MEASUREMENT OF INTERLEUKIN-1 STIMULATED CONSTITUTIVE PROSTAGLANDIN G/H SYNTHASE (CYCLOOXYGENASE) MESSENGER-RNA LEVELS IN OSTEOBLASTIC MC3T3-E1 CELLS USING COMPETITIVE REVERSE-TRANSCRIPTASE POLYMERASE CHAIN-REACTION

被引：14

作者：

KAWAGUCHI, H ^{[1
]}

YAVARI, R ^{[1
]}

STOVER, ML ^{[1
]}

ROWE, DW ^{[1
]}

RAISZ, LG ^{[1
]}

PILBEAM, CC ^{[1
]}

机构：

[1] UNIV CONNECTICUT,CTR HLTH,DEPT PEDIAT,FARMINGTON,CT 06030

来源：

ENDOCRINE RESEARCH | 1994年 / 20卷 / 03期

关键词：

D O I：

10.1080/07435809409035860

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

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 oh 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 level's 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.

引用

页码：219 / 233

页数：15

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