目的:探究核受体亚家族4A组成员1(nuclear receptor subfamily 4 group A member 1,Nr4a1)Nr4a1激动剂胞孢子酮B(cytosporone B,Csn-B)对小鼠噪声暴露后听力损失的治疗作用。方法:采用双氧水刺激HEI-OC1毛细胞系的方法构建氧化应激细胞...目的:探究核受体亚家族4A组成员1(nuclear receptor subfamily 4 group A member 1,Nr4a1)Nr4a1激动剂胞孢子酮B(cytosporone B,Csn-B)对小鼠噪声暴露后听力损失的治疗作用。方法:采用双氧水刺激HEI-OC1毛细胞系的方法构建氧化应激细胞模型;通过实时荧光定量PCR(quantitative real-time PCR,q PCR)检测细胞中Nr4a1的mRNA表达水平;分别通过细胞计数试剂盒(cell counting kit-8,CCK8)及流式细胞术的方法检测细胞活力和细胞凋亡水平以评估Csn-B预处理后经双氧水刺激的细胞状态。构建小鼠噪声性听力损失模型,运用qPCR和免疫荧光技术检测噪声暴露后Nr4a1在小鼠耳蜗中的表达;通过检测听性脑干反应(auditory brainstem response,ABR)评估噪声暴露后以及Csn-B连续治疗13 d后小鼠听力情况。结果:双氧水刺激后HEI-OC1毛细胞中Nr4a1表达上升,细胞活力显著下降,凋亡水平显著升高;Csn-B预处理HEI-OC1毛细胞经双氧水刺激,细胞活力显著高于对照组而凋亡水平则显著低于对照组。在体研究结果显示,噪声暴露后小鼠听力显著降低,Nr4a1在小鼠耳蜗中的表达水平显著升高。噪声暴露后经Csn-B治疗小鼠听力得到改善,主要表现为Click-ABR以及Tone Burst-ABR(4000、8000Hz处)阈值下降。结论:Nr4a1激动剂Csn-B增强内耳毛细胞对氧化应激损伤的抵御能力,部分改善噪声暴露后的小鼠听力。展开更多
Background:Pulmonary arterial hypertension(PAH)is a chronic and progressive disease that is strongly associated with dysregulation of glucose metabolism.Alterations in nuclear receptor subfamily 4 group A member 1(NR4...Background:Pulmonary arterial hypertension(PAH)is a chronic and progressive disease that is strongly associated with dysregulation of glucose metabolism.Alterations in nuclear receptor subfamily 4 group A member 1(NR4A1)activity alter the outcome of PAH.This study aimed to investigate the effects of NR4A1 on glycolysis in PAH and its underlying mechanisms.Methods:This study included twenty healthy volunteers and twenty-three PAH patients,and plasma samples were collected from the participants.To mimic the conditions of PAH in vitro,a hypoxia-induced model of pulmonary artery smooth muscle cell(PASMC)model was established.The proliferation of PASMCs was assessed using CCK8 assays.Results:Levels of NR4A1,hypoxia-inducible factor-1α(HIF-1α),and various glycolysis-related enzymes were measured.In addition,extracellular glucose and lactate production were assessed.The interaction between NR4A1 and HIF-1αwas evaluated by co-immunoprecipitation assays.Levels of NR4A1 and HIF-1αwas increased in PAH patients,and exposure to hypoxia resulted in increased levels of NR4A1 and HIF-1αin PASMCs.NR4A1 interacted with HIF-1α.NR4A1 overexpression enhanced hypoxia-induced expression of HIF-1α,GLUT1,PKM2,HK2,and CD36,decreased glucose levels,increased lactate levels and promoted hypoxic PASMC viability.Conversely,silencing NR4A1 decreased hypoxia-induced expression of HIF-1α,GLUT1,PKM2,HK2,and CD36,promoted glucose production,reduced lactate levels and inhibited hypoxic PASMC viability.Furthermore,overexpression of HIF-1αreversed the regulation of glycolysis caused by NR4A1 knockdown.Conclusion:NR4A1 enhances glycolysis in hypoxia-induced PASMCs by upregulating HIF-1α.Our findings indicate that the management of NR4A1 activity may be a promising strategy for PAH therapy.展开更多
SAM pointed domain containing E26 transformation-specific transcription factor(SPDEF)plays dual roles in the initiation and development of human malignancies.However,the biological role of SPDEF in head and neck squam...SAM pointed domain containing E26 transformation-specific transcription factor(SPDEF)plays dual roles in the initiation and development of human malignancies.However,the biological role of SPDEF in head and neck squamous cell carcinoma(HNSCC)remains unclear.In this study,the expression level of SPDEF and its correlation with the clinical parameters of patients with HNSCC were determined using TCGA-HNSC,GSE65858,and our own clinical cohorts.CCK8,colony formation,cell cycle analysis,and a xenograft tumor growth model were used to determine the molecular functions of SPDEF in HNSCC.ChIP-qPCR,dual luciferase reporter assay,and rescue experiments were conducted to explore the potential molecular mechanism of SPDEF in HNSCC.Compared with normal epithelial tissues,SPDEF was significantly downregulated in HNSCC tissues.Patients with HNSCC with low SPDEF mRNA levels exhibited poor clinical outcomes.Restoring SPDEF inhibited HNSCC cell viability and colony formation and induced G0/G1 cell cycle arrest,while silencing SPDEF promoted cell proliferation in vitro.The xenograft tumor growth model showed that tumors with SPDEF overexpression had slower growth rates,smaller volumes,and lower weights.SPDEF could directly bind to the promoter region of NR4A1 and promoted its transcription,inducing the suppression of AKT,MAPK,and NF-κB signaling pathways.Moreover,silencing NR4A1 blocked the suppressive effect of SPDEF in HNSCC cells.Here,we demonstrate that SPDEF acts as a tumor suppressor by transcriptionally activating NR4A1 in HNSCC.Our findings provide novel insights into the molecular mechanism of SPDEF in tumorigenesis and a novel potential therapeutic target for HNSCC.展开更多
基金supported by the National Natural Science Foundation of China(No.82000300).
