人骨髓间充质干细胞条件培养液对前列腺癌PC-3细胞生长的影响

Effect of human bone marrow mesenchymal stem cells conditioned medium on cell growth of prostate cancer cell line PC-3

目的:探讨人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,MSCs)条件培养液(conditioned medium,CM)对前列腺癌PC-3细胞生长的影响。方法:从健康成人骨髓中分离、培养并扩增MSCs。第3代(P3)MSCs融合达80%时更换培养液,继续培养24h后收集上清液,即MSCs-CM。PC-3细胞分别在常规培养液(对照组)和含50%MSCs-CM的培养液(CM处理组)中培养。使用透射电子显微镜观察细胞超微结构,采用WST-8法检测细胞增殖活性,采用FCM法分析细胞周期变化。结果:培养第4天时,透射电子显微镜观察发现,与对照组相比,CM处理组细胞核质比更小,细胞质中细胞器更为丰富。培养第1、3、5和7天时,WST-8法检测显示,2组间除第1天的吸光度(D)值无统计学差异(P>0.05)外,其余3天实验组的D值均高于对照组,差异有统计学意义(P<0.01)。培养第4天时,FCM法检测显示,与对照组相比,CM处理组处于G0/G1期的细胞比例降低[(56.200±0.78)%vs(68.535±1.21)%],同时处于S+G2/M期的细胞比例升高[(43.80±0.77)%vs(31.46±1.20)%],差异有统计学意义(P<0.01)。结论:MSCs-CM可通过加快G1/S细胞周期转换而促进PC-3细胞增殖。

Obiective： To investigate the effect of human bone marrow mesenchymal stem cells （MSCs） conditioned medium （CM） on cell growth of prostate cancer cell line PC-3. Methods= MSCs were isolated and culture-expanded by using density gradient centrifugation from a healthy adult. When the passage 3 （P3） culture reached 80% confluence, the medium was changed to fresh medium. After 24-hour incubation, the supernatant （MSCs-CM） was collected. Then the PC-3 cells were incubated in fresh medium alone （as the control group） or in the medium supplemented with 50% MSCs-CM （as the CM group）. The ultrastructure of PC-3 cells was observed under a transmission electron microscope. The cell viability was measured by WST-8 assay, and the cell cycle was analyzed by flow cytometry （FCM）. ResulCs= On day 4, the nucleus to cytoplasm ratio was lower and the number of organelles was much more in PC-3 cells in the CM group than those in the control group. The WST-8 assay showed that the absorbance values （on day 1, 3, 5 and 7） of the CM group were significantly higher than those in the control group （P〈0.01）, except for the value on day 1 （P〉0.05）. On day 4, the percentage of cells in G0/G1 phase was decreased while which in S＋G2/M phase was increased in the CM group compared with those in the control group （56.20%_＋ 0.78% vs 68.54%±1.21%, P〈0.01; 43.80%＋0.77% vs 31.46%＋1.20%, P〈0.01）. Conclusion： MSCs-CM can promote the proliferation of PC-3 cells through accelerating G/to S phase transition.