中文摘要：

目的从PC-3人前列腺癌细胞中分离并富集前列腺癌干细胞。方法体外无血清悬浮培养PC-3细胞，通过传代更新、诱导分化验证PC-3悬浮成球细胞具有强增殖性、自我更新及分化潜能，实时荧光定量核酸扩增检测系统检测CD44和CDl33mRNA的表达，流式细胞术检测CD44＋CDl33＋细胞和侧群（SP）细胞比例。结果PC-3成球细胞可以在无血清培养液（SFM）中生存并形成可以稳定传代的悬浮细胞球。PC-3成球细胞具有自我更新和分化能力，其前列腺癌干细胞标志物CD44和CDl33mRNA的相对表达量分别是PC-3贴壁细胞的41．83倍和10．48倍（P〈0．05）。PC-3悬浮成球细胞中CD44＋CDl33＋细胞比率为13．94％，SP细胞含量为3．10％，均显著高于pC．3贴壁细胞的0．77％和0（P〈0．05）。结论通过无血清悬浮聚球培养可以从PC-3细胞中简便、高效地富集前列腺癌干细胞。

英文摘要：

Objective To isolate and enrich prostate cancer stem cells from PC-3 human prostate cancer cells, Methods PC-3 cells were suspension-cultured in vi（ro with a serum-free medium （SFM）. Then, the floating PC-3 sphere-forming cells were verified to have the abilities of strong proliferation,self-renewal and differentiation potential. The expressions of CD44 mRNA and CD133 mRNA were detected with quantitative RT-PCR（QPCR）. The percentages of CD44＋ CD133＋ cells and side population（SP） cells were detected with flow cytometry. Results The PC-3 sphere-forming cells could survive in SFM and form floating cell spheres, which could propagate stably in vitro. The PC-3 sphere-forming cells possessed the potentials of self-renewal and differentiation. The relative expression quantity of CIM4 mRNA and CD133 mRNA in the sphere-forming cells, which had been identified as specific markers of prostate cancer stern cells,was 41.83 and 10. 48 fold times higher than those of the PC-3 adherent cells, respectively（P〈0. 05）. The percentages of CD44＋ CD133＋ ceils and SP cells in the sphere-forming ceils were higher significantly than those of the adherent cells（13. 94% vs. 0.77% and 3.10M vs. 0）（P〈0. 05）. Conclusion Using suspension culture method with SFM, the prostate cancer stem cells can be enriched conveniently and effectively from PC-3 cells,