Department of Clinical Pharmaceutics, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences and Okayama University Hospital, Okayama.
Department of Hematology, Oncology and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences and Okayama University Hospital, Okayama.
Jpn J Clin Oncol. 2014 Oct;44(10):963-8. doi: 10.1093/jjco/hyu110. Epub 2014 Aug 28.
The echinoderm microtubule associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene was identified in patients with non-small cell lung cancer. To the best of our knowledge, there are only three cell lines harboring the EML4-ALK fusion gene, which have contributed to the development of therapeutic strategies. Therefore, we tried to establish a new lung cancer cell line harboring EML4-ALK.
A 61-year-old Japanese female presented with chest discomfort. She was diagnosed with left lung adenocarcinoma with T4N3M1 Stage IV. Although she was treated with chemotherapy, her disease progressed with massive pleural effusion. Because the EML4-ALK rearrangement was found in a biopsied specimen using fluorescence in situ hybridization, she was treated with crizotinib. She did well for 3 months.
Tumor cells were obtained from the malignant pleural effusion before treatment with crizotinib. Cells continued to proliferate substantially for several weeks. The cell line was designated ABC-11. The EML4-ALK fusion protein and genes were identified in ABC-11 cells using fluorescence in situ hybridization and immunohistochemistry, respectively. ABC-11 cells were sensitive to crizotinib and next-generation ALK inhibitors (ceritinib and AP26113), as determined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Phosphorylated ALK protein and its downstream signaling were suppressed by treatment with crizotinib in western blotting. Furthermore, we could transplant ABC-11 cells subcutaneously into BALB/c nu/nu mice.
We successfully established a new lung adenocarcinoma cell line harboring the EML4-ALK fusion gene. This cell line could contribute to future research of EML4-ALK-positive lung cancer both in vivo and in vitro.
棘皮动物微管相关蛋白样 4（EML4）-间变性淋巴瘤激酶（ALK）融合基因在非小细胞肺癌患者中被发现。据我们所知，仅有三种携带有 EML4-ALK 融合基因的细胞系，这为治疗策略的发展做出了贡献。因此，我们试图建立一种新的携带有 EML4-ALK 的肺癌细胞系。
一名 61 岁的日本女性因胸痛就诊。她被诊断为左侧肺腺癌伴 T4N3M1 Ⅳ期。尽管接受了化疗，但她的疾病仍进展为大量胸腔积液。由于在活检标本中使用荧光原位杂交发现了 EML4-ALK 重排，她接受了克唑替尼治疗。她在 3 个月内病情稳定。
在接受克唑替尼治疗前，从恶性胸腔积液中获得了肿瘤细胞。这些细胞继续大量增殖数周。该细胞系被命名为 ABC-11。使用荧光原位杂交和免疫组织化学分别在 ABC-11 细胞中鉴定出 EML4-ALK 融合蛋白和基因。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定法，ABC-11 细胞对克唑替尼和下一代 ALK 抑制剂（塞瑞替尼和 AP26113）敏感。在 Western blot 中，用克唑替尼处理可抑制磷酸化 ALK 蛋白及其下游信号转导。此外，我们可以将 ABC-11 细胞皮下移植到 BALB/c nu/nu 小鼠中。
我们成功建立了一种新的携带有 EML4-ALK 融合基因的肺腺癌细胞系。该细胞系可有助于未来在体内和体外对 EML4-ALK 阳性肺癌的研究。