文摘Background:Pulmonary arterial hypertension(PAH)is a chronic and progressive disease that is strongly associated with dysregulation of glucose metabolism.Alterations in nuclear receptor subfamily 4 group A member 1(NR4A1)activity alter the outcome of PAH.This study aimed to investigate the effects of NR4A1 on glycolysis in PAH and its underlying mechanisms.Methods:This study included twenty healthy volunteers and twenty-three PAH patients,and plasma samples were collected from the participants.To mimic the conditions of PAH in vitro,a hypoxia-induced model of pulmonary artery smooth muscle cell(PASMC)model was established.The proliferation of PASMCs was assessed using CCK8 assays.Results:Levels of NR4A1,hypoxia-inducible factor-1α(HIF-1α),and various glycolysis-related enzymes were measured.In addition,extracellular glucose and lactate production were assessed.The interaction between NR4A1 and HIF-1αwas evaluated by co-immunoprecipitation assays.Levels of NR4A1 and HIF-1αwas increased in PAH patients,and exposure to hypoxia resulted in increased levels of NR4A1 and HIF-1αin PASMCs.NR4A1 interacted with HIF-1α.NR4A1 overexpression enhanced hypoxia-induced expression of HIF-1α,GLUT1,PKM2,HK2,and CD36,decreased glucose levels,increased lactate levels and promoted hypoxic PASMC viability.Conversely,silencing NR4A1 decreased hypoxia-induced expression of HIF-1α,GLUT1,PKM2,HK2,and CD36,promoted glucose production,reduced lactate levels and inhibited hypoxic PASMC viability.Furthermore,overexpression of HIF-1αreversed the regulation of glycolysis caused by NR4A1 knockdown.Conclusion:NR4A1 enhances glycolysis in hypoxia-induced PASMCs by upregulating HIF-1α.Our findings indicate that the management of NR4A1 activity may be a promising strategy for PAH therapy.
基金This work was supported by grants from the National Natural Science Foundation of China(81870769,81702700)the Guangdong Financial Fund for High-Caliber Hospital Construction(174-2018-XMZC-0001-03-0125/D-05,174-2018-XMZC-0001-03-0125/C-08)+1 种基金the Natural Science Foundation of Guangdong Province(2019A1515010679)the Fundamental Research Funds for the Central Universities(19ykpy83).
文摘SAM pointed domain containing E26 transformation-specific transcription factor(SPDEF)plays dual roles in the initiation and development of human malignancies.However,the biological role of SPDEF in head and neck squamous cell carcinoma(HNSCC)remains unclear.In this study,the expression level of SPDEF and its correlation with the clinical parameters of patients with HNSCC were determined using TCGA-HNSC,GSE65858,and our own clinical cohorts.CCK8,colony formation,cell cycle analysis,and a xenograft tumor growth model were used to determine the molecular functions of SPDEF in HNSCC.ChIP-qPCR,dual luciferase reporter assay,and rescue experiments were conducted to explore the potential molecular mechanism of SPDEF in HNSCC.Compared with normal epithelial tissues,SPDEF was significantly downregulated in HNSCC tissues.Patients with HNSCC with low SPDEF mRNA levels exhibited poor clinical outcomes.Restoring SPDEF inhibited HNSCC cell viability and colony formation and induced G0/G1 cell cycle arrest,while silencing SPDEF promoted cell proliferation in vitro.The xenograft tumor growth model showed that tumors with SPDEF overexpression had slower growth rates,smaller volumes,and lower weights.SPDEF could directly bind to the promoter region of NR4A1 and promoted its transcription,inducing the suppression of AKT,MAPK,and NF-κB signaling pathways.Moreover,silencing NR4A1 blocked the suppressive effect of SPDEF in HNSCