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Developing a system for regulating expression of human hepatocyte growth factor using tetracycline in NRK52E cells
- Source :
- Urologia internationalis. 85(2)
- Publication Year :
- 2010
-
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 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.
- Subjects :
- Epithelial-Mesenchymal Transition
Tetracycline
Urology
medicine.medical_treatment
Genetic enhancement
Transfection
Cell Line
Transforming Growth Factor beta1
Gene expression
medicine
Renal fibrosis
Animals
Humans
RNA, Messenger
business.industry
Hepatocyte Growth Factor
Growth factor
Epithelial Cells
Molecular biology
Rats
Up-Regulation
Human hepatocyte
Kidney Tubules
Gene Expression Regulation
Hepatocyte nuclear factor 4 alpha
Doxorubicin
Cancer research
Hepatocyte growth factor
business
medicine.drug
Subjects
Details
- ISSN :
- 14230399
- Volume :
- 85
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Urologia internationalis
- Accession number :
- edsair.doi.dedup.....1d0b1019c3f6df012befd4e234ad06ac