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.

Y Ko; G Totzke; B Schiermeyer; H Zeitler; U Schmitz; H Vetter; A Sachinidis

doi:10.1016/s0890-8508(95)90070-5

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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.

Y Ko·G Totzke·B Schiermeyer·H Zeitler·U Schmitz·H Vetter·A Sachinidis

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Abstract

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.

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Base Sequence Cells, Cultured DNA Primers DNA-Binding Proteins Early Growth Response Protein 1 Endothelium, Vascular Fibroblast Growth Factor 2 Gene Expression Humans Immediate-Early Proteins Molecular Sequence Data Polymerase Chain Reaction RNA, Messenger Transcription Factors Journal Article Research Support, Non-U.S. Gov't

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Y Ko, G Totzke, B Schiermeyer, H Zeitler, U Schmitz, H Vetter, A Sachinidis. 1995. 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.. https://doi.org/10.1016/s0890-8508(95)90070-5

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