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CD133 mRNA的表达,流式细胞术检测CD44+CD133+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CD133 mRNA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+ CD133+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.%Objective To isolate and enrich prostate cancer stem cells from PC-3 human prostate cancer cells. Methods PC-3 cells were suspension-cultured in vitro 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 CD44 mRNA and CD133 mRNA in the sphere-forming cells, which had been identified as specific markers of prostate cancer stem 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+ cells and SP cells in the sphere-forming cells were higher significantly than those of the adherent cells(13. 94% vs. 0. 77% and 3.10% 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.
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