人HTB-1膀胱癌细胞系中侧群细胞的生物学功能研究

Biological research on side population cells from human bladder cancer cell line HTB-1

目的从人膀胱癌HTB-1细胞系分选出侧群细胞(SP)并研究其生物学特征。方法人膀胱癌HTB-1细胞培养后分析SP含量,分选出高纯度SP与非侧群细胞(NSP)。分别体外扩增后再分析SP含量。软琼脂克隆集落生成实验比较SP与NSP的体外单克隆生长能力。悬浮肿瘤球实验比较SP与NSP自我更新能力。裸鼠移植实验比较SP与NSP的成瘤性。结果人膀胱癌HTB-1细胞的SP比率为0.93%。SP可产生SP与NSP细胞。SP体外单克隆生长率显著大于NSP(P<0.01)。SP体外悬浮肿瘤球生长率显著大于NSP细胞(P<0.01)。最少种植1 000个SP细胞在裸鼠形成移植性肿瘤。结论 SP细胞具有体外单克隆生长、自我更新能力以及体内移植成瘤性。

Objective To explore the biological characteristics of side population cells from human bladder cancer cell line HTB-1. Methods The content of SP was determined in cultured HTB-1 cells and after the in vitro amplification.SP and NSP cells were sorted out from human bladder cancer cell line HTB-1.The colony-forming abilities of SP and NSP cells were determined by soft agar assay.The self-renewal abilities of SP and NSP were observed and compared by tumor sphere assay.The tumorigenicities of SP and NSP cells were determined and compared by tumor formation assay. Results The percentage of SP was 0.93% in HTB-1 cells.SP generated SP and NSP cells,while NSP only generated NSP cells.The colony-forming ability of SP was higher than that of NSP cells（P0.01）.The growth rate of tumor sphere of SP was higher than that of NSP（P0.01）.At least 1 000 SP cells could initiate tumor in nude mice. Conclusion The results suggest that SP has colony-forming ability from single cells,self-renewal ability and tumorigenicity.