Extracellular Calcium Is a Potent Inducer of Cyclo-oxygenase-2 in Murine Osteoblasts Through an ERK Signaling Pathway

Abstract

COX-2 mRNA and protein expression were induced by 3-40 mM of [Ca2+]e. [Ca2+]e (5 mM) induced COX-2 mRNA within 30 minutes; levels peaked at 6-9 h and remained elevated at 24 h. Cumulative medium PGE2 was increased at 3 h, with levels rising to 30 nM at 24 h. PGE2 production in POBs from mice with only COX-1 gene expression was 1/40th of that in POBs from mice with both COX-1 and COX-2 gene expression. [Ca2+]e increased alkaline phosphatase activity and osteocalcin mRNA, and this increase was blocked by inhibiting PGE2 production. [Ca2+]e stimulation of COX-2 promoter activity correlated with the induction of COX-2 mRNA expression. [Ca2+]e induced rapid and transient phosphorylation of extracellular signal-regulated kinase (ERK) in POBs, which peaked at 5-10 minutes. Inhibition of ERK phosphorylation with the specific inhibitors, PD-98059 and U-0126, decreased the [Ca2+]e induction of both COX-2 mRNA and luciferase activity by 70-80%. Although less effective than [Ca2+]e, strontium [Sr2+]e also induced COX-2 mRNA and promoter activity in POBs through an ERK signaling pathway. We conclude that [Ca2+]e is a potent transcriptional inducer of COX-2 expression and PGE2 production in osteoblasts through an ERK signaling pathway.