Roles of p38 MAPK and JNK in TGF-β1-induced human alveolar epithelial to mesenchymal transition.

Idiopathic pulmonary fibrosis (IPF) is associated with significant morbidity and mortality despite aggressive therapy. The aim of the present study is to investigate the roles of p38 MAPK and JNK in TGF-β1-induced human alveolar epithelial to mesenchymal transition (EMT), which could be a possible mechanism of IPF. A549 cells were treated with TGF-β1 (3 ng/mL) for 48 h to induce EMT. The expression of mesenchymal phenotypic markers including desmin, α-smooth muscle actin (α-SMA) and vimentin, and expression of epithelial phenotypic markers including E-cadherin, zonula occludens-1 (ZO-1) and aquaporin-5 (AQP5) were detected by Western blot. The roles of p38 MAPK and JNK in TGF-β1-mediated EMT were investigated using gene silencing and inhibitor SB-203580 and SP-600125. The data showed that TGF-β1 induced A549 cells with an alveolar epithelial type II cell phenotype to undergo EMT. The process of EMT was accompanied by morphological alteration and expression of the myofibroblast marker desmin, α-SMA and vimentin, concomitant with a downregulation of the epithelial cell marker E-cadherin, ZO-1 and AQP5. TGF-β1-induced EMT occurred through phosphorylation of p38 MAPK and JNK and was inhibited by inhibitor SB-203580 and SP-600125 and gene silencing. TGF-β1 induces A549 alveolar epithelial cells (AECs) to undergo EMT partially via p38 MAPK and JNK activation and supports the concept of EMT in lung epithelial cells.

Chen HH ，Zhou XL ，Shi YL ，Yang J ... -  《-》

Transglutaminase-2 induces N-cadherin expression in TGF-β1-induced epithelial mesenchymal transition via c-Jun-N-terminal kinase activation by protein phosphatase 2A down-regulation.

Epithelial-mesenchymal-transition (EMT) is a key event for tumour cells to initiate metastasis leading to switching of E-cadherin to N-cadherin. Transglutaminase-2 (Tgase-2) expression is increased in TGF-β1-induced EMT in A549 lung cancer cells or other lung cancer cells. The role and underlying mechanism of Tgase-2 in N-cadherin switching of TGF-β1-induced EMT are not known. The involvement and mechanisms of Tgase-2 were investigated in A549 cells using chemical inhibitors, gene silencing and over-expression. TGF-β1-induced EMT was suppressed by cystamine or gene silencing of Tgase-2. Suppression of Tgase-2 or the c-Jun-N-terminal kinase (JNK) inhibitor, SP600125, significantly reduced and over-expression of Tgase-2 increased the expression of N-cadherin. The relationship between Tgase-2 and JNK in the TGF-β1-induced EMT of A549 cells was examined using Tgase-2 over-expressed A549 cells (A549(TG2)) and Tgase-2 silenced A549 cells (A549(shTG2)). JNK activation was significantly increased in A549(TG2) cells and decreased in A549(shTG2) cells. In contrast, PP2A expression was decreased in A549(TG2) and A549 cells and increased in A549(shTG2) cells. The involvement of Tgase-2 in N-cadherin expression was also confirmed in an in vivo lung cancer orthotopic model by injection of A549(WT) and A549(shTG2) cells into SCID mice. Tgase-2 expressing A549(WT) cells-injected mice group showed increased expressions of N-cadherin and JNK activation, but decreased expression of PP2A in lung cancer tissue comparing with the A549(shTG2) cells-injected group. These results suggested that Tgase-2 induces N-cadherin expression of TGF-β1-induced EMT via JNK activation by PP2A down-regulation, and Tgase-2/PP2A/JNK might be a novel axis that affects N-cadherin switching in the EMT of A549 lung cancer cells.

Park MK ，You HJ ，Lee HJ ，Kang JH ，Oh SH ，Kim SY ，Lee CH ... -  《-》

TGF-beta1 induces human alveolar epithelial to mesenchymal cell transition (EMT).

Kasai H ，Allen JT ，Mason RM ，Kamimura T ，Zhang Z ... -  《RESPIRATORY RESEARCH》

Simvastatin attenuates TGF-β1-induced epithelial-mesenchymal transition in human alveolar epithelial cells.

Transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of alveolar epithelial cells (AEC) may contribute to idiopathic pulmonary fibrosis (IPF). TGF-β1-induced EMT in A549 cells (a human AEC cell line) resulted in the adoption of mesenchymal responses that were predominantly mediated via the TGF-β1-Smad2/3 signaling pathway. Simvastatin (Sim), a 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase inhibitor, has been previously reported to inhibit EMT in human proximal tubular epithelial cells and porcine lens epithelial cells and to suppress Smad2/3 phosphorylation in animal models. However, whether Sim can attenuate TGF-β1-induced EMT in A549 cells and its underlying mechanisms remains unknown. Cells were incubated with TGF-β1 in the presence or absence of Sim. The epithelial marker E-cadherin (E-Cad) and the mesenchymal markers, α-smooth muscle actin (α-SMA), vimentin (Vi) and fibronectin (FN), were detected using western blotting analyses and immunofluorescence. Phosphorylated Smad2 and Smad3 levels and connective tissue growth factor (CTGF) were analyzed using western blotting. In addition, a cell migration assay was performed. Moreover, the levels of matrix metalloproteinase (MMP)-2 and -9 in the culture medium were examined using ELISA. Sim significantly attenuated the TGF-β1-induced decrease in E-Cad levels and elevated the levels of α-SMA, Vi and FN via the suppression of Smad2 and Smad3 phosphorylation. Furthermore, Sim inhibited the mesenchymal-like responses in A549 cells, including cell migration, CTGF expression and secretion of MMP-2 and -9. However, Sim failed to reverse the cell morphologial changes induced by TGF-β1 in A549 cells. Sim attenuated TGF-β1-induced EMT in A549 cells and might be a promising therapeutic agent for treating IPF.

Yang T ，Chen M ，Sun T 《-》

Role of reactive oxygen species in TGF-beta1-induced mitogen-activated protein kinase activation and epithelial-mesenchymal transition in renal tubular epithelial cells.

Rhyu DY ，Yang Y ，Ha H ，Lee GT ，Song JS ，Uh ST ，Lee HB ... -  《JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY》