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Identification of macrophage migration inhibitory factor in human leukemia HL-60 cells and its induction by
J Nishihira1, Y Koyama, Y Mizue
1Central Research Institute, School of Medicine, Hokkaido University, Sapporo, Japan.
Insights
Human macrophage migration inhibitory factor (MIF) is constitutively expressed in leukemia cells. Lipopolysaccharide (LPS) stimulation upregulates MIF mRNA and protein, suggesting a role in leukemia inflammation.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Cytokines are crucial mediators of inflammation and immune responses.
- Human macrophage migration inhibitory factor (MIF) is a key cytokine involved in immune regulation.
Purpose of the Study:
- To investigate the expression and presence of human MIF in the HL-60 myelomonocytic leukemia cell line.
- To determine the effect of lipopolysaccharide (LPS) on MIF expression in HL-60 cells.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) for MIF mRNA detection.
- Western blot analysis and immunohistochemistry for MIF protein detection.
- Stimulation of HL-60 cells with varying concentrations of LPS.
Main Results:
- MIF mRNA was constitutively expressed in HL-60 cells.
- LPS stimulation upregulated MIF mRNA expression in a dose-dependent manner (10 pg/ml to 10 ng/ml), with peak expression at 12 hours.
- MIF protein was detected in the cytoplasm of HL-60 cells.
Conclusions:
- MIF is constitutively expressed at both mRNA and protein levels in HL-60 leukemia cells.
- LPS modulates MIF mRNA expression, indicating its potential involvement in inflammatory processes within leukemia.
- Further research is needed to elucidate the specific pathophysiological role of MIF in HL-60 cells.
Abstract:
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.
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