Isolation and characterization of cancer stem-like side population cells in human oral cancer cells
Isolation and characterization of cancer stem-like side population cells in human oral cancer cells
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DOI：
10.1016/j.oraloncology.2011.06.501
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发表时间：
2011-09-01
期刊：
影响因子：
4.8
通讯作者：
Umeda, Masahiro
中科院分区：
文献类型：
--
作者：
Yanamoto, Souichi;Kawasaki, Goro;Umeda, Masahiro
最近的研究表明，癌症干细胞可能是肿瘤发生的原因，并有助于某些个体对癌症治疗的抵抗。一些研究表明，从不同的癌细胞系分离的侧群（SP）细胞具有干细胞样的特性，然而，很少有研究SP细胞在人类口腔癌中的作用的报告。为了确定人口腔癌细胞系是否含有SP细胞部分，我们首先通过荧光激活细胞分选分离SP细胞，然后使用SCC 25舌癌细胞系在无血清培养基（SFM）中培养，使得SP细胞能够增殖以保持CSC性质。通过RT-PCR检测SP细胞和非SP细胞中干细胞标志物（ABCG 2、Oct-4和EpCAM）的差异表达谱。MTT法检测5-FU对细胞生长的抑制作用。克隆形成试验评价克隆形成能力。SCC-25细胞含0.23%的SP细胞。SP细胞的部分可用于在SFM培养物中生长。与非SP细胞相比，SP细胞显示出更高的干细胞标志物（ABCG 2、Oct-4和EpCAM）的mRNA表达。此外，与非SP细胞相比，SP细胞对5-FU表现出更高的耐药性。在相同细胞数下，SP细胞的克隆形成率显著高于非SP细胞（P < 0.01）。我们从口腔癌细胞系中分离出癌干细胞样SP细胞。SP细胞具有肿瘤干细胞的特性、耐药性和高增殖能力。进一步表征癌症干细胞样SP细胞可能为新的治疗靶点提供新的见解。(C)2011爱思唯尔有限公司保留所有权利。
Recent studies suggest that cancer stem cells may be responsible for tumorigenesis and contribute to some individuals' resistance to cancer therapy. Some studies demonstrate that side population (SP) cells isolated from diverse cancer cell lines harbor stem cell-like properties; however, there are few reports examining the role of SP cells in human oral cancer. To determine whether human oral cancer cell lines contain a SP cell fraction, we first isolated SP cells by fluorescence activated cell sorting, followed by culturing in serum-free medium (SFM) using the SCC25 tongue cancer cell line, so that SP cells were able to be propagated to maintain the CSC property. Differential expression profile of stem cell markers (ABCG2, Oct-4 and EpCAM) was examined by RT-PCR in either SP cells or non-SP cells. Growth inhibition by 5-FU was determined by the MTT assay. Clonogenic ability was evaluated by colony formation assay. SCC25 cells contained 0.23% SP cells. The fraction of SP cells was available to grow in SFM cultures. SP cells showed higher mRNA expression of stem cell markers (ABCG2, Oct-4 and EpCAM) as compared with non-SP cells. Moreover, SP cells demonstrated more drug resistance to 5-FU, as compared with non-SP cells. The clone formation efficiency of SP cells was significantly higher than non-SP cells at an equal cell number (P < 0.01). We isolated cancer stem-like SP cells from an oral cancer cell line. SP cells possessed the characteristics of cancer stem cells, chemoresistance, and high proliferation ability. Further characterization of cancer stem-like SP cells may provide new insights for novel therapeutic targets. (C) 2011 Elsevier Ltd. All rights reserved.