Acta Anatomica Sinica

AAS ›› 2014, Vol. 45 ›› Issue (3): 393-397.doi: 10.3969/j.issn.0529-1356.2014.03.018

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Expression of interleukin -34/ colony stimulating factor -1R in the process of transforming growth factor -β1 inducing  A549 cells transition

ZHENG Jin-xu 1* ZHU Qin1 LIU Chao1 SONG Ping1 DUAN Li-rong 2   

  1. 1.Department of Respiratory Medicine, Affiliated Hospital of Jiangsu University, Jiangsu Zhenjiang 212001，China;2.College of Basic Medicine and Medical Technology，Jiangsu University， Jiangsu Zhenjiang 212013, China
  • Received:2014-02-18 Revised:2014-03-04 Online:2014-06-06 Published:2014-06-06
  • Contact: ZHENG Jin-xu E-mail:jxuzh135@163.com

Abstract:

Objective To investigate the expression of interleukin（IL)-34/ colony stimulating factor（CSF）-1R in the process of transforming growth factor（TGF）-β1 inducing epithelial-mesenchymal transition (EMT) of human alveolar epithelial cells A549 cells. Methods A549 cells were cultured in vitro. CCK 8 was used to test the influence of the proliferative rate of A549 cells which were stimulated by TGF-β1 at different concentrations and time points. A549 cells were stimulated by 5μg/L TGF-β1 at 0 hour, the 12th hour, the 24th hour, and the 48th hour. Western blotting was adopted to detect changes of the following proteins: α-smooth muscle actin(α-SMA), E-cadherin（E-Cad）, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1). Real-time PCR was adopted to detect changes of the following genes: IL-34 mRNA, CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA. Results TGF-β1 had no significant influence in the proliferation of A549 cells compared with the control group(P>0.05). TGF-β1（5μg/L）stimulated A549 cells at different time point (0 hour, 12, 24, 48 hours), compared with the control group. The epithelial phenotype E-Cad protein was gradually down-regulated(P<0.01), while the mesenchymal phenotype α-SMA protein was gradually up-regulated(P<0.01) and the protein of MMP-2 increased gradually (P<0.01). The protein of MMP-9 increased firstly and then was reduced(P<0.01),the peak was at the 24th hour. The protein of TIMP-1 was firstly transiently increased and then reduced(P<0.01)， the minimum was at the 48th hour. Compared with the control group, the gene of IL-34 mRNA increased gradually (P<0.01), and the genes of CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA increased firstly and then decreased(P<0.01)， which were peaked respectively at the 24th hour, the 24th hour, the 12th hour, respectively. Conclusion In the process of TGF-β1 inducing A549 cells transition，there is accompanied with the expression of IL-34/CSF-1R.

Key words: Transforming growth factor-β1, Interleukin-34/colony stimulating factor-1R, Pulmonary fibrosis, Epithlial-mesenchymal transition, CCK 8, Western blotting, Real-time PCR, A549 cell