Adenovirus vector-mediated gene transduction to chondrocytes: in vitro evaluation of therapeutic efficacy of transforming growth factor-beta 1 and heat shock protein 70 gene transduction.
OBJECTIVE
To investigate the effects of adenovirus vector-mediated gene transduction of E. coli beta-galactosidase (LacZ), transforming growth factor-beta 1 (TGF-beta 1), and heat shock protein 70 (HSP 70) on human chondrocyte-like cell line (HCS-2/8).


METHODS
We examined expression of transduced genes and their expression periods by 5 bromo-4-chloroindolyl-beta-D-galactoside (X-gal) staining. Northern blotting, ELISA, and Western blotting. To assess the influence of TGF-beta 1 and HSP70 gene transduction, the expression of mRNA of type II collagen, proteoglycan core protein, matrix metalloproteinase 3 (MMP3) and tissue inhibitor of matrix metalloproteinase 1 were examined by Northern blotting.


RESULTS
Staining with X-gal indicated that the genes were transduced into a majority of the cells. Expression of the transduced genes in the cells was continued for at least 21 days. Transduction of TGF-beta 1 gene enhanced mRNA expression of type II collagen and proteoglycan core protein, but suppressed MMP3 mRNA expression in the cells. Expression of HSP70 was also high. Enhanced expression of HSP70 elevated mRNA expression of proteoglycan core protein.


CONCLUSIONS
These results indicate adenovirus vector is useful in chondrocyte gene therapy, and it could be an efficient mediator of TGF-beta 1 and HSP70 gene transduction.
Arai
Y
Y
Department of Orthopaedic Surgery, Kyoto Prefectural University, Japan.
Kubo
T
T
Kobayashi
K
K
Takeshita
K
K
Takahashi
K
K
Ikeda
T
T
Imanishi
J
J
Takigawa
M
M
Hirasawa
Y
Y
D000256:Adenoviridae / Q000235:genetics*; D053674:Aggrecans; D015152:Blotting, Northern; D002356:Cartilage / Q000378:metabolism*; D002452:Cell Count; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D016326:Extracellular Matrix Proteins; D015316:Genetic Therapy; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007592:Joint Diseases / Q000628:therapy; D037181:Lectins, C-Type; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis / Q000235:genetics; D011509:Proteoglycans; D012333:RNA, Messenger / Q000096:biosynthesis; D019714:Tissue Inhibitor of Metalloproteinases; D014161:Transduction, Genetic; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D053674:Aggrecans; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D018840:HSP70 Heat-Shock Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; D016212:Transforming Growth Factor beta; D003094:Collagen; D001616:beta-Galactosidase; D019278:Matrix Metalloproteinase 3