目的:探讨细胞命运决定因子DACH1(dachshund family transcription factor 1)对人子宫内膜癌HEC-1A细胞体外增殖能力和细胞周期的影响。方法 :将DACH1过表达的重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞,在激光共聚焦显微镜下观...目的:探讨细胞命运决定因子DACH1(dachshund family transcription factor 1)对人子宫内膜癌HEC-1A细胞体外增殖能力和细胞周期的影响。方法 :将DACH1过表达的重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞,在激光共聚焦显微镜下观察DACH1在HEC-1A细胞中的定位;分别采用实时荧光定量PCR法和蛋白质印迹法检测HEC-1A细胞中DACH1 m RNA和蛋白的表达,MTT法检测HEC-1A细胞的体外的增殖能力,FCM法检测细胞周期的分布。结果 :重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞后,LV-DACH1-GFP主要分布在细胞核中。LV-DACH1-GFP感染组HEC-1A细胞中,DACH1 m RNA和蛋白的表达水平均明显高于对照组(HEC-1A细胞感染慢病毒LV-GFP)和空白组(HEC-1A细胞未进行任何感染)(P值均<0.01),LV-DACH1-GFP感染组HEC-1A细胞的增殖受到明显抑制(P<0.05),细胞周期阻滞于G0/G1期(P<0.01)。结论 :D ACH1基因可抑制人子宫内膜癌HEC-1A细胞的增殖,这一作用可能与DACH1基因阻滞细胞周期有关。展开更多
Idiopathic pulmonary fibrosis(IPF)is a progressive disease with unknown etiology and limited therapeutic options.Activation of fibroblasts is a prominent feature of pulmonary fibrosis.Here we report that lncRNA DACH1(...Idiopathic pulmonary fibrosis(IPF)is a progressive disease with unknown etiology and limited therapeutic options.Activation of fibroblasts is a prominent feature of pulmonary fibrosis.Here we report that lncRNA DACH1(dachshund homolog 1)is downregulated in the lungs of IPF patients and in an experimental mouse model of lung fibrosis.LncDACH1 knockout mice develop spontaneous pulmonary fibrosis,whereas overexpression of LncDACH1 attenuated TGF-β1-induced aberrant activation,collagen deposition and differentiation of mouse lung fibroblasts.Similarly,forced expression of LncDACH1 not only prevented bleomycin(BLM)-induced lung fibrosis,but also reversed established lung fibrosis in a BLM model.Mechanistically,LncDACH1 binding to the serine/arginine-rich splicing factor 1(SRSF1)protein decreases its activity and inhibits the accumulation of Ctnnb1.Enhanced expression of SRSF1 blocked the anti-fibrotic effect of LncDACH1 in lung fibroblasts.Furthermore,loss of LncDACH1 promoted proliferation,differentiation,and extracellular matrix(ECM)deposition in mouse lung fibroblasts,whereas such effects were abolished by silencing of Ctnnb1.In addition,a conserved fragment of LncDACH1 alleviated hyperproliferation,ECM deposition and differentiation of MRC-5 cells driven by TGF-β1.Collectively,LncDACH1 inhibits lung fibrosis by interacting with SRSF1 to suppress CTNNB1 accumulation,suggesting that LncDACH1 might be a potential therapeutic target for pulmonary fibrosis.展开更多
文摘目的:探讨细胞命运决定因子DACH1(dachshund family transcription factor 1)对人子宫内膜癌HEC-1A细胞体外增殖能力和细胞周期的影响。方法 :将DACH1过表达的重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞,在激光共聚焦显微镜下观察DACH1在HEC-1A细胞中的定位;分别采用实时荧光定量PCR法和蛋白质印迹法检测HEC-1A细胞中DACH1 m RNA和蛋白的表达,MTT法检测HEC-1A细胞的体外的增殖能力,FCM法检测细胞周期的分布。结果 :重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞后,LV-DACH1-GFP主要分布在细胞核中。LV-DACH1-GFP感染组HEC-1A细胞中,DACH1 m RNA和蛋白的表达水平均明显高于对照组(HEC-1A细胞感染慢病毒LV-GFP)和空白组(HEC-1A细胞未进行任何感染)(P值均<0.01),LV-DACH1-GFP感染组HEC-1A细胞的增殖受到明显抑制(P<0.05),细胞周期阻滞于G0/G1期(P<0.01)。结论 :D ACH1基因可抑制人子宫内膜癌HEC-1A细胞的增殖,这一作用可能与DACH1基因阻滞细胞周期有关。
基金supported by the National Natural Science Foundation of China (32171127 and 91949109)the HMU Marshal Initiative Funding (HMUMIF-21023, China)+3 种基金the Major Scientific Fund Project of Heilongjiang Province (ZD2019H001, China)the CAMS Innovation Fund for Medical Sciences (CIFMS, 2019-I2M5-078, China)the Guangdong Province Basic and Applied Basic Research Fund (2021A1515111049, China)Postgraduate Research and Practice Innovation Program of HMU (YJSCX202015HYD, China)
文摘Idiopathic pulmonary fibrosis(IPF)is a progressive disease with unknown etiology and limited therapeutic options.Activation of fibroblasts is a prominent feature of pulmonary fibrosis.Here we report that lncRNA DACH1(dachshund homolog 1)is downregulated in the lungs of IPF patients and in an experimental mouse model of lung fibrosis.LncDACH1 knockout mice develop spontaneous pulmonary fibrosis,whereas overexpression of LncDACH1 attenuated TGF-β1-induced aberrant activation,collagen deposition and differentiation of mouse lung fibroblasts.Similarly,forced expression of LncDACH1 not only prevented bleomycin(BLM)-induced lung fibrosis,but also reversed established lung fibrosis in a BLM model.Mechanistically,LncDACH1 binding to the serine/arginine-rich splicing factor 1(SRSF1)protein decreases its activity and inhibits the accumulation of Ctnnb1.Enhanced expression of SRSF1 blocked the anti-fibrotic effect of LncDACH1 in lung fibroblasts.Furthermore,loss of LncDACH1 promoted proliferation,differentiation,and extracellular matrix(ECM)deposition in mouse lung fibroblasts,whereas such effects were abolished by silencing of Ctnnb1.In addition,a conserved fragment of LncDACH1 alleviated hyperproliferation,ECM deposition and differentiation of MRC-5 cells driven by TGF-β1.Collectively,LncDACH1 inhibits lung fibrosis by interacting with SRSF1 to suppress CTNNB1 accumulation,suggesting that LncDACH1 might be a potential therapeutic target for pulmonary fibrosis.
