干扰TAK1通过下调TAB2/3来抑制肺成纤维细胞的增殖、侵袭、迁移和纤维化  

TAK1 interference inhibiting TGF-β1 induced proliferation,invasion,migration and fibrosis in lung fibroblasts via regulation of TAB2/3

作　　者：章艳菊[1] 周小娣[1] 夏云飞[2] ZHANG Yanju;ZHOU Xiaodi;XIA Yunfei(Infection Management Office;Department of Rheumatology and Immunology,the Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001,China)

机构地区：[1]南通大学附属医院感染管理办公室,江苏南通226001 [2]南通大学附属医院风湿免疫科,江苏南通226001

出　　处：《临床肺科杂志》2023年第10期1523-1530,共8页Journal of Clinical Pulmonary Medicine

摘　　要：目的探讨转化生长因子β激活激酶1(TAK1)对转化生长因子β1(TGF-β1)诱导的人胚肺成纤维细胞(HFL-1)的增殖、侵袭、迁移和纤维化的影响及其与TAK1结合蛋白(TAB)2/3的关系。方法将细胞分组分为Control组(未处理HFL-1)、TGF-β1组(10ng/mL TGF-β1诱导HFL-1)、TGF-β1+si-NC组(转染si-NC+TGF-β1诱导)、TGF-β1+si-TAK1组(转染si-TAK1+TGF-β1诱导)、TGF-β1+si-TAK1+pcDNA3.1组(转染si-TAK1+pcDNA3.1+TGF-β1诱导)、TGF-β1+si-TAK1+pcDNA3.1-TAB2组(转染si-TAK1+pcDNA3.1-TAB2+TGF-β1诱导)、TGF-β1+si-TAK1+pcDNA3.1-TAB3组(转染si-TAK1+pcDNA3.1-TAB3+TGF-β1诱导)。采用Western Blot和RT-qPCR检测TAK1和TAB2/3的表达。CCK-8检测细胞增殖。Transwell和细胞划痕实验检测细胞的侵袭和迁移能力。Western Blot检测迁移和纤维化相关蛋白的表达。免疫共沉淀验证TAK1和TAB2/3之间的关联。结果与Control组比,TGF-β1诱导后TAK1及TAB2/3表达升高(P均<0.05)。TGF-β1+si-TAK1组与TGF-β1+si-NC组相比其增殖、侵袭、迁移能力明显下降,迁移及纤维化相关蛋白表达明显下降(P均<0.05)。TGF-β1+si-TAK1+pcDNA3.1-TAB2/3组与TGF-β1+si-TAK1+pcDNA3.1组比增殖、侵袭、迁移能力上升,迁移及纤维化相关蛋白表达上升(P<0.05)。结论干扰TAK1通过下调TAB2/3来抑制TGF-β1诱导的肺成纤维细胞的增殖、侵袭、迁移和纤维化。Objective To investigate the effect of transforming growth factor-βactivated kinase 1(TAK1)on the proliferation,invasion,migration and fibrosis of Human fetal lung fibroblast 1(HFL-1)induced by transforming growth factor-β1(TGF-β1)and its relationship with TAK1 binding protein(TAB)2/3.Methods The cells were divided into the control group(no HFL-1),the TGF-β1 group(HFL-1 induction 10ng/mL TGF-β1),the TGF-β1+si-NC group(transfection si-NC and TGF-β1 induction),the TGF-β1+si-TAK1 group(transfection si-TAK1 and TGF-β1 induction),the TGF-β1+si-TAK1+pcDNA3.1 group(transfection si-TAK1+pcDNA3.1 and TGF-β1 induction),the TGF-β1+si-TAK1+pcDNA3.1-TAB2 group(transfection si-TAK1+pcDNA3.1-TAB2 and TGF-β1 induction),and the TGF-β1+si-TAK1+pcDNA3.1-TAB3 group(transfection si-TAK1+pcDNA3.1-TAB3 and TGF-β1 induction).The expression of TAK1 and TAB2/3 were detected by western blot analysis and reverse transcription-quantitative PCR(RT-qPCR).Cell proliferation was detected by Cell Counting Kit-8 assay.Wound healing and Transwell assays were used to detect cell migration and invasion,and western blot was used to detect the expression of migration and fibrosis-related proteins.The association between TAK1 and TAB2/3 in cells was verified by co-immunoprecipitation.Results The cell proliferation,invasion and migration of the TGF-β1+si-TAK1 group were significantly lower than those of the TGF-β1+si-NC group(P<0.05).The expression of migration and fibrosis-related proteins were decreased(P<0.05).The cell proliferation,invasion,and migration of the TGF-β1+si-TAK1+pcDNA3.1-TAB2/3 group were higher than the TGF-β1+si-TAK1+pcDNA3.1 group(P<0.05).The expression of migration and fibrosis-related proteins were increased(P<0.05).Conclusion TAK1 interference inhibits the TGF-β1-induced proliferation,invasion,migration and fibrosis of lung fibroblasts via regulation of TAB2/3.

关 键 词：转化生长因子β激活激酶1 TAK1结合蛋白2/3 肺纤维化 侵袭 迁移 

分 类 号：R563[医药卫生—呼吸系统]