Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone.
Human bone marrow stromal cells (hBMSC) are pluripotent cells that have the ability to differentiate into bone, cartilage, hematopoietic-supportive stroma, and adipocytes in a process modulated by dexamethasone (DEX). To characterize changes in hBMSC in response to DEX, we carried out differential display experiments using hBMSC cultured for 1 week in the presence or absence of 10(-8) M DEX. When RNA from these cells was used for differential display, numerous cDNA bands were identified that were up-regulated and down-regulated by DEX. The cDNA bands were reamplified by PCR and directly used to screen an hBMSC cDNA library. Seven clones were isolated and characterized by DNA sequencing and found to encode the following genes: transforming growth factor-beta-induced gene product ((beta)ig-h3), calphobindin II, cytosolic thyroid-binding protein, 22-kDA smooth muscle protein (SM22), and the extracellular matrix proteins osteonectin/SPARC, type III collagen, and fibronectin. To confirm that these genes were regulated by DEX, the cells were treated continuously with this hormone for periods ranging from 2 to 30 days, and steady-state mRNA levels were measured by Northern blot analysis. All genes showed some level of regulation by DEX. The most profound regulation by DEX was observed in the (beta)ig-h3 gene, which showed a relative 10-fold decrease in mRNA levels after 6 days of treatment. Interestingly, (beta)ig-h3 expression was not altered by DEX in fibroblasts from other human tissues, including thymus stromal fibroblasts, spleen stromal fibroblasts, and foreskin fibroblasts. In summary, differential display of DEX-treated hBMSC revealed unique patterns of gene expression and has provided new information about phenotypic changes that accompany the differentiation of hBMSC toward osteogenesis. J. Cell. Biochem. 76:231-243, 1999. Published 1999 Wiley-Liss, Inc.
Dieudonné
S C
SC
Craniofacial and Skeletal Diseases Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
Kerr
J M
JM
Xu
T
T
Sommer
B
B
DeRubeis
A R
AR
Kuznetsov
S A
SA
Kim
I S
IS
Gehron Robey
P
P
Young
M F
MF
D017317:Annexin A6 / Q000235:genetics; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics; D002454:Cell Differentiation; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins; D005353:Fibronectins / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D017403:In Situ Hybridization; D008565:Membrane Proteins / Q000235:genetics; D008840:Microfilament Proteins; D009124:Muscle Proteins / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics; D015676:Osteonectin / Q000235:genetics; D011347:Procollagen / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013963:Thyroid Hormones; D016212:Transforming Growth Factor beta; D000097810:Thyroid Hormone-Binding Proteins; D000099222:betaIG-H3 Protein
D017317:Annexin A6; D002352:Carrier Proteins; D003907:Dexamethasone; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D009124:Muscle Proteins; D009363:Neoplasm Proteins; D015676:Osteonectin; D011347:Procollagen; D012333:RNA, Messenger; D013963:Thyroid Hormones; D016212:Transforming Growth Factor beta; D000097810:Thyroid Hormone-Binding Proteins; D000099222:betaIG-H3 Protein; C058992:Tagln protein, mouse; C051615:transgelin