当前位置： SCI文献检索 > OSTEOARTHRITIS AND CARTILAGE期刊下所有文献 > Adenoviral overexpression of Smad-7 and Smad-6 differentially regulates TGF-beta-mediated chondrocyte proliferation and proteoglycan synthesis.

Adenoviral overexpression of Smad-7 and Smad-6 differentially regulates TGF-beta-mediated chondrocyte proliferation and proteoglycan synthesis.

Abstract：

OBJECTIVE:To assess if various biological responses to transforming growth factor-beta (TGF-beta) in chondrocytes are differentially regulated by Smad-6 and Smad-7. DESIGN:Adenoviral overexpression of Smad-6 or -7 mRNA in a chondrocyte cell line was determined via semi-quantitative RT-PCR and protein overexpression was studied by immunocytochemistry. Furthermore, the effect of Smad-6 and -7 overexpression on TGF-beta-induced PAI-1 and aggrecan mRNA upregulation was studied via quantitative RT-PCR. The effect of Smad-6 and -7 overexpression on TGF-beta-induced chondrocyte proliferation was studied via DNA quantification, whereas TGF-beta-induced proteoglycan (PG) synthesis was studied by 35S-sulfate incorporation. RESULTS:Adenoviral transfection of chondrocytes with Smad-6 and -7 resulted in strong upregulation of Smad-6 and -7 mRNA levels, respectively. Immunocytochemistry showed overexpression of Smad-6 and -7 proteins in both the nucleus and cytoplasm. Smad-6 overexpression significantly inhibited TGF-beta-stimulated chondrocyte proliferation, although proliferation was not completely abolished. Smad-7 overexpression, however, completely antagonized the TGF-beta effect on proliferation. Smad-6 overexpression had no effect on TGF-beta-induced PAI-1 expression, while overexpression of Smad-7 completely blocked this TGF-beta effect. Additionally, overexpression of Smad-7, but not Smad-6, totally prevented TGF-beta-induced PG synthesis on the mRNA and protein levels. CONCLUSIONS:Adenoviral transfection of chondrocytes with Smad-6 and -7 resulted in strong upregulation of Smad-6 and -7 mRNA and protein levels. Furthermore, overexpression of Smad-7 in chondrocytes totally inhibited important TGF-beta-mediated biological responses such as proliferation and PG synthesis, while overexpressed Smad-6 had no or only a partial inhibitory effect on TGF-beta activity. We conclude that in chondrocytes distinct TGF-beta activities are differentially regulated by Smad-6 and Smad-7.

journal_name

Osteoarthritis Cartilage

journal_title

Osteoarthritis and cartilage

authors

Scharstuhl A,Diepens R,Lensen J,Vitters E,van Beuningen H,van der Kraan P,van den Berg W

doi

10.1016/s1063-4584(03)00165-1

keywords:

subject

Has Abstract

pub_date

2003-11-01 00:00:00

pages

773-82

issue

11

eissn

1063-4584

issn

1522-9653

pii

S1063458403001651

journal_volume

11

pub_type

杂志文章

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