Lentiviral-mediated RNA interference against TGF-beta receptor type II in renal epithelial and fibroblast cell populations in vitro demonstrates regulated renal fibrogenesis that is more efficient than a nonlentiviral vector

Yang, Tao, Zhang, Bing, Pat, Betty K., Wei, Ming Q. and Gobe, Glenda C. (2010) Lentiviral-mediated RNA interference against TGF-beta receptor type II in renal epithelial and fibroblast cell populations in vitro demonstrates regulated renal fibrogenesis that is more efficient than a nonlentiviral vector. Journal of Biomedicine and Biotechnology, 2010 859240-1-859240-12. doi:10.1155/2010/859240


Author Yang, Tao
Zhang, Bing
Pat, Betty K.
Wei, Ming Q.
Gobe, Glenda C.
Title Lentiviral-mediated RNA interference against TGF-beta receptor type II in renal epithelial and fibroblast cell populations in vitro demonstrates regulated renal fibrogenesis that is more efficient than a nonlentiviral vector
Journal name Journal of Biomedicine and Biotechnology   Check publisher's open access policy
ISSN 1110-7243
1110-7251
2314-6133
2314-6141
Publication date 2010
Sub-type Article (original research)
DOI 10.1155/2010/859240
Open Access Status DOI
Volume 2010
Start page 859240-1
End page 859240-12
Total pages 12
Place of publication New York, NY, United States
Publisher Hindawi Publishing Corporation
Collection year 2011
Language eng
Formatted abstract
Background. Lentiviral constructs reportedly can integrate into the genome of non-dividing, terminally differentiated cells and dividing cells, for long-term gene expression. This investigation tested whether a third generation lentiviral-mediated small interfering RNA (siRNA) delivered into renal epithelial and fibroblast cells against type II transforming growth factor-beta receptor (siRNA-TBRII) could better attenuate renal fibrogenesis in comparison with a non-lentiviral construct.
Methods. HIV-derived lentiviral and non-lentiviral constructs were used to transfect cells with siRNA-TBRII or siRNA-EGFP control. Human embryonic kidney (HEK-293T), renal epithelial cells (NRK-52E) and renal fibroblasts (NRK-49F) were transfected and gene silencing quantified (fluorescence microscopy, Western blotting, fluorescence-activated cell sorting). Renal fibrogenesis was assessed using extracellular matrix protein synthesis (fibronectin and collagen-III; Western immunoblot), and -smooth muscle actin (α-SMA) was analysed as a marker of fibroblast activation and epithelial-to-mesenchymal transdifferentiation (EMT).
Results. Lentiviral-mediated siRNA-TBRII significantly suppressed TBRII expression in all cell lines, and also significantly suppressed renal fibrogenesis. In comparison with the non-lentiviral construct, lentiviral-mediated siRNA-TBRII produced stronger and more persistent inhibition of collagen-III in NRK-49F cells, fibronectin in all renal cell lines, and α-SMA in renal epithelial cells.
Conclusions. Lentiviral vector systems against TBRII can be delivered into renal cells to efficiently limit renal fibrogenesis by sequence-specific gene silencing.
Copyright © 2010 Tao Yang et al.
Keyword Growth-factor-beta
Mammalian-cells
Kidney-disease
Tubulointerstitial fibrosis
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Article # 859240

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2011 Collection
School of Medicine Publications
 
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Created: Sun, 27 Mar 2011, 00:07:53 EST