Department of Ophthalmology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, No. 12, Lane 833, Zhizaoju Road, Huangpu District, Shanghai, 200001, China; Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, No. 12, Lane 833, Zhizaoju Road, Huangpu District, Shanghai, 200001, China.
Department of Ophthalmology, Leiden University Medical Center, PO BOX 9600, 2300RC, Leiden, The Netherlands; Department of Ophthalmology, The Second Hospital of Jilin University, No. 218, Ziqiang Road, Nanguan District, Changchun, China.
Exp Eye Res. 2018 Aug;173:51-63. doi: 10.1016/j.exer.2018.03.030. Epub 2018 Apr 10.
Conjunctival melanoma (CM) is associated with metastases formation, can be fatal, and occurs in all different races. While cell lines are essential for experimental research, all available CM cell lines are derived from Caucasian patients. Furthermore, they are not derived from metastases. We aimed to establish a new CM cell line from a parotid metastasis in a Han Chinese patient and to depict its characteristics. The novel cell line, CM-AS16, was obtained from a surgical parotid sample and determined as a unique one with short tandem repeat (STR) analysis. It has been successively sub-cultured in vitro for more than 100 passages and exhibits rapid proliferation and migration. Chromosome analysis shows abundant chromosome aberrations, while whole exome sequencing (WES) reveals a typical NRAS mutation (Q61R). In vivo tumor growth was successfully established in a NOD/SCID mice model, and the immunophenotypes, such as HMB45, Melan A, S100, SOX10 and Ki67, manifested similar between the original tumor and the xenograft by immunohistochemistry. A MEK inhibitor binimetinib prominently suppressed in vitro cell growth by inhibiting ERK1/2 phosphorylation. In addition, monoclonal cells were used to demonstrate the drug sensitivity of different cells. In conclusion, the first cell line, CM-AS16, that is derived from a CM in a Han Chinese patient has highly malignant characteristics and a typical NRAS mutation. It may be used as a tool for further exploration of the molecular mechanisms of CM.
结膜黑色素瘤（CM）与转移形成有关，可能致命，发生于所有不同种族。虽然细胞系对于实验研究至关重要，但所有现有的 CM 细胞系均源自白种人患者，而且并非源自转移灶。我们旨在从一名汉族患者的腮腺转移灶中建立新的 CM 细胞系，并描绘其特征。新型细胞系 CM-AS16 源自手术切除的腮腺样本，通过短串联重复序列（STR）分析确定其为独特的细胞系。它已在体外成功连续传代培养超过 100 代，表现出快速增殖和迁移的特性。染色体分析显示大量染色体畸变，而全外显子组测序（WES）揭示了典型的 NRAS 突变（Q61R）。在 NOD/SCID 小鼠模型中成功建立体内肿瘤生长模型，免疫组织化学染色显示，原始肿瘤和异种移植物之间的免疫表型（如 HMB45、Melan A、S100、SOX10 和 Ki67）相似。MEK 抑制剂 binimetinib 通过抑制 ERK1/2 磷酸化显著抑制体外细胞生长。此外，使用单克隆细胞证明了不同细胞对药物的敏感性。总之，源自汉族 CM 患者的首个 CM-AS16 细胞系具有高度恶性特征和典型的 NRAS 突变。它可能被用作进一步探索 CM 分子机制的工具。
