ABSTRACT
1α,25-Dihydroxyvitamin D3 (1α,25(OH)2D3) regulates osteoblasts through genomic and rapid membrane-mediated responses. Here we examined the interaction of protein disulfide isomerase family A, member 3 (Pdia3) and the traditional vitamin D receptor (VDR) in plasma membrane-associated responses to 1α,25(OH)2D3. We found that Pdia3 co-localized with VDR and the caveolae scaffolding protein, caveolin-1 on the surface of MC3T3-E1 osteoblasts. Immunoprecipitation showed that both Pdia3 and VDR interacted with caveolin-1. Pdia3 further interacted with phospholipase A2 activating protein (PLAA), whereas VDR interacted with c-Src. 1α,25(OH)2D3 changed the interactions and transport of the two receptors and rapidly activated phospholipase A2 (PLA2) and c-Src. Silencing either receptor or caveolin-1 inhibited both PLA2 and c-Src, indicating that the two receptors function interdependently. These two receptor dependent rapid responses to 1α,25(OH)2D3 regulated gene expression, proliferation and apoptosis of MC3T3-E1 cells. These data demonstrate the importance of both receptors and caveolin-1 in mediating membrane responses to 1α,25(OH)2D3 and subsequently regulating osteoblast biology.
Subject(s)
Osteoblasts/metabolism , Protein Disulfide-Isomerases/metabolism , Receptors, Calcitriol/metabolism , Vitamin D/analogs & derivatives , 3T3 Cells , Animals , CSK Tyrosine-Protein Kinase , Caveolin 1/metabolism , Cell Membrane/metabolism , Mice , Osteoblasts/cytology , Proteins/metabolism , Signal Transduction , Vitamin D/metabolism , src-Family Kinases/metabolismABSTRACT
Objective:To investigate the expression of osteopontin (OPN) mRNA and protein and its correlation with clinico-pathological features of salivary gland tumors. Methods:OPN mRNA and protein level were detected by fluorescent quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR)and Western blot in 41 cases of malignant salivary gland tumors,21 of benign salivary gland tumors and 10 of normal salivary gland tissues. The relationship between the relative content of OPN mRNA and protein and clinico-pathological features of salivary gland tumors were analyzed. Results:The differences of OPN mRNA and protein expression levels in various tissues (malignant salivary gland tumors,benign alivary gland tumors and normal salivary gland tissues) were statistically significant,and the mRNA expression levels were correlated significantly with node metastasis and TNMstage,the expression levels of OPN protein were correlated significantly with node metastasis and TNMstage(P
ABSTRACT
Objective To explore the effects of Zhu Ling decoction and Alisma orientalis on the osteopotin mRNA expression in the experimental rat model of renal stone. Methods The experimental rat model of renal stone was set up using glyoxylic acid. Osteopontin mRNA expression in the renal tissues in the experimental rat model of renal stone was detected by RT-PCR. Results Glyoxylic acid obviously increased osteopontin mRNA expression in the renal tissues. Zhu Ling decoction and Alisma orientalis remarkably decreased the expression of osteopontin mRNA in the renal tissues. Conclusion Zhu Ling decoction and Alisma orientalis could decrease the osteopotin mRNA expression in the renal tissues in the experimental rat model of renal stone. The results were helpful for prevention and treatment of renal stone.
