The in-vitro spheroid culture induces a more highly differentiated but tumorigenic population from melanoma cell lines
The in-vitro spheroid culture induces a more highly differentiated but tumorigenic population from melanoma cell lines
复制标题
体外球状培养物从黑色素瘤细胞系中诱导出高度分化但致瘤的细胞群
DOI：
10.1097/cmr.0b013e32836314e3
复制
发表时间：
2013-08-01
影响因子：
2.2
通讯作者：
Sun, Ran
中科院分区：
文献类型：
--
作者：
Mo, Jing;Sun, Baocun;Sun, Ran
癌症干细胞 (CSC) 已在各种恶性肿瘤中被发现，并且已检查了不同的特性来表征 CSC：免疫功能低下小鼠的致瘤性、干细胞表面标记、标记保留特性以及作为非粘附球体的增殖。本研究探讨了这些方法之间的一致性和效率。在检查的黑色素瘤细胞系（A375、A875、MUM-2b 和 MUM-2c）中，只有 A375 和 MUM-2c 以非贴壁球体形式生长，并在体外在确定的无血清培养基中持续增殖。流式细胞术和免疫荧光分析表明，与贴壁细胞相比，球体来源的细胞中表达黑色素瘤干细胞候选表面标记物（如ABCB5、CD133、CD20和CD271）的细胞比例较小，而表达黑素细胞分化标记物（如HMB45和S100蛋白）的细胞比例较大。令人惊讶的是，与亲代细胞相比，高度分化的球体来源的黑色素瘤细胞在体内表现出更高的致瘤潜力，这通过更短的肿瘤孵育时间（A375）和更少数量的启动肿瘤形成所需的细胞（A375和MUM-2c）来表明。尽管组织病理学特征相似，但表达谱表明，相对于干或通过免疫组织化学分析检测到的分化标记物，源自球源性黑色素瘤细胞的异种移植物表现出更具致瘤性的表型。因此，非贴壁培养物中的球体形成可能不是根据候选标记富集黑色素瘤干细胞的首选替代体外方法，但可能是激活潜在 CSC 的有利条件。黑色素瘤研究 23:254-263 (C) 2013 Wolters Kluwer Health 垂直条 Lippincott Williams & Wilkins。
Cancer stem cells (CSCs) have been identified in various malignancies, and different properties have been examined to characterize CSCs: tumorigenicity in immunocompromised mice, stem cell surface markers, label-retaining properties, and proliferation as nonadherent spheres. This study explored the consistency and efficiency among these methods. Among the melanoma cell lines examined (A375, A875, MUM-2b, and MUM-2c), only A375 and MUM-2c grew as nonadherent spheres and continuously propagated in a defined serum-free medium in vitro. Flow cytometry and immunofluorescence analysis indicated that sphere-derived cells contained a smaller proportion of cells expressing the candidate surface markers of melanoma stem cells such as ABCB5, CD133, CD20 and CD271, and a larger proportion of cells expressing melanocytic differentiation markers such as HMB45 and S100 protein, compared with adherent cells. Surprisingly, the more highly differentiated sphere-derived melanoma cells exhibited increased tumorigenic potential in vivo, as indicated by shorter tumor incubation (A375) and smaller number of cells required to initiate tumor formation (A375 and MUM-2c) compared with those of parental cells. Despite the similarity in histopathological characteristics, the expression profile indicated that xenografts derived from sphere-derived melanoma cells exhibited a more tumorigenic phenotype with respect to the stem or the differentiation markers detected by immunohistochemical analysis. Therefore, sphere formation in nonadherent cultures may not be a preferred surrogate in-vitro method for enriching melanoma stem cells according to candidate markers but may be a favorable condition for activating potential CSCs. Melanoma Res 23:254-263 (C) 2013 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.