Characterization of group X phospholipase A(2) as the major enzyme secreted by human keratinocytes and its regulation by the phorbol ester TPA.
HaCaT as well as human primary keratinocytes constitutively expressed mRNA of the human secreted phospholipase A(2) subtype groups X, V, IIA, and IID. A similar expression pattern was also found in human skin biopsies. Protein analysis showed that under serum-free conditions only group X secreted phospholipase A(2) is secreted into cell culture supernatants of HaCaT as well as human primary keratinocytes, whereas the other secreted phospholipases A(2) were not detectable at protein level. HaCaT keratinocytes constitutively released secreted phospholipase A(2) activity into the cell culture supernatant, being reflected by a constant release of fatty acids. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate, which is a potent inducer of inflammation in skin, drastically reduced the mRNA level of group X secreted phospholipase A(2) and other secreted phospholipase A(2) subtypes as well as secreted phospholipase A(2) activity in cell culture supernatants. This suggests that inhibition of secreted phospholipase A(2) expression and activity as well as of fatty acid release by 12-O-tetradecanoylphorbol-13-acetate treatment might be a critical step impairing the integrity of the epidermis during phorbol-ester-induced pathologic processes in skin. The results show that group X secreted phospholipase A(2) represents the major secreted phospholipase A(2) subtype in human keratinocytes and thus may indicate a physiologic role for this enzyme in epidermis in vivo.
Schadow
A
A
Pharmazentrum Frankfurt, University Hospital, Frankfurt/Main, Germany.
Scholz-Pedretti
K
K
Lambeau
G
G
Gelb
M H
MH
Fürstenberger
G
G
Pfeilschifter
J
J
Kaszkin
M
M
D015153:Blotting, Western; D002460:Cell Line / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D015603:Keratinocytes / Q000201:enzymology* / Q000378:metabolism*; D010741:Phospholipases A / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D012636:Secretory Rate / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D005230:Fatty Acids, Nonesterified; D007527:Isoenzymes; D012333:RNA, Messenger; D010741:Phospholipases A; D013755:Tetradecanoylphorbol Acetate