REVERSE TRANSCRIPTASE-POLYMERASE CHAIN-REACTION (RT-PCR) - A SENSITIVE METHOD TO EXAMINE BASIC FIBROBLAST GROWTH FACTOR-INDUCED EXPRESSION OF THE EARLY GROWTH-RESPONSE GENE-1 (EGR-1) IN HUMAN UMBILICAL ARTERIAL ENDOTHELIAL-CELLS

被引：32

作者：

KO, Y ^{[1
]}

TOTZKE, G ^{[1
]}

SCHIERMEYER, B ^{[1
]}

ZEITLER, H ^{[1
]}

SCHMITZ, U ^{[1
]}

VETTER, H ^{[1
]}

SACHINIDIS, A ^{[1
]}

机构：

[1] UNIV BONN, MED POLIKLIN, D-53111 BONN, GERMANY

来源：

MOLECULAR AND CELLULAR PROBES | 1995年 / 9卷 / 04期

关键词：

EARLY GROWTH RESPONSE GENE; FIBROBLAST GROWTH FACTOR; POLYMERASE CHAIN REACTION; REVERSE TRANSCRIPTASE;

D O I：

10.1016/S0890-8508(95)90070-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Immediate-early genes are expressed upon growth and differentiation in a large variety of cells and species. In the present study we investigated the effect of basic fibroblast growth factor (bFGF) on early growth response gene-1 (egr-1)-mRNA expression in human umbilical arterial endothelial cells (HUAEC). The detection of this gene in HUAEC was performed by Northern blotting and by reverse transcriptase-polymerase chain reaction (RT-PCR). For RT-PCR specific primers for egr-1 and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were constructed and PCR conditions were optimized. bFGF induced a time- and concentration-dependent increase of egr-1 expression. Maximal expression occurred within 30 min of stimulation with bFGF at a concentration of 50-100 ng ml(-1). RT-PCR gave highly reproducible and specific results. The comparison of both methods showed comparable results but a higher sensitivity for RT-PCR in detecting the egr-1 mRNA. RT-PCR is an excellent method for detecting the expression of egr-1 mRNA in HUAEC. (C) 1995 Academic Press Limited

引用

页码：215 / 222

页数：8

共 17 条

[1] IDENTIFICATION AND CHARACTERIZATION OF THE EGR-1 GENE-PRODUCT, A DNA-BINDING ZINC FINGER PROTEIN-INDUCED BY DIFFERENTIATION AND GROWTH SIGNALS [J].