Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, via Borsari 46, Ferrara, Italy.
Exp Hematol. 2011 Mar;39(3):360-74, 374.e1-5. doi: 10.1016/j.exphem.2010.12.001. Epub 2010 Dec 9.
Extracellular adenosine triphosphate (ATP) is a well-recognized mediator of cell-to-cell communication. Here we show ATP effects on bone marrow (BM)-derived human mesenchymal stem cell (hMSCs) functions.
ATP-induced modification of hMSCs gene expression profile was assessed by Affymetrix technology. Clonogenic and migration assays in vitro, as well as xenotransplant experiments in vivo, were performed to evaluate the effects of ATP on hMSCs proliferation and BM homing. Enzyme-linked immunosorbent assays were used to assess hMSCs cytokines production, whereas T-cell cultures demonstrated the immunoregulatory activity of ATP-treated hMSCs.
hMSCs were resistant to the cytotoxic effects of ATP, as demonstrated by the lack of morphological and mitochondrial changes or release of intracellular markers of cell death. Gene expression profiling revealed that ATP-stimulated hMSCs underwent a downregulation of genes involved in cell proliferation, whereas those involved in cell migration were strongly upregulated. The inhibitory activity of ATP on hMSCs proliferation was confirmed by assessing clonogenic stromal progenitors. ATP potentiated the chemotactic response of hMSCs to the chemokine CXCL12, and increased their spontaneous migration. In vivo, the homing capacity of hMSCs to the BM of immunodeficient mice was significantly increased by pretreatment with ATP. Moreover, ATP increased the production of the proinflammatory cytokines interleukin-2, interferon-γ, and interleukin-12p70, while decreasing the anti-inflammatory cytokine interleukin-10, and this finding was associated with the reduced ability of MSCs to inhibit T-cell proliferation.
Our data show that purinergic signaling modulates hMSCs functions and highlights a role for extracellular nucleotides in hMSCs biology.
细胞外三磷酸腺苷（ATP）是细胞间通讯的公认介质。在这里，我们展示了 ATP 对骨髓（BM）来源的人间充质干细胞（hMSCs）功能的影响。
通过 Affymetrix 技术评估 ATP 诱导的 hMSCs 基因表达谱的改变。体外集落形成和迁移实验以及体内异种移植实验用于评估 ATP 对 hMSCs 增殖和 BM 归巢的影响。酶联免疫吸附测定用于评估 hMSCs 细胞因子的产生，而 T 细胞培养则证明了 ATP 处理的 hMSCs 的免疫调节活性。
hMSCs 对 ATP 的细胞毒性作用具有抗性，这表现在缺乏形态和线粒体变化或细胞死亡的细胞内标志物的释放。基因表达谱分析显示，ATP 刺激的 hMSCs 下调了参与细胞增殖的基因，而强烈上调了参与细胞迁移的基因。通过评估集落形成基质祖细胞，证实了 ATP 对 hMSCs 增殖的抑制活性。ATP 增强了 hMSCs 对趋化因子 CXCL12 的趋化反应，并增加了其自发迁移。在体内，hMSCs 对免疫缺陷小鼠 BM 的归巢能力通过用 ATP 预处理显著增加。此外，ATP 增加了促炎细胞因子白细胞介素-2、干扰素-γ和白细胞介素-12p70 的产生，同时降低了抗炎细胞因子白细胞介素-10 的产生，这与 MSCs 抑制 T 细胞增殖的能力降低有关。
我们的数据表明嘌呤能信号转导调节 hMSCs 的功能，并强调细胞外核苷酸在 hMSCs 生物学中的作用。
