Your browser doesn't support javascript.
loading
The preparation of endostatin protein and the measurement of its biologic activity / 国际眼科杂志(Guoji Yanke Zazhi)
International Eye Science ; (12): 841-846, 2005.
Article Dans Chinois | WPRIM | ID: wpr-641781
ABSTRACT
·

AIM:

To investigate the preparation of endostatin protein and its biologic activity on vascular endothelial cell

METHODS:

pBlast-hEndostatin and pBlast-Mcs were identified by digesting with Nhe Ⅰ and Sal Ⅰ, by PCR reaction, by sequencing, and by Alignments of PCR products with gene bank using NCBIBLAST software. The identified pBlast-hEndostatin as well as pBlast-Mcs were then purified with QIAGEN Endofree plasmid maxi kit.The purified plasmids transfected human fibroblasts. The expression of endostatin was detected by RT-PCR, Westem-Blot and immunohistochemistry. The endostatin prorein produced by transfected fibroblasts was purified by ultrafiltration and affinity chromatography. The inhibitory action of endostatin on human umbilical vein endothelium was measured by MTT assay.·

RESULTS:

pBlast-hEndostatin was found to contain human endostatin gene. Endostatin protein was produced by transfected fibroblasts. The inhibitory ratio of 2.5,5,10,20,40,80mg/L endostatin on human umbilical vein endothelium for 48h were 8.5%,13.1%,27.7%,38.1%,56.7%,63.8% respectively. IC50 value was 34.5mg/L.No inhibition action was found on fibroblasts

CONCLUSIONS:

Endostatin protein can be produced by the transfected fibroblasts. The produced endostatin has inhibitory action on human umbilical vein endothelium and has no inhibition action on fibroblasts.

Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) langue: Chinois Texte intégral: International Eye Science Année: 2005 Type: Article

Documents relatifs à ce sujet

MEDLINE

...
LILACS

LIS

Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) langue: Chinois Texte intégral: International Eye Science Année: 2005 Type: Article