Identification of side population cells from bladder cancer cells by dyecycle violet staining
Identification of side population cells from bladder cancer cells by dyecycle violet staining
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通过染料环紫染色鉴定膀胱癌细胞的侧群细胞
DOI：
10.4161/cbt.7.10.6637
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发表时间：
2008-10-01
影响因子：
3.6
通讯作者：
Che, Xiang Ming
中科院分区：
文献类型：
--
作者：
She, Jun Jun;Zhang, Peng Ge;Che, Xiang Ming
侧群 (SP) 细胞可用于识别假定的癌症干细胞 (CSC)，但该技术因需要紫外 (UV) 激光源而受到阻碍。 DyeCycle Violet 试剂 (DCV) 是一种 DNA 结合染料，可用于配备普通紫色激光二极管 (VLD) 的流式细胞仪。在本文中，我们使用 Hoechst 33342 或 DCV 染色分析了几种膀胱癌细胞系的 SP 细胞。 Hoechst 33342 染色的 SP 细胞使用配备 UV 的流式细胞仪进行鉴定，而 DCV 染色的 SP 细胞则使用配备 VLD 的流式细胞仪进行鉴定。 DCV 染色还用于从 SW780 细胞中分选 SP 和非 SP (NSP) 细胞。进一步分析表明SP细胞可以产生SP和NSP细胞。 SP细胞的集落形成能力明显强于NSP细胞。当注射到裸鼠体内时，只要 1 x 103 个 SP 细胞就可以在 12 个注射部位中的 8 个部位引发肿瘤。相比之下，将NSP细胞注射到裸鼠体内未能引发肿瘤。 RT-PCR数据显示SP和NSP细胞之间ABCG2、MDRI、Bmi-1和Oct-4的表达存在差异，表明SP细胞具有一些干细胞特征。我们得出结论，通过 DCV 染色鉴定的 SP 细胞能够不对称分裂、自我更新和肿瘤起始。我们的研究还表明 DCV 是鉴定 SP 细胞的有用试剂。
Side population (SP) cells can be used to identify putative cancer stem cells (CSC), but this technique is hampered by the requirement for an ultraviolet (UV) laser source. DyeCycle Violet reagent (DCV) is a DNA-binding dye that can be used in the common violet laser diode (VLD)-equipped flow cytometer. In this paper, we analyzed SP cells from several bladder cancer cell lines using either Hoechst 33342 or DCV staining. The Hoechst 33342-stained SP cells were identified with a UV-equipped flow cytometer, while the DCV-stained SP cells were identified with a VLD-equipped flow cytometer. DCV staining was also used to sort SP and non-SP (NSP) cells from SW780 cells. Further analysis revealed that SP cells could give rise to both SP and NSP cells. The colony-forming ability of SP cells was significant greater than that of NSP cells. When injected into nude mice, as few as 1 x 103 SP cells could initiate tumors in eight of twelve injection sites. In contrast, the injection of NSP cells into nude mice failed to initiate tumors. RT-PCR data showed that the expression of ABCG2, MDRI, Bmi-1 and Oct-4 differed between SP and NSP cells, suggesting that SP cells possess some stem cell characteristics. We conclude that SP cells identified by DCV staining are capable of asymmetric division, self-renewal and tumor initiation. Our study also indicates that DCV is a useful reagent for the identification of SP cells.