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Developing a system for regulating expression of human hepatocyte growth factor using tetracycline in NRK52E cells.
Ren, Shu-Ting; Yu, Lin-Hua; Zhang, Jian; Han, Shui-Ping; Xu, Chang-Fu.
Afiliação
  • Ren ST; Department of Pathology, School of Medicine, Xi'an Jiaotong University, Xi'an, China.
Urol Int ; 85(2): 228-36, 2010.
Article em En | MEDLINE | ID: mdl-20699604
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 AND METHODS: 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.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Tetraciclina / Fator de Crescimento de Hepatócito / Células Epiteliais / Túbulos Renais Limite: Animals / Humans Idioma: En Ano de publicação: 2010 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Tetraciclina / Fator de Crescimento de Hepatócito / Células Epiteliais / Túbulos Renais Limite: Animals / Humans Idioma: En Ano de publicação: 2010 Tipo de documento: Article