Identification of Cancer Stem-like Side Population Cells in Ovarian Cancer Cell Line OVCAR-3
Identification of Cancer Stem-like Side Population Cells in Ovarian Cancer Cell Line OVCAR-3
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DOI：
10.1080/01913120903086072
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发表时间：
2009-01-01
影响因子：
1
通讯作者：
Suo, Zhenhe
中科院分区：
文献类型：
--
作者：
Gao, Quanli;Geng, Li;Suo, Zhenhe
侧群（SP）细胞可以在许多正常和恶性组织中富集干细胞样细胞。然而，SP方法的应用引起了干细胞研究领域的特别关注，并且SP细胞在细胞培养中的存在正在争论，最可能的是因为不同的细胞系需要不同的技术修改，特别是当考虑细胞染色时。在这项研究中，作者的目的是揭示hoechst 33342染色是否需要广泛的优化，以识别人卵巢癌细胞系OVCAR-3中的SP细胞。经系统评价，发现2.5g/mLHoechst 33342染色60 min即可获得理想的SP群体，其占整个细胞群体的0.9%。分选后的SP细胞集落形成效率显著高于非侧群（non-side population，NSP）细胞，且只有SP细胞能形成全克隆。Real-time PCR结果显示SP细胞中Oct 3/4基因的表达水平高于NSP细胞，提示SP细胞中可能存在肿瘤干细胞。结果强调了细胞研究中SP方法优化的必要性，并且当考虑癌症干细胞鉴定和分离时，该细胞系中的SP细胞值得进一步研究。
Side population (SP) cells may enrich stem-like cells in many normal and malignant tissues. However, SP method application has drawn special attention to the field of stem cell research, and the existence of SP cells in cell culture is being debated, most probably because different cell lines require different technical modifications, especially when cell staining is considered. In this study, the authors aimed to disclose whether the hoechst33342 staining required extensive optimization for identifying SP cells in the human ovarian cancer cell line OVCAR-3. After systematic evaluations, it was found that only 2.5 g/mL hoechst33342 staining of the cells for 60 min could get an ideal SP population, which accounted for 0.9% of the whole cell population. The sorted SP cells showed significantly higher colony formation efficiency than the non-side population (NSP) cells, and only the SP cells could form holoclones. Real-time PCR disclosed that SP cells expressed higher levels of ostemnesso gene Oct3/4 than the NSP cells did, indicating that the SP cells might harbor cancer stem cells in this cell line. The results highlight the necessity of SP method optimization in cell studies, and the SP cells in this cell line merit further studies when cancer stem cell identification and isolation are considered.