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Expression and induction of CYP4F subfamily in human leukocytes and HL60 cells
Yasushi Kikuta1, Yoshiaki Yamashita, Soichiro Kashiwagi
1Department of Applied Biological Science, Faculty of Life Science and Biotechnology, Fukuyama University, Gakuencho-1, Fukuyama, Hiroshima 729-0292, Japan. kikuta@fubac.fukuyama-u.ac.jp
Abstract:
We investigated the expression of the CYP4F subfamily in human leukocytes and HL60 cells. Enzymatic activity assay, immunocytochemical staining, and reverse transcription-polymerase chain reaction (RT-PCR) analysis of human leukocytes showed that polymorphonuclear leukocytes (PMNs) expressed CYP4F3B and CYP4F12 in addition to CYP4F3. Transcription start site of CYP4F3B mRNA in the leukocytes was identical to that of CYP4F3 mRNA. The HL60 cells, which were differentiated into PMN-like shapes by treatment with all-trans-retinoic acid (RA), also expressed CYP4F3, CYP4F3B and CYP4F12. CYP4F3 was expressed in one third of the peripheral monocytes, which omega-hydroxylated leukotriene B(4) (LTB(4)) at a rate 11 times lower than that of PMN. The cells that were differentiated into a form similar to monocytes/macrophages in shape by treatment with 12-myristate 13-acetate expressed mRNA for CYP4F3 and CYP4F3B. Promoter analysis of the CYP4F3 gene demonstrated that a region (-174/-90) of this gene was important for its promoter activity in the HL60 cells. This is the first report on the distribution of different CYP4F isoforms in leukocytes and their induction in HL60 cells.
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