Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone

Suzanne C. Dieudonné, Janet M. Kerr, Tianshun Xu, Beatrice Sommer, Anna R. DeRubeis, Sergei A. Kuznetsov, In-San Kim, Pamela Gehron Robey, Marian F. Young

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

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-β-induced gene product (βig-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 βig-h3 gene, which showed a relative 10-fold decrease in mRNA levels after 6 days of treatment. Interestingly, βig-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.

Original languageEnglish
Pages (from-to)231-243
Number of pages13
JournalJournal of cellular biochemistry
Volume76
Issue number2
DOIs
Publication statusPublished - 1999 Dec 1
Externally publishedYes

Fingerprint

Stromal Cells
Dexamethasone
Bone Marrow
Display devices
Mesenchymal Stromal Cells
Bone
Messenger RNA
Fibroblasts
Genes
Immunoglobulin Genes
Annexin A6
Complementary DNA
Osteonectin
Foreskin
Thymus
Collagen Type III
Muscle Proteins
Extracellular Matrix Proteins
Cartilage
Transforming Growth Factors

Keywords

  • βig-h3
  • Dexamethasone
  • Differential display
  • Human bone marrow stroma

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone. / Dieudonné, Suzanne C.; Kerr, Janet M.; Xu, Tianshun; Sommer, Beatrice; DeRubeis, Anna R.; Kuznetsov, Sergei A.; Kim, In-San; Robey, Pamela Gehron; Young, Marian F.

In: Journal of cellular biochemistry, Vol. 76, No. 2, 01.12.1999, p. 231-243.

Research output: Contribution to journalArticle

Dieudonné, Suzanne C. ; Kerr, Janet M. ; Xu, Tianshun ; Sommer, Beatrice ; DeRubeis, Anna R. ; Kuznetsov, Sergei A. ; Kim, In-San ; Robey, Pamela Gehron ; Young, Marian F. / Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone. In: Journal of cellular biochemistry. 1999 ; Vol. 76, No. 2. pp. 231-243.
@article{dbd2da702d1f4469b3a97211dd319f14,
title = "Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone",
abstract = "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-β-induced gene product (βig-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 βig-h3 gene, which showed a relative 10-fold decrease in mRNA levels after 6 days of treatment. Interestingly, βig-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.",
keywords = "βig-h3, Dexamethasone, Differential display, Human bone marrow stroma",
author = "Dieudonn{\'e}, {Suzanne C.} and Kerr, {Janet M.} and Tianshun Xu and Beatrice Sommer and DeRubeis, {Anna R.} and Kuznetsov, {Sergei A.} and In-San Kim and Robey, {Pamela Gehron} and Young, {Marian F.}",
year = "1999",
month = "12",
day = "1",
doi = "10.1002/(SICI)1097-4644(20000201)76:2<231::AID-JCB7>3.0.CO;2-X",
language = "English",
volume = "76",
pages = "231--243",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone

AU - Dieudonné, Suzanne C.

AU - Kerr, Janet M.

AU - Xu, Tianshun

AU - Sommer, Beatrice

AU - DeRubeis, Anna R.

AU - Kuznetsov, Sergei A.

AU - Kim, In-San

AU - Robey, Pamela Gehron

AU - Young, Marian F.

PY - 1999/12/1

Y1 - 1999/12/1

N2 - 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-β-induced gene product (βig-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 βig-h3 gene, which showed a relative 10-fold decrease in mRNA levels after 6 days of treatment. Interestingly, βig-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.

AB - 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-β-induced gene product (βig-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 βig-h3 gene, which showed a relative 10-fold decrease in mRNA levels after 6 days of treatment. Interestingly, βig-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.

KW - βig-h3

KW - Dexamethasone

KW - Differential display

KW - Human bone marrow stroma

UR - http://www.scopus.com/inward/record.url?scp=0033451485&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033451485&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-4644(20000201)76:2<231::AID-JCB7>3.0.CO;2-X

DO - 10.1002/(SICI)1097-4644(20000201)76:2<231::AID-JCB7>3.0.CO;2-X

M3 - Article

C2 - 10618640

AN - SCOPUS:0033451485

VL - 76

SP - 231

EP - 243

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 2

ER -