title: Adenovirus-mediated P311 inhibits TGF-β1-induced epithelial-mesenchymal transition in NRK-52E cells via TGF-β1-Smad-ILK pathway.
abstract: P311, a highly conserved 8-kDa intracellular protein, has been indicated as an important factor in myofibroblast transformation and in the progression of fibrosis. In the present study, we constructed a recombinant adenovirus vector of p311 (called Ad-P311) and transferred it into rat renal proximal tubular epithelial cells (NRK-52E) to explore the effect of P311 on epithelial-mesenchymal transition (EMT) of NRK-52E cells induced by TGF-β1 and to elucidate its underlying mechanism against EMT. After successfully construction of Ad-P311 and transfer into NRK-52E cells, the proliferation and growth of P311-expressing cells was detected by MTT assay. TGF-β1 was used to induce NRK-52E cells and Western blot analysis was used to examine the EMT markers (E-cadherin and α-smooth muscle actin (α-SMA)), signal transducers (p-Smad2/3 and Smad7). Integrin Linked Kinase (ILK) as a key intracellular mediator that controls TGF-β1-induced-EMT was also assayed by Western blot analysis. The results showed that P311 transfection could significantly inhibit the proliferation and growth of TGF-β1 induced NRK-52E cells. The results also showed that TGF-β1 could induce EMT in NRK-52E cells through Smad-ILK signaling pathway with an increase in α-SMA, pSmad2/3 and ILK expression, and a decrease in E-cadherin and Smad7 expression. However, P311 efficiently blocked Smad-ILK pathway activation and attenuated all these EMT changes induced by TGF-β1. These findings suggest that P311 might be involved in the pathogenesis of renal fibrosis by inhibiting the EMT process via TGF-β1-Smad-ILK pathway. P311 might be a novel target for the control of renal fibrosis and the progression of CKD.
authors.affiliation: Department of Traditional Chinese Medicine, Shandong Provincial Hospital affiliated to Shandong University.
authors.forename: Fanghua
authors.initials: F
authors.lastname: Qi
authors.forename: Pingping
authors.initials: P
authors.lastname: Cai
authors.forename: Xiang
authors.initials: X
authors.lastname: Liu
authors.forename: Min
authors.initials: M
authors.lastname: Peng
authors.forename: Guomin
authors.initials: G
authors.lastname: Si
mesh_terms: D000256:Adenoviridae; D000818:Animals; D058750:Epithelial-Mesenchymal Transition; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D009400:Nephrosclerosis / Q000209:etiology* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051381:Rats; D051785:Smad Proteins / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D000099200:Scaffold Protein ILK
chemical_list: D009419:Nerve Tissue Proteins; D017346:Protein Serine-Threonine Kinases; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; D000099200:Scaffold Protein ILK; C497578:Nrep protein, rat; C508962:Tgfb1 protein, rat