Dipartimento di Scienze Biomediche, Università di Sassari, Sassari, Italy.
Cell Prolif. 2013 Jun;46(3):328-47. doi: 10.1111/cpr.12030.
To analyse proliferation, differentiation and apoptosis in THP-1 cells after stimulation with phorbol 12-myristate 13-acetate (PMA) and retinoic acid (RA).
PMA and RA were used in a three-step-procedure: (i) treatment with 6, 30, 60 nm PMA, that induced initial, intermediate and advanced levels of monocyte-macrophage transition, respectively; (ii) recovery in PMA-free medium; (iii) incubation with 4 μm RA. Cultures were characterized cytokinetically (flow cytometry/bromodeoxyuridine uptake) and immunocytochemically (static cytometry) for expression of CD14, CD11b (monocyte-macrophage) and DC-SIGN (dendritic cell: DCs) markers.
Some treatments determined appearance of monocyte/macrophage, dendritic and apoptotic phenotypes, percentages of which were related to PMA dose used in step 1, and dependent on presence/absence of PMA and RA. PMA withdrawal induced dedifferentiation and partial restoration of proliferative activity, specially in 6 and 30 nm PMA-derived cells. Recovery in the presence of serum (fundamental to DC appearance) indicated that depending on differentiation level, cell proliferation and apoptosis were inversely correlated. Treatment with 30 nm PMA induced intermediate levels of monocytic-macrophagic differentiation, with expression of alternative means of differentiation and acquisition of DCs without using cytokines, after PMA withdrawal and RA stimulation.
Our experimental conditions favoured differentiation, dedifferentiation and transdifferentiational pathways, in monocytic THP-1 cells, the balance of which could be related to both cell proliferation and cell death.
分析佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）和维甲酸（RA）刺激 THP-1 细胞后的增殖、分化和凋亡。
采用三步法用 PMA 和 RA 处理：（i）分别用 6、30、60nm PMA 处理，诱导单核细胞-巨噬细胞转化的初始、中间和晚期水平；（ii）在无 PMA 的培养基中恢复；（iii）用 4μm RA 孵育。用细胞动力学（流式细胞术/溴脱氧尿苷摄取）和免疫细胞化学（静态细胞术）对 CD14、CD11b（单核细胞-巨噬细胞）和 DC-SIGN（树突状细胞：DC）标志物的表达进行特征描述。
一些处理方法确定了单核细胞/巨噬细胞、树突状和凋亡表型的出现，其百分比与步骤 1 中使用的 PMA 剂量有关，并且依赖于 PMA 和 RA 的存在/不存在。PMA 撤回诱导去分化和增殖活性的部分恢复，特别是在 6nm 和 30nm PMA 衍生的细胞中。在有血清（对 DC 出现至关重要）的情况下恢复表明，取决于分化水平，细胞增殖和凋亡呈负相关。用 30nm PMA 处理可诱导中间水平的单核细胞-巨噬细胞分化，在 PMA 撤回和 RA 刺激后，通过替代分化方式表达，并获得不使用细胞因子的 DC。
我们的实验条件有利于单核细胞 THP-1 细胞的分化、去分化和转分化途径，其平衡可能与细胞增殖和细胞死亡有关。
