CAJ | 학술논문

目的：探讨白细胞介素-34/集落刺激因子-1受体（IL-34/CSF-1R）在转化生长因子-β1（TGF-β1）诱导A549细胞发生上皮-间质转化（ EMT）中的表达。方法体外培养A549细胞；CCK 8法检测不同浓度TGF-β1在不同时间点对A549细胞增殖的影响；选用5μg/L TGF-β1刺激A549细胞（0、12、24、48h），提取细胞蛋白和RNA， Western blotting法检测α-平滑肌肌动蛋白（α-SMA）、E-钙黏连蛋白（E-Cad）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（ MMP-9）、基质金属蛋白酶抑制剂-1（ TIMP-1）蛋白的变化；Real-time PCR 法检测 IL-34、CSF-1R、MMP-2、MMP-9基因表达的变化。结果 TGF-β1对A549细胞的增殖无明显影响（ P＞0.05）。 TGF-β1刺激A549细胞后，间质细胞标志性蛋白α-SMA表达升高，上皮细胞标志性蛋白E-Cad表达下降，纤维化相关的MMP-2蛋白表达升高，MMP-9蛋白表达先升高后下降，TIMP-1蛋白早期出现短暂性增高后，呈持续降低趋势（ P＜0.01）。 IL-34 mRNA表达逐渐升高，CSF-1R mRNA、MMP-2 mRNA、MMP-9 mRNA表达先升高后下降（P＜0.01）。结论 TGF-β1诱导A549细胞的EMT过程中，存在IL-34/CSF-1R的动态表达。
목적：탐토백세포개소-34/집락자격인자-1수체（IL-34/CSF-1R）재전화생장인자-β1（TGF-β1）유도A549세포발생상피-간질전화（ EMT）중적표체。방법체외배양A549세포；CCK 8법검측불동농도TGF-β1재불동시간점대A549세포증식적영향；선용5μg/L TGF-β1자격A549세포（0、12、24、48h），제취세포단백화RNA， Western blotting법검측α-평활기기동단백（α-SMA）、E-개점련단백（E-Cad）、기질금속단백매-2（MMP-2）、기질금속단백매-9（ MMP-9）、기질금속단백매억제제-1（ TIMP-1）단백적변화；Real-time PCR 법검측 IL-34、CSF-1R、MMP-2、MMP-9기인표체적변화。결과 TGF-β1대A549세포적증식무명현영향（ P＞0.05）。 TGF-β1자격A549세포후，간질세포표지성단백α-SMA표체승고，상피세포표지성단백E-Cad표체하강，섬유화상관적MMP-2단백표체승고，MMP-9단백표체선승고후하강，TIMP-1단백조기출현단잠성증고후，정지속강저추세（ P＜0.01）。 IL-34 mRNA표체축점승고，CSF-1R mRNA、MMP-2 mRNA、MMP-9 mRNA표체선승고후하강（P＜0.01）。결론 TGF-β1유도A549세포적EMT과정중，존재IL-34/CSF-1R적동태표체。
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.