Inhibition of fibroblast-induced angiogenic phenotype of cultured endothelial cells by the overexpression of tissue inhibitor of metalloproteinase (TIMP)-3.
In this study, we examined the effect of overexpression of tissue inhibitor of metalloproteinase (TIMP)-3 on the angiogenic phenotype expressed by vascular endothelial cells (ECs). ECs were infected with a recombinant adenovirus carrying the TIMP-3 gene at various multiplicities of infection, and TIMP-3 expression by transfected cells was confirmed by Western blotting and reverse zymography. At transfection doses of 6.25, 12.5, 25, 50 and 100 multiplicity of infection, EC migration was reduced to 66, 45, 25, 17 and 5%, respectively, of that of the control. At the multiplicity of infection of 20, capillary tube length was reduced by 80% compared to that of the control. Thus, expression of TIMP-3 by ECs effectively inhibited EC migration and tube formation. Overexpression of TIMP-3 by ECs may be considered a gene therapy strategy for the treatment of pathological angiogenesis such as cancer and diabetic retinopathy.
Ma
David Hui-Kang
DH
Department of Ophthalmology, Chang-Gung Memorial Hospital, Chang-Gung University, Kweishan, Taoyuan, Taiwan.
Chen
Jun-I
JI
Zhang
Fen
F
Hwang
David G
DG
Chen
Jan-Kan
JK
D001483:Base Sequence; D002465:Cell Movement; D002478:Cells, Cultured; D018920:Coculture Techniques; D017931:DNA Primers; D004730:Endothelium, Vascular / Q000166:cytology* / Q000201:enzymology / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D018919:Neovascularization, Physiologic / Q000235:genetics / Q000502:physiology*; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics*; D014471:Umbilical Veins
D017931:DNA Primers; D019717:Tissue Inhibitor of Metalloproteinase-3