Gene expression profiling of connective tissue growth factor (CTGF) stimulated primary human tenon fibroblasts reveals an inflammatory and wound healing response in vitro.
OBJECTIVE
The biologic relevance of human connective tissue growth factor (hCTGF) for primary human tenon fibroblasts (HTFs) was investigated by RNA expression profiling using affymetrix(TM) oligonucleotide array technology to identify genes that are regulated by hCTGF.


METHODS
Recombinant hCTGF was expressed in HEK293T cells and purified by affinity and gel chromatography. Specificity and biologic activity of hCTGF was confirmed by biosensor interaction analysis and proliferation assays. For RNA expression profiling HTFs were stimulated with hCTGF for 48h and analyzed using affymetrix(TM) oligonucleotide array technology. Results were validated by real time RT-PCR.


RESULTS
hCTGF induces various groups of genes responsible for a wound healing and inflammatory response in HTFs. A new subset of CTGF inducible inflammatory genes was discovered (e.g., chemokine [C-X-C motif] ligand 1 [CXCL1], chemokine [C-X-C motif] ligand 6 [CXCL6], interleukin 6 [IL6], and interleukin 8 [IL8]). We also identified genes that can transmit the known biologic functions initiated by CTGF such as proliferation and extracellular matrix remodelling. Of special interest is a group of genes, e.g., osteoglycin (OGN) and osteomodulin (OMD), which are known to play a key role in osteoblast biology.


CONCLUSIONS
This study specifies the important role of hCTGF for primary tenon fibroblast function. The RNA expression profile yields new insights into the relevance of hCTGF in influencing biologic processes like wound healing, inflammation, proliferation, and extracellular matrix remodelling in vitro via transcriptional regulation of specific genes. The results suggest that CTGF potentially acts as a modulating factor in inflammatory and wound healing response in fibroblasts of the human eye.
Universitätsklinikum Würzburg, Abteilung für Molekulare Innere Medizin, Würzburg, Germany. axel.seher@biozentrum.uni-wuerzburg.de
Axel
A
Seher
Joachim
J
Nickel
Thomas D
TD
Mueller
Susanne
S
Kneitz
Susanne
S
Gebhardt
Tobias Meyer
TM
ter Vehn
Guenther
G
Schlunck
Walter
W
Sebald
D054360:Chemokine CXCL1 / Q000096:biosynthesis; D054427:Chemokine CXCL6 / Q000096:biosynthesis; D055513:Connective Tissue Growth Factor / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D005347:Fibroblasts / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D066298:In Vitro Techniques; D007249:Inflammation; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D016209:Interleukin-8 / Q000096:biosynthesis; D011509:Proteoglycans / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014945:Wound Healing
D054360:Chemokine CXCL1; D054427:Chemokine CXCL6; D016326:Extracellular Matrix Proteins; D036341:Intercellular Signaling Peptides and Proteins; D015850:Interleukin-6; D016209:Interleukin-8; C063488:OGN protein, human; D011509:Proteoglycans; D011994:Recombinant Proteins; C113360:osteoadherin; D055513:Connective Tissue Growth Factor
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