Department of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI 48109, USA.
Dept of Medicine, University of Chicago, USA.
Oral Oncol. 2018 Dec;87:21-28. doi: 10.1016/j.oraloncology.2018.10.012. Epub 2018 Oct 18.
Limited availability of validated human adenoid cystic carcinoma (ACC) cell lines has hindered the mechanistic understanding of the pathobiology of this malignancy and the development of effective therapies. The purpose of this work was to generate and characterize a human ACC cell line.
Immediately after surgery, a tumor fragment from a minor salivary gland from the tongue of a female Caucasian was minced, dissociated, and a single cell suspension was plated in fibronectin-coated flasks. A culture medium containing bovine brain extract and rhEGF was optimized for these cells. Whole exome sequencing was used to evaluate the presence of MYB-NFIB translocation.
The University of Michigan-Human Adenoid Cystic Carcinoma (UM-HACC)-2A cells showed continuous growth in monolayers for at least 180 in vitro passages while maintaining epithelial morphology. Short-tandem repeat (STR) profiling confirmed a 100% match to patient DNA. Whole exome sequencing revealed the presence of the MYB-NFIB fusion in UM-HACC-2A cells, which was confirmed by PCR analysis. Western blots revealed high expression of epithelial markers (e.g. E-cadherin, EGFR, pan-cytokeratin) and proteins associated with ACC (e.g. c-Myb, p63). Developmental therapeutic studies showed that UM-HACC-2A cells were resistant to cisplatin (IC = 44.7 µM) while more responsive to paclitaxel (IC = 0.0006 µM). In a pilot study, we observed that UM-HACC-2A cells survived orthotopic transplantation into the submandibular gland. Notably, one of the mice injected with UM-HACC-2A cells exhibited lung metastasis after 6 months.
UM-HACC-2A is a MYB-NFIB fusion-positive ACC cell line that is suitable for mechanistic and developmental therapeutics studies.
可用的经过验证的人腺样囊性癌 (ACC) 细胞系有限，这阻碍了对这种恶性肿瘤的病理生物学的机制理解，以及有效治疗方法的发展。本工作的目的是生成并鉴定人腺样囊性癌细胞系。
手术切除后，立即将来自舌部小唾液腺的肿瘤组织切成小块，分离，制成单细胞悬液，接种于纤维连接蛋白包被的培养瓶中。优化了含有牛脑提取物和 rhEGF 的培养基，用于这些细胞。全外显子组测序用于评估 MYB-NFIB 易位的存在。
密歇根大学人腺样囊性癌（UM-HACC）-2A 细胞在至少 180 代体外培养中保持单层连续生长，同时保持上皮形态。短串联重复（STR）分析证实 100%与患者 DNA 匹配。全外显子组测序显示 UM-HACC-2A 细胞中存在 MYB-NFIB 融合，通过 PCR 分析得到证实。Western blot 显示上皮标志物（如 E-钙黏蛋白、EGFR、泛细胞角蛋白）和与 ACC 相关的蛋白（如 c-Myb、p63）高表达。发展治疗研究表明，UM-HACC-2A 细胞对顺铂（IC=44.7µM）耐药，而对紫杉醇（IC=0.0006µM）更敏感。在一项初步研究中，我们观察到 UM-HACC-2A 细胞在颌下腺原位移植后存活。值得注意的是，其中一只注射 UM-HACC-2A 细胞的小鼠在 6 个月后出现肺转移。
UM-HACC-2A 是人腺样囊性癌，具有 MYB-NFIB 融合阳性，适合机制和发展治疗学研究。
