omega-hydroxylation activity toward leukotriene B(4) and polyunsaturated fatty acids in the human hepatoblastoma cell line, HepG2, and human lung adenocarcinoma cell line, A549.
The addition of glucose to the culture medium of HepG2 or A549 cells for 22 h caused a dose-dependent increase in leukotriene B(4) omega-hydroxylation activity in the homogenate. The addition of genistein to the culture medium of HepG2 or A549 cells for 22 h caused a dose-dependent decrease in the activity, although the number of living cells was not influenced by the addition of genistein. The inhibition by genistein was reversed by removal of genistein from the culture medium in 22 h. The specific leukotriene B(4) omega-hydroxylation activity was high in the nuclear envelope fraction of HepG2 or A549 cells, and a large portion of the activity was concentrated in the nuclear envelope fraction. In the nuclear envelope fraction, leukotriene B(4) omega-hydroxylation activity was accompanied by high polyunsaturated fatty acid omega-hydroxylation activity. The apparent K(m) values for arachidonic acid and leukotriene B(4) in the fractions of HepG2 or A549 cells were 25 and 50 microM, or 22 and 66 microM, respectively. The V(max) values were 222 and 104 pmol/min/mg protein, or 175 and 370 pmol/min/mg protein, respectively. NADPH-dependent omega-hydroxylation of LTB(4) in the nuclear envelope fraction of HepG2 or A549 cells was strongly inhibited by metyrapone and CO. The expression of cytochrome P450 4F2 mRNAs was detected in HepG2 and A549 cells, and thus the arachidonic acid and leukotriene B(4) omega-hydroxylation activities in the nuclear envelope fractions of HepG2 and A549 cells are likely due to cytochrome P450 4F2.
Yamane
M
M
Department of Biochemistry, Tokyo Medical University, Shinjuku-ku, Tokyo 160, Japan. m-yamane@tokyomed.ac.jp
Abe
A
A
D000230:Adenocarcinoma / Q000378:metabolism*; D003470:Culture Media; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D019833:Genistein / Q000494:pharmacology; D018197:Hepatoblastoma / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D007975:Leukotriene B4 / Q000378:metabolism*; D008175:Lung Neoplasms / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014638:Vanadates / Q000494:pharmacology
D003470:Culture Media; D005231:Fatty Acids, Unsaturated; D007975:Leukotriene B4; D014638:Vanadates; D019833:Genistein