Developing a system for regulating expression of human hepatocyte growth factor using tetracycline in NRK52E cells.
Urol Int
; 85(2): 228-36, 2010.
Article
de En
| MEDLINE
| ID: mdl-20699604
ABSTRACT
INTRODUCTION:
Hepatocyte growth factor (HGF) is a target of gene therapy for renal fibrosis. The aim of this study was to establish a human HGF gene expression system that is regulated by tetracycline (Tet) in normal rat kidney tubular epithelial cells (NRK52E cells). MATERIALS ANDMETHODS:
The plasmids pTet-on, pBI-L-HGF and pTK-Hyg were transfected sequentially into NRK52E cells using Lipofectamine 2000. The expression of HGF gene was measured, and the activity of expressed HGF was detected.RESULTS:
A clone of pBI-L-HGF/NRK52E cells showing strong reaction to doxycycline (Dox) was selected using a luciferase reporter assay system. The expression of both HGF mRNA and protein was significantly higher (both p < 0.01) in the Dox group than that in the control group. Furthermore, the bioactivity of expressed HGF was confirmed in the assay.CONCLUSIONS:
A Tet-regulated human HGF gene expression system in NRK52E cells has been established. This cell line may prove useful for gene therapy against renal fibrosis.
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Tétracycline
/
Facteur de croissance des hépatocytes
/
Cellules épithéliales
/
Tubules rénaux
Limites:
Animals
/
Humans
Langue:
En
Journal:
Urol Int
Année:
2010
Type de document:
Article
Pays d'affiliation:
Chine