Abedin Mohammad J, Kashio Yumiko, Seki Masako, Nakamura Kazuhiro, Hirashima Mitsuomi
Department of Immunology and Immunopathology, Kagawa Medical University, Kita-Gun, Japan.
J Leukoc Biol. 2003 May;73(5):650-6. doi: 10.1189/jlb.0402163.
Little is known about the roles of galectins, a family of beta-galactoside-binding lectins, in myeloid cell differentiation. In the present experiments, we used HL-60 cells as a model of myeloid cell differentiation. The HL-60 cells were differentiated into eosinophil-, monocyte-, and neutrophil-like cells by coculture with sodium butyrate under a mild alkaline condition, phorbol 12-myristate 13-acetate, and dimethyl sulfoxide, respectively. Thus, the expression of galectins in HL-60 cells during differentiation into three different lineages was assessed. Reverse transcriptase-polymerase chain reaction analyses revealed that undifferentiated HL-60 cells expressed galectin-1, -3, -8, -9, and -10 (identical to Charcot Leyden crystal) mRNAs, and galectin-2, -4, and -7 were negligible before and after the differentiations. We failed to detect evident changes in the mRNA levels of galectin-1 and -8 during the differentiations. However, during the eosinophilic differentiation, galectin-9 mRNA expression was gradually decreased, whereas galectin-10 mRNA expression was increased. During the course of monocytic differentiation, galectin-9 mRNA expression was down-regulated, whereas galectin-3 mRNA expression was up-regulated. Moreover, only galectin-10 mRNA expression was enhanced in the process of neutrophilic differentiation. These changes in galectin expressions were confirmed by Western blot and flow cytometry analyses. It is thus suggested that changes in the expressions of galectin-3, -9, and -10 are potentially important for myeloid cell differentiation into specific lineages.
半乳糖凝集素是一类β-半乳糖苷结合凝集素,关于其在髓样细胞分化中的作用,目前所知甚少。在本实验中,我们使用HL-60细胞作为髓样细胞分化的模型。HL-60细胞分别在轻度碱性条件下与丁酸钠、佛波酯12-肉豆蔻酸酯13-乙酸酯及二甲亚砜共培养,从而分化为嗜酸性粒细胞样、单核细胞样和中性粒细胞样细胞。因此,我们评估了HL-60细胞在分化为三种不同谱系过程中半乳糖凝集素的表达情况。逆转录聚合酶链反应分析显示,未分化的HL-60细胞表达半乳糖凝集素-1、-3、-8、-9和-10(与夏科-莱登结晶相同)的mRNA,而半乳糖凝集素-2、-4和-7在分化前后表达量极低。我们未能检测到半乳糖凝集素-1和-8在分化过程中mRNA水平的明显变化。然而,在嗜酸性粒细胞分化过程中,半乳糖凝集素-9的mRNA表达逐渐降低,而半乳糖凝集素-10的mRNA表达增加。在单核细胞分化过程中,半乳糖凝集素-9的mRNA表达下调,而半乳糖凝集素-3的mRNA表达上调。此外,在中性粒细胞分化过程中,只有半乳糖凝集素-10的mRNA表达增强。这些半乳糖凝集素表达的变化通过蛋白质免疫印迹和流式细胞术分析得到了证实。因此,提示半乳糖凝集素-3、-9和-10表达的变化对于髓样细胞分化为特定谱系可能具有重要意义。