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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* ZHU Qin LIU Chao SONG Ping DUAN Li-rong
Acta Anatomica Sinica ›› 2014, Vol. 45 ›› Issue (3) : 393-397.
Expression of interleukin -34/ colony stimulating factor -1R in the process of transforming growth factor -β1 inducing A549 cells transition
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.
Transforming growth factor-β1 / Interleukin-34/colony stimulating factor-1R / Pulmonary fibrosis / Epithlial-mesenchymal transition / CCK 8 / Western blotting / Real-time PCR / A549 cell
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