Establishment of a tumor sphere cell line from a metastatic brain neuroendocrine tumor.
Establishment of a tumor sphere cell line from a metastatic brain neuroendocrine tumor.
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从转移性脑神经内分泌肿瘤建立肿瘤球细胞系。
DOI：
10.1007/s00795-017-0160-0
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发表时间：
2017
期刊：
影响因子：
1.8
通讯作者：
Asai A.
中科院分区：
文献类型：
--
作者：
Iwata R;Maruyama M;Ito T;Nakano Y;Kanemura Y;Koike T;Oe S;Yoshimura K;Nonaka M;Nomura S;Sugimoto T;Yamada H;Asai A.
神经内分泌肿瘤是一种罕见的肿瘤，人们对这种疾病中肿瘤干细胞的存在知之甚少。确定致瘤人群将有助于开发针对神经内分泌肿瘤的有效治疗方法。手术切除的原发不明来源神经内分泌肿瘤脑转移灶分离并在无血清神经球培养基中培养。研究了干细胞的特性，包括自我更新、分化潜力和干细胞标志物表达。采用颅内异种移植模型评估肿瘤形成情况。用细胞活力法测定替莫唑胺的体外作用。我们建立了神经内分泌肿瘤球细胞系anti - 27s，该细胞系表现出稳定的指数生长，在体外几乎无限制地扩增，并表达CD133、nestin、Sox2和醛脱氢酶等干细胞标志物。fbs诱导的分化降低Sox2和nestin的表达。基于实时荧光定量PCR， ANI-27S细胞表达神经内分泌标志物synaptophysin和chromogranin A.颅内异种移植脑瘤再现了患者原发肿瘤，替莫唑胺对肿瘤球细胞表现出细胞毒性作用。我们首次证实了原发性神经内分泌肿瘤脑转移灶中存在球状干细胞样细胞群。我们还证明了替莫唑胺对这种疾病的潜在治疗效果。
Neuroendocrine tumors are rare, and little is known about the existence of cancer stem cells in this disease. Identification of the tumorigenic population will contribute to the development of effective therapies targeting neuroendocrine tumors. Surgically resected brain metastases from a primary neuroendocrine tumor of unknown origin were dissociated and cultured in serum-free neurosphere medium. Stem cell properties, including self-renewal, differentiation potential, and stem cell marker expression, were examined. Tumor formation was evaluated using intracranial xenograft models. The effect of temozolomide was measured in vitro by cell viability assays. We established the neuroendocrine tumor sphere cell line ANI-27S, which displayed stable exponential growth, virtually unlimited expansion in vitro, and expression of stem-cell markers such as CD133, nestin, Sox2, and aldehyde dehydrogenase. FBS-induced differentiation decreased Sox2 and nestin expression. On the basis of real-time PCR, ANI-27S cells expressed the neuroendocrine markers synaptophysin and chromogranin A. Intracranial xenotransplanted brain tumors recapitulated the original patient tumor and temozolomide exhibited cytotoxic effects on tumor sphere cells. For the first time, we demonstrated the presence of a sphere-forming, stem cell-like population in brain metastases from a primary neuroendocrine tumor. We also demonstrated the potential therapeutic effects of temozolomide for this disease.