Tranilast attenuates TGF-beta1-induced epithelial-mesenchymal transition in the NRK-52E cells

We previously reported that tranilast can halt the pathogenesis of chronic cyclosporine nephrotoxicity in rats via the transforming growth factor-beta [TGF-beta] /Smad pathway, an important signaling system involved in epithelialmesenchymal transition [EMT], but the exact underlying cellular mechanisms are not yet clear. Thus, by selecting [0]TGF-beta1-induced normal rat kidney proximal tubular epithelial cells [NRK-52E] as a model, we demonstrated potential modifying effect of tranilast on EMT-induced by TGF-beta1 in vitro. NRK-52E cells were incubated with the blank vehicle [Dulbecco's modified Eagle's medium and F-12 [DMEM/F12] added with 10% fetal bovine serum [FBS]], 10 ng/ml TGF-beta1 alone or together with 100, 200 or 400microM tranilast for 48 h after incubation in medium containing 1% FBS for 24 h. Cell morphological changes were observed to confirm occurrence of EMT. Protein expressions of two typical markers of EMT, E-cadherin and alpha-smooth muscle actin [alpha-SMA], were assessed by western blotting and flow cytometry, respectively. Our results showed that TGF-beta1 induced spindle-like morphological transition, the loss of Ecadherin protein and upregulation of expression of alpha-SMA. However, the TGF-beta1-produced changes in cellular morphology, E-cadherin and alpha-SMA were inversed by tranlilast in concentration-dependent manner. Our findings indicate that tranilast can directly inhibit EMT. Thus, it may be implied that regulation of EMT be the target to prevent renal tubulointerstitial fibrosis.

Research on the effects of PIAS3 expression on the invasion of glioma TJ905 cells / 中华外科杂志

<p><b>OBJECTIVES</b>To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells.</p><p><b>METHODS</b>PIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or downregulation of PIAS3 expression levels in TJ905 cells. After that, the invasive effects of TJ905 cells were measured by Transwell assay, and the expression of PIAS3, tissue inhibitor of metalloproteinases (TIMP)3, matrix metalloprotease (MMP)-2, and MMP-9 were identified by Western blot.</p><p><b>RESULTS</b>In vitro transfection efficiency of plasmids and oligonucleotides were separately 85.3% ± 3.1% and 95.1% ± 2.9%. PIAS3 overexpression plasmid transfection in vitro could effectively improve the expression of PIAS3 protein in TJ905 cells and inhibit the invasion of TJ905 cells (P < 0.05), and cell penetration ratio reduced from 87.9% ± 9.3% to 37.3% ± 7.9% compared with control group, while it upregulated TIMP3 and downregulated MMP-2, MMP-9 protein expression (P < 0.05); PIAS3 siRNA transfection could inhibit the PIAS3 protein expression of TJ905 cells and promote the invasion of TJ905 cells (P < 0.05), and cell penetration ratio increased from 83.9% ± 7.1% to 93.2% ± 3.1% compared with control group, while it downregulated TIMP3 and upregulated MMP-2, MMP-9 protein expression (P < 0.05).</p><p><b>CONCLUSION</b>PIAS3 expression is closely related to the invasion properties of glioma TJ905 cells.</p>