Measurement of Interleukin-1 Stimulated Constitutive Prostaglandin G/H Synthase (Cyclooxygenase) mRNA Levels in Osteoblastic MC3T3-E1 Cells Using Competitive Reverse Transcriptase Polymerase Chain Reaction
- 1 January 1994
- journal article
- Published by Taylor & Francis in Endocrine Research
- Vol. 20 (3) , 219-233
- https://doi.org/10.1080/07435809409035860
Abstract
To assess regulation of constitutive prostaglandin G/H synthase (PGHS-1) by interleukin-1 (IL-1) in osteoblastic MC3T3-E1 cells, we compared analysis by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) with Northern blot analysis. Using RT-PCR, IL-1 increased PGHS-1 mRNA levels by 1.84 +/- 0.10 or 2.07 +/- 0.17, depending on the method of calculation. Using Northern blot analysis, the effect of IL-1 on PGHS-1 mRNA levels was more variable, and the variability was increased by normalization of PGHS-1 mRNA levels to the housekeeping genes, beta-actin and glyceraldehyde phosphate dehydrogenase (GAPDH), because their mRNA levels were also regulated by IL-1. We conclude that competitive RT-PCR is a reproducible and accurate method for studying small changes in mRNA levels.Keywords
This publication has 12 references indexed in Scilit:
- Serum and Glucocorticoid Regulation of Gene Transcription and Expression of the Prostaglandin H Synthase-1 and Prostaglandin H Synthase-2 IsozymesArchives of Biochemistry and Biophysics, 1993
- Evidence for Two Antigenically Distinct Molecular Weight Variants of Prostaglandin HSynthase in the Rat OvaryMolecular Endocrinology, 1991
- Mechanism of Regulation of Prostaglandin Production by Parathyroid Hormone, Interleukin-1, and Cortisol in Cultured Mouse Parietal Bones*Endocrinology, 1991
- Expression of a mitogen-responsive gene encoding prostaglandin synthase is regulated by mRNA splicing.Proceedings of the National Academy of Sciences, 1991
- Regulation of Prostaglandin Endoperoxide Synthase Gene Expression in Cultured Rat Mesangial Cells: Induction by Serum via a Protein Kinase-C-Dependent MechanismMolecular Endocrinology, 1991
- Analysis of cytokine mRNA and DNA: detection and quantitation by competitive polymerase chain reaction.Proceedings of the National Academy of Sciences, 1990
- Quantitation of mRNA by the polymerase chain reaction.Proceedings of the National Academy of Sciences, 1989
- Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel approach by aPCR aidedtranscipttitration assay (PATTY)Nucleic Acids Research, 1989
- Effect of interleukin 1 beta on osteoblastic clone MC3T3-E1 cellsCalcified Tissue International, 1988
- Recombinant human interleukin 1 alpha and beta stimulate mouse osteoblast-like cells (MC3T3-E1) to produce macrophage-colony stimulating activity and prostaglandin E2Biochemical and Biophysical Research Communications, 1986