Cancer stem-like cells can be isolated with drug selection in human ovarian cancer cell line SKOV3
Cancer stem-like cells can be isolated with drug selection in human ovarian cancer cell line SKOV3
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DOI：
10.1093/abbs/gmq067
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发表时间：
2010-09-01
影响因子：
3.7
通讯作者：
Guo, Lihe
中科院分区：
文献类型：
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作者：
Ma, Li;Lai, Dongmei;Guo, Lihe
一种新出现的耐药肿瘤发展模型利用了一群自我更新的恶性祖细胞，称为癌症干细胞（CSC）或癌症起始细胞（CIC）。本研究的目的是从卵巢癌细胞系SKOV 3中增殖这样的CIC。在补充有表皮生长因子、碱性成纤维细胞生长因子、白血病抑制因子和胰岛素的无血清培养系统或标准含血清系统中，使用40.0 μ mol/l顺铂和10.0 μ mol/l紫杉醇选择SKOV 3球形细胞。这些细胞在药物选择（顺铂和紫杉醇）和无血清培养系统中形成非粘附球。所选的球形细胞对顺铂、紫杉醇、阿霉素和甲氨蝶呤更耐药。重要的是，球形细胞具有自我更新的特性，高表达干细胞基因Nanog，Oct 4，sox 2，nestin，ABCG 2，CD 133和干细胞因子受体CD 117（c-kit）。流式细胞仪检测结果显示，球形细胞中CD 133（+）/CD 117（+）阳性细胞的比例（71%）明显高于分化细胞（33%）。此外，SKOV 3球细胞更具致瘤性。此外，cDNA微阵列和随后的本体分析显示，大部分的分类基因与血管生成，细胞外基质，整合素介导的信号通路，细胞粘附，细胞增殖。在这种培养系统下从SKOV 3细胞系分离的亚群提供了一个合适的体外模型，用于研究卵巢CSC的存活、自我更新和化学抗性，以及用于开发特异性干扰卵巢CSC的治疗药物。
One emerging model for the development of drug-resistant tumors utilizes a pool of self-renewing malignant progenitors known as cancer stem cells (CSCs) or cancer-initiating cells (CICs). The purpose of this study was to propagate such CICs from the ovarian cancer cell line SKOV3. The SKOV3 sphere cells were selected using 40.0 mu mol/l cisplatin and 10.0 mu mol/l paclitaxel in serum-free culture system supplemented with epidermal growth factor, basic fibroblast growth factor, leukemia inhibitory factor, and insulin or standard serum-containing system. These cells formed non-adherent spheres under drug selection (cisplatin and paclitaxel) and serum-free culture system. The selected sphere cells are more resistant to cisplatin, paclitaxel, adriamycin, and methotrexate. Importantly, the sphere cells have the properties of self-renewal, with high expression of the stem cell genes Nanog, Oct4, sox2, nestin, ABCG2, CD133, and the stem cell factor receptor CD117 (c-kit). Consistently, flow cytometric analysis revealed that the sphere cells have a much higher percentage of CD133(+)/CD117(+)-positive cells (71%) than differentiated cells (33%). Moreover, the SKOV3 sphere cells are more tumorigenic. Furthermore, cDNA microarray and subsequent ontological analyses revealed that a large proportion of the classified genes were related to angiogenesis, extracellular matrix, integrin-mediated signaling pathway, cell adhesion, and cell proliferation. The subpopulation isolation from the SKOV3 cell line under this culture system offers a suitable in vitro model for studying ovarian CSCs in terms of their survival, self-renewal, and chemoresistance, and for developing therapeutic drugs that specifically interfere with ovarian CSCs.