肝癌细胞系Hep3B中CD133+细胞亚群的分离及其特性的研究

目的研究CD133在人肝癌细胞系Hep3B中的表达以及CD133+细胞的体外增殖、自我更新及体内成瘤能力，初步探讨肝癌中CD133+细胞亚群的干细胞特性。方法流式细胞仪检测未分选的Hep3B细胞中CD133+细胞表达情况；免疫磁珠分选技术纯化CD133+肿瘤细胞；MTT法检测CD133+细胞体外增殖能力；无血清培养纯化...

Isolation of CD133 + Cell and Stem Cell Property from Hepatocarcinoma Hep3B Cell Line

Objective To detect the expression of CD133 in hepatocellular carcinoma Hep3 B cell line, detect proliferative and self-renew ability of CD133 ＋ tumor cells in vitro, implantation experiment of CD133 ＋ 3 B cell in vivio, and study its stem-cell property. Methods Flow eytometry was used to detect the expression of putative tumor-initiated cell marker CD133 in Hep3B cell line. The isolation technique with immunomagnetie beads was applied to purify CD133 ＋ cells. The proliferative ability of CD133 ＋ tumor cells was estimated by MTY assay in vitro. The proliferative ability of CD133 ＋ , CD133 - and control Hep3B cells were statistically compared. CD133 ＋ tumor cells were cultured in serum-free medium after isolation and observed. Tumor cells implantation experiments were done to test the tumorigenicity of CD133 ＊ , CD133 - and control Hep3B cells. Results 9.13% CD133 ＋ cells were detected in Hep3B cell line. In serum-free RPMI 1640, their UV opti- cal of the CD133 ＋ tumor cells was 0. 311,0. 504,0. 602 and 0. 800 at 1,3,5 and 7 days respectively. Compared with CD133 - cells and control Hep3B cells, CD133 ＋ cells demonstrated increased proliferation capacity. Tumor cell implantation experiments showed that only 1×10^3 CD133 ＋ cells were sufficient for tumor formation,whereas an injection of 1×10^5 CD133 - cells could initiate tumors. Conclusion CD133 ＋ cells subpopulation has stronger ability to proliferate in vitro and to form tumors in vivo than other subpopulations in Hep3B cell line. In general, CD133 ＋ cell subpopulation has stem-cell property.