Estrogen augments shear stress-induced signaling and gene expression in osteoblast-like cells via estrogen receptor-mediated expression of beta1-integrin.
Estrogen and mechanical forces are positive regulators for osteoblast proliferation and bone formation. We investigated the synergistic effect of estrogen and flow-induced shear stress on signal transduction and gene expression in human osetoblast-like MG63 cells and primary osteoblasts (HOBs) using activations of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) and expressions of c-fos and cyclooxygenase-2 (I) as readouts. Estrogen (17beta-estradiol, 10 nM) and shear stress (12 dyn/cm(2)) alone induced transient phosphorylations of ERK and p38 MAPK in MG63 cells. Pretreating MG63 cells with 17beta-estradiol for 6 hours before shearing augmented these shear-induced MAPK phosphorylations. Western blot and flow cytometric analyses showed that treating MG63 cells with 17beta-estradiol for 6 hrs induced their beta(1)-integrin expression. This estrogen-induction of beta(1)-integrin was inhibited by pretreating the cells with a specific antagonist of estrogen receptor ICI 182,780. Both 17beta-estradiol and shear stress alone induced c-fos and Cox-2 gene expressions in MG63 cells. Pretreating MG63 cells with 17beta-estradiol for 6 hrs augmented the shear-induced c-fos and Cox-2 expressions. The augmented effects of 17beta-estradiol on shear-induced MAPK phosphorylations and c-fos and Cox-2 expressions were inhibited by pretreating the cells with ICI 182,780 or transfecting the cells with beta(1)-specific small interfering RNA. Similar results on the augmented effect of estrogen on shear-induced signaling and gene expression were obtained with HOBs. Our findings provide insights into the mechanism by which estrogen augments shear stress responsiveness of signal transduction and gene expression in bone cells via estrogen receptor-mediated increases in beta(1)-integrin expression.
Yeh
Chiuan-Ren
CR
Institute of Biomedical Engineering, National Yang-Ming University, Taiwan, ROC.
Chiu
Jeng-Jiann
JJ
Lee
Chih-I
CI
Lee
Pei-Ling
PL
Shih
Yu-Tsung
YT
Sun
Jui-Sheng
JS
Chien
Shu
S
Cheng
Cheng-Kung
CK
D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D004967:Estrogens / Q000494:pharmacology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism*; D010006:Osteoblasts / Q000187:drug effects*; D011960:Receptors, Estrogen / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033081:Shear Strength; D015398:Signal Transduction / Q000187:drug effects
D004967:Estrogens; D019012:Integrin beta1; D011960:Receptors, Estrogen; D048049:Extracellular Signal-Regulated MAP Kinases