Prostaglandin synthase 2 expression in epidermal growth factor-dependent proliferation of mouse keratinocytes

被引:23
作者
Loftin, CD [1 ]
Eling, TE [1 ]
机构
[1] NIEHS,MOLEC BIOPHYS LAB,NIH,RES TRIANGLE PK,NC 27709
关键词
epidermal growth factor; prostaglandins; prostaglandin synthase 2; arachidonic acid; linoleic acid; HODE; keratinocytes; mitogenesis;
D O I
10.1006/abbi.1996.0271
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
BALB/c mouse keratinocytes (BALB/MK) are nontumorigenic epithelial cells which are dependent on mouse epidermal growth factor (EGF) for maintaining proliferation in culture, In BALB/MK the oxygenation of both arachidonic acid and linoleic acid was dependent on EGF. EGF stimulated the formation of prostaglandin E(2) and prostaglandin F-2 alpha from arachidonic acid and 9- and 13-hydroxyoctadecadienoic acid (HODE) from linoleic acid, Analysis of the linoleic acid metabolites determined the ratio of 9-HODE to 13-HODE was approximately 6 to 4, and the 9-HODE was the (R) enantiomer, consistent with metabolism by prostaglandin G/H synthase (PGHS). The formation of these linoleic acid metabolites was sensitive to indomethacin, a PGHS inhibitor. EGF induced the expression of PGHS-2 mRNA after 30 min, which peaked after 1 h, and remained expressed for at least 24 h after the addition of EGF. A less significant increase in the expression of PGHS-1 mRNA occurred 4 h after EGF stimulation, Immunoblot analysis did not detect expression of PGHS-1 protein. However, PGHS-2 protein expression was increased 2 h after EGF exposure and was dependent on EGF. PGHS-2 protein was not transiently expressed as reported with other cell types, but was continually expressed in proliferating cells maintained with EGF at a subconfluent density, Indomethacin significantly attenuated EGF-dependent mitogenesis and cell proliferation. These results suggest that PGHS-2 activity contributes to the proliferative response of BALB/MK to EGF. (C) 1996 Academic Press, Inc.
引用
收藏
页码:419 / 429
页数:11
相关论文
共 51 条
[1]   PROSTAGLANDIN-H SYNTHASE-2 IS INDUCED IN SYRIAN-HAMSTER EMBRYO CELLS IN RESPONSE TO BASIC FIBROBLAST GROWTH-FACTOR [J].
ANGERMANSTEWART, JA ;
ELING, TE ;
GLASGOW, WC .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1995, 318 (02) :378-386
[2]   INVIVO ACTIVATION OF AN DELTA-6 OXYGENASE IN HUMAN SKIN [J].
BAER, AN ;
COSTELLO, PB ;
GREEN, FA .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 180 (01) :98-104
[3]  
BAER AN, 1990, J LIPID RES, V31, P125
[4]   ENZYMATIC-PROPERTIES OF THE 15-LIPOXYGENASE OF HUMAN CULTURED KERATINOCYTES [J].
BURRALL, BA ;
CHEUNG, M ;
CHIU, A ;
GOETZL, EJ .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 1988, 91 (04) :294-297
[5]  
CHEN YNP, 1987, J BIOL CHEM, V262, P16892
[6]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[7]   SELECTIVE-INHIBITION OF GROWTH-RELATED GENE-EXPRESSION IN MURINE KERATINOCYTES BY TRANSFORMING GROWTH FACTOR-BETA [J].
COFFEY, RJ ;
BASCOM, CC ;
SIPES, NJ ;
GRAVESDEAL, R ;
WEISSMAN, BE ;
MOSES, HL .
MOLECULAR AND CELLULAR BIOLOGY, 1988, 8 (08) :3088-3093
[9]   PROSTAGLANDIN ENDOPEROXIDE SYNTHASE - REGULATION OF ENZYME EXPRESSION [J].
DEWITT, DL .
BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1083 (02) :121-134
[10]   SERUM AND GLUCOCORTICOID REGULATION OF GENE-TRANSCRIPTION AND EXPRESSION OF THE PROSTAGLANDIN-H SYNTHASE-1 AND PROSTAGLANDIN-H SYNTHASE-2 ISOZYMES [J].
DEWITT, DL ;
MEADE, EA .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1993, 306 (01) :94-102