Role of Cbfa1/Runx2 in the fluid shear stress induction of COX-2 in osteoblasts.
Induction of cyclooxygenase-2 (COX-2) is thought to be important for the anabolic effects of mechanical loading. The transcription factor Cbfa1/Runx2 is essential for osteoblastic differentiation. We examined the role of Cbfa1 in the fluid shear stress (FSS) induction of COX-2 in MC3T3-E1 cells stably transfected with a COX-2 promoter-luciferase reporter. Cells were subjected to FSS for 30 min and returned to static culture (post-FSS). COX-2 mRNA and promoter activity peaked 0.5-1h and 2-3h, respectively, post-FSS. Mutation of the Cbfa1 consensus sequence at -267/-261 bp decreased the FSS fold-induction of luciferase activity by 50%. On electrophoretic mobility shift assay (EMSA), proteins binding to an oligonucleotide spanning the Cbfa1 site were supershifted by specific antibody to Cbfa1. FSS did not increase Cbfa1 binding on EMSA or Cbfa1 mRNA or protein levels. These data suggest that transcriptional activity of Cbfa1, independent of its level of expression, is necessary for maximal FSS induction of COX-2 in osteoblasts.
Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA.
Meenal
M
Mehrotra
Masatomo
M
Saegusa
Olga
O
Voznesensky
Carol
C
Pilbeam
D000818:Animals; D002460:Cell Line; D050659:Core Binding Factor Alpha 1 Subunit / Q000096:biosynthesis / Q000502:physiology*; D051546:Cyclooxygenase 2 / Q000096:biosynthesis*; D024202:Electrophoretic Mobility Shift Assay; D004790:Enzyme Induction; D051379:Mice; D010006:Osteoblasts / Q000201:enzymology*; D013314:Stress, Mechanical
D050659:Core Binding Factor Alpha 1 Subunit; C493716:Runx2 protein, mouse; D051546:Cyclooxygenase 2