中文摘要：

目的从人前列腺癌细胞系PC-3中分离前列腺癌类干细胞并进行初步鉴定,为前列腺癌干细胞的进一步研究奠定基础。方法分别用含血清及无血清培养基培养前列腺癌PC-3细胞,流式细胞仪检测两种不同培养条件下前列腺癌类干细胞的比例,细胞免疫荧光鉴定前列腺癌类干细胞,MTT法测定并比较前列腺癌类干细胞与前列腺癌PC-3细胞的增殖能力及倍增时间,观察前列腺癌类干细胞诱导分化过程及其分化后细胞的双重免疫荧光表达。结果少量PC-3细胞能在添加了人表皮生长因子（EGF）、人碱性成纤维生长因子（bFGF）和人白血病抑制因子（LIF）的无血清培养基中存活并形成悬浮细胞球团,无血清培养基中前列腺癌类干细胞比例（1.43%）显著高于含血清培养基（0.41%,P〈0.05）。前列腺癌类干细胞的增殖能力（培养7d的细胞数为9.6×105个）强于前列腺癌细胞（培养7d的细胞数为3×105个,P〈0.0005）,倍增时间（21.90h）短于前列腺癌细胞（28.38h,P〈0.0005）,免疫荧光显示前列腺癌类干细胞表达CD133;前列腺癌类干细胞球可在含血清培养基中贴壁分化,且双重荧光检测显示其同时低表达CD44和CD133。结论体外培养的前列腺癌细胞系PC-3中含有少量能表达前列腺癌干细胞表面标志物的前列腺癌类干细胞,并可通过无血清培养基富集这类类肿瘤干细胞。

英文摘要：

Objective To isolate and identify prostate cancer stem-like cells from human prostatic carcinoma cell line PC-3,in order to set a foundation for further investigation of human prostate cancer stem cells （hPCSC）. Methods Prostatic carcinoma cells line PC-3 was cultivated in serum-free medium （SFM） supplemented with growth factor,with the same cells cultivated in serum-supplemented medium （SSM） supplemented with growth factor to serve as the control. The proportion of prostate cancer stem-like cells was detected by flow cytometry,and prostate cancer stem-alike cells were identified by cell immunofluorescence,then the proliferating potentiality and doubling time with prostatic carcinoma cells PC-3 were compared by MTT. The process of inducing differentiation and the dual immunofluorescence expression after differentiation of prostate cancer stem-alike cells were observed. Results In SFM consisting of human epidermal growth factor （EGF）,human basic fibroblast growth factor （bFGF） and human leukaemia inhibitory factor （LIF）,a few PC-3 cells could survive and form suspended cell spheres,and the proportion of prostate cancer stem-alike cells （1.43%） was significantly increased compared with that cultured in SSM （0.41%,P0.05）. The reproductive activity of prostate cancer stem-like cells was stronger than prostatic carcinoma cells （9.6×105 pieces vs 3×105 pieces,P0.000 5）,and the doubling time of the former was shorter than the latter （21.90h vs 28.38h,P0.000 5）. Immunofluorescence showed that prostate cancer stem-like cells expressed CD133. Prostate cancer stem-like cell spheres could differentiate in SSM with low expressim of CD44 and CD133. Conclusions Prostatic carcinoma cell line PC-3 cultured in vitro consists of a small amount of prostate cancer stem-like cells which express surface markers belonging to human prostate cancer stem cells. It is possible to enrich this kind of tumor stem-like cells by SFM.