Department of Hepatobiliary Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi Province, China.
Toxicol In Vitro. 2010 Mar;24(2):538-45. doi: 10.1016/j.tiv.2009.11.008. Epub 2009 Nov 12.
The side population (SP) is commonly applied to analyzing cancer stem cells (CSCs) biology. However, the Hoechst33342 (Hoechst) dye, which is used for SP isolation, is demonstrated to be toxic to cells. The aim of this study was to identify a dye that might replace or offer an alternative to Hoechst use. In the trial group, by FACS binding Rhodamine123 (Rho), Rho(low) (low Rho fluorescence) and Rho(high) (high Rho fluorescence) were sorted from the MHCC97 cell line. In the control group, SP (low Hoechst fluorescence) and NSP (high Hoechst fluorescence) were obtained through Hoechst/FACS. The percentages of subpopulations were as following: Rho(low) 2.1%, Rho(high) 97.9%, SP 1.4%, NSP 98.6%. The proliferative abilities in vitro (growth rate, soft agar clone formation rate), expressions of stem cell markers (early hepatic marker AFP and CSCs marker CD133), and tumorigenicities in vivo were significantly higher in the Rho(low) and SP cells than those in the Rho(high) and NSP cells, respectively (P<0.05). In addition, each result was the same between the Rho(low) and SP cells (P>0.05). Taken together, Rho/FACS may not be as effective as Hoechst/FACS for CSCs sorting, but given its low toxicity and cost, it may be a useful method for CSCs identification.
侧群（SP）常用于分析癌症干细胞（CSC）的生物学特性。然而，用于 SP 分离的 Hoechst33342（Hoechst）染料已被证明对细胞具有毒性。本研究旨在寻找一种可能替代 Hoechst 的染料。在实验组中，通过 FACS 结合 Rhodamine123（Rho），从 MHCC97 细胞系中分离出 Rho(low)（低 Rho 荧光）和 Rho(high)（高 Rho 荧光）。在对照组中，通过 Hoechst/FACS 获得 SP（低 Hoechst 荧光）和 NSP（高 Hoechst 荧光）。亚群的比例如下：Rho(low) 2.1%，Rho(high) 97.9%，SP 1.4%，NSP 98.6%。体外增殖能力（生长速度、软琼脂克隆形成率）、干细胞标志物（早期肝标志物 AFP 和 CSCs 标志物 CD133）的表达以及体内致瘤性在 Rho(low)和 SP 细胞中明显高于 Rho(high)和 NSP 细胞，分别为（P<0.05）。此外，Rho(low)和 SP 细胞之间的每个结果均相同（P>0.05）。综上所述，Rho/FACS 可能不如 Hoechst/FACS 有效用于 CSC 分选，但鉴于其低毒性和低成本，它可能是一种用于 CSC 鉴定的有用方法。
