Kit v2 (Illumina) and sequencing took place on the NextSeq 500 System platform (Illumina, USA) using the NextSeq 500/550 High Output v2 150 cycles (Illumina) Sequencing Kit, according to the manufacturer's instructions.

乳腺肿瘤（Mammary neoplasms）是未绝育母犬最常见的肿瘤性疾病。本研究采用下一代测序技术（Next Generation Sequencing, NGS）解析犬乳腺导管癌样本的转录组表达谱，旨在阐明失调分子在该疾病相关分子通路中的重要性，为相关研究提供支撑。
为此，本研究采集了6只不同品种、不同年龄未绝育母犬的根治性乳房切除术样本，包括无转移的乳腺导管癌肿瘤组织及癌旁正常乳腺组织。
实验整体设计：选取6只年龄5~14岁、未绝育的不同品种母犬，采集其乳腺导管癌肿瘤组织及对应癌旁正常乳腺组织，所有样本均由该兽医院小动物外科服务中心通过根治性乳房切除术获取。
提取样本总RNA后，每3个样本合并为1个混合池，最终构建4组混合样本（2组肿瘤样本池与2组正常对照样本池），每组混合池用于构建1个测序文库，共得到4个测序文库。
按照试剂盒说明书，使用TruSeq™ 链特异性总RNA样本制备试剂盒v2（Illumina）构建cDNA文库；随后依托NextSeq 500测序平台（Illumina，美国），搭配NextSeq 500/550 High Output v2 150循环测序试剂盒（Illumina）完成测序。