Effect of Snail-mediated lung epithelial-mesenchymal transition in activation of myofibroblasts

doi:10.3969/j.issn.1006-7795.2020.01.018

Abstract

Abstract: Objective To explore the effect of nuclear transcription factor Snail-mediated lung epithelial-mesenchymal transition (EMT) in activation of myofibroblasts. Methods MLE-12 cells (murine epithelial cell line) were stimulated by transforming growth factor-β1 (TGF-β1) to build EMT model, the mRNA expression of epithelial marker CDH1, mesenchymal marker vimentin and nuclear transcription factor Snail were detected by RT-qPCR. MLE-12 cells were co-cultured with mouse embryo fibroblast (NIH-3T3) cells. The mRNA expression of α-smooth muscle actin (α-SMA), collagen Ⅰ α1(COL1A1), collagen Ⅲ α1(COL3A1) of NIH-3T3 were detected by RT-qPCR. MLE-12 cells were transfected with Snail-shRNA lentivirus. The expression of Snail mRNA and protein of MLE-12 cells were detected by RT-qPCR and Western blotting. The Snail-shRNA-transfected-MLE-12 cells were co-cultured with NIH-3T3 cells. The mRNA expression of α-SMA, COL1A1, COL3A1 of NIH-3T3 were detected by RT-qPCR. Multiple groups were analyzed by one-way ANOVA and LSD test to determine significant differences between groups at P<0.05. Results The results of RT-qPCR showed that after stimulated by TGF-β1 for 48 hours, compared with control, the mRNA expression of CDH1 of MLE-12 in TGF-β1 group was down-regulated while α-SMA and Snail were up-regulated (P<0.05). In the co-culture model, the results of RT-qPCR showed that compared to TGF-β1 and MLE-12 group, MLE-12 stimulated by TGF-β1 cause up-regulation of α-SMA, COL1A1, COL3A1 mRNA expression of NIH-3T3 (P<0.05). After transfected with Snail-shRNA lentivirus, mRNA and protein expression of MLE-12 were down-regulated compared with control virus group (P<0.05). In the co-culture model of lentivirus-transfected-MLE-12 and NIH-3T3, results of RT-qPCR showed that α-SMA, COL1A1, COL3A1 mRNA up-regulation of NIH-3T3 in TGF-β1+Snail-shRNA group were lower than TGF-β1+control-shRNA group (P<0.05). Conclusion Snail-mediated EMT of MLE-12 can cause activation of myofibroblasts, knockdown of Snail in MLE-12 can inhibit fibroblasts activation into myofibroblasts, suggested Snail-mediated lung epithelial-mesenchymal transition plays an important role in myofibroblasts activation.

Key words: Snail, epithelial-mesenchymal transition, fibroblast, myofibroblast

CLC Number:

Li Siling, Zhu Zhonghui, Li Qiuyue, Xu Chunjie, Zhao Jing, Wang Yan, Tian Lin. Effect of Snail-mediated lung epithelial-mesenchymal transition in activation of myofibroblasts[J]. Journal of Capital Medical University, 2020, 41(1): 92-98.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2020.01.018

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2020/V41/I1/92

References

[1] Para R, Romero F, George G, et al. Metabolic reprogramming as a driver of fibroblast activation in pulmonary fibrosis[J]. Am J Med Sci, 2019,357(5):394-398.
[2] Chapman H A. Epithelial-mesenchymal interactions in pulmonary fibrosis[J]. Annu Rev Physiol, 2011,73:413-435.
[3] Kim K K, Kugler M C, Wolters P J, et al. Alveolar epithelial cell mesenchymal transition develops in vivo during pulmonary fibrosis and is regulated by the extracellular matrix[J]. Proc Natl Acad Sci U S A, 2006,103(35):13180-13185.
[4] Sugimoto H, LeBleu V S, Bosukonda D, et al. Activin-like kinase 3 is important for kidney regeneration and reversal of fibrosis[J]. Nat Med, 2012,18(3):396-404.
[5] Wang Y, Liang D, Zhu Z, et al. Bone morphogenetic protein-7 prevented epithelial-mesenchymal transition in RLE-6TN cells[J]. Toxicol Res (Camb), 2016,5(3):931-937.
[6] Wang Y, Yang G, Zhu Z, et al. Effect of bone morphogenic protein-7 on the expression of epithelial-mesenchymal transition markers in silicosis model[J]. Exp Mol Pathol, 2015,98(3):393-402.
[7] Tanjore H, Xu X C, Polosukhin V V, et al. Contribution of epithelial-derived fibroblasts to bleomycin-induced lung fibrosis[J]. Am J Respir Crit Care Med, 2009,180(7):657-665.
[8] Richeldi L, Collard H R, Jones M G. Idiopathic pulmonary fibrosis[J]. Lancet, 2017,389(10082):1941-1952.
[9] Raghu G, Collard H R, Egan J J, et al. An official ATS/ERS/JRS/ALAT statement:idiopathic pulmonary fibrosis:evidence-based guidelines for diagnosis and management[J]. Am J Respir Crit Care Med, 2011,183(6):788-824.
[10] Duck A, Pigram L, Errhalt P, et al. IPF care:a support program for patients with idiopathic pulmonary fibrosis treated with pirfenidone in Europe[J]. Adv Ther, 2015,32(2):87-107.
[11] Thannickal V J. Mechanisms of pulmonary fibrosis:role of activated myofibroblasts and NADPH oxidase[J]. Fibrogenesis Tissue Repair, 2012,5(Suppl 1):S23.
[12] Rennard S I, Bitterman P B, Crystal R G. Response of the lower respiratory tract to injury. Mechanisms of repair of the parenchymal cells of the alveolar wall[J]. Chest, 1983,84(6):735-739.
[13] Fletcher S, Jones M G, Spinks K, et al. The safety of new drug treatments for idiopathic pulmonary fibrosis[J]. Expert Opin Drug Saf, 2016,15(11):1483-1489.
[14] Iwano M, Plieth D, Danoff T M, et al. Evidence that fibroblasts derive from epithelium during tissue fibrosis[J]. J Clin Invest, 2002,110(3):341-350.
[15] Zeisberg M, Yang C, Martino M, et al. Fibroblasts derive from hepatocytes in liver fibrosis via epithelial to mesenchymal transition[J]. J Biol Chem, 2007,282(32):23337-23347.
[16] Foroutan M, Cursons J, Hediyeh-Zadeh S, et al. A transcriptional program for detecting TGF beta-induced EMT in cancer[J]. Mol Cancer Res, 2017,15(5):619-631.
[17] Leung C C, Yu I T, Chen W. Silicosis[J]. Lancet, 2012,379(9830):2008-2018.
[18] Lovisa S, LeBleu V S, Tampe B, et al. Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis[J]. Nat Med, 2015,21(9):998-1009.
[19] Sun J, Li Q, Lian X, et al. MicroRNA-29b mediates lung mesenchymal-epithelial transition and prevents lung fibrosis in the silicosis model[J]. Mol Ther Nucleic Acids, 2019,14:20-31.
[20] Feng H, Lu J J, Wang Y, et al. Osthole inhibited TGF beta-induced epithelial-mesenchymal transition (EMT) by suppressing NF-kappaB mediated Snail activation in lung cancer A549 cells[J]. Cell Adh Migr, 2017,11(5-6):464-475.
[21] Hu Y B, Li F F, Deng Z H, et al. Transcriptional factor snail mediates epithelial-mesenchymal transition in human bronchial epithelial cells induced by silica[J]. Biomed Environ Sci, 2015,28(7):544-548.