Nishihira J, Koyama Y, Mizue Y
Central Research Institute, School of Medicine, Hokkaido University, Sapporo, Japan.
Biochem Mol Biol Int. 1996 Nov;40(5):861-9. doi: 10.1080/15216549600201473.
Cytokines play an important role in inflammation and immunity. In this study, we examined the expression and presence of human macrophage migration inhibitory factor (MIF) in human myelomonocytic leukemia cell line, HL-60 by reverse transcription-polymerase chain reaction (RT-PCR) and immunological methods (Western blot analysis and immunohistochemistry), respectively. The RT-PCR showed that MIF mRNA was constitutively expressed, and the expression was further induced by lipopolysaccharide (LPS) stimulation. The expression was upregulated by LPS at the range of 10 pg/ml to 10 ng/ml; however, it decreased at doses higher than 100 ng/ml. The expression reached the maximum 12 hr after the stimulation, but substantially decreased by 24 hr. Western blot analysis and immunohistochemistry using an anti-human MIF antibody revealed the presence of MIF protein in cytoplasm of the cells. The pathophysiological role of MIF in HL-60 cells has not been fully understood; however, the regulation of MIF mRNA expression by LPS suggests the possibility that the cytokine plays an important role in inflammatory events of leukemia.
细胞因子在炎症和免疫中发挥着重要作用。在本研究中,我们分别通过逆转录-聚合酶链反应(RT-PCR)和免疫学方法(蛋白质印迹分析和免疫组织化学)检测了人巨噬细胞移动抑制因子(MIF)在人骨髓单核细胞白血病细胞系HL-60中的表达及存在情况。RT-PCR结果显示,MIF mRNA呈组成性表达,且脂多糖(LPS)刺激可进一步诱导其表达。LPS在10 pg/ml至10 ng/ml范围内上调该表达;然而,在高于100 ng/ml的剂量下表达下降。刺激后12小时表达达到最大值,但在24小时时大幅下降。使用抗人MIF抗体进行的蛋白质印迹分析和免疫组织化学显示,细胞胞质中存在MIF蛋白。MIF在HL-60细胞中的病理生理作用尚未完全阐明;然而,LPS对MIF mRNA表达的调节提示该细胞因子在白血病炎症事件中发挥重要作用的可能性。