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

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 A 549 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.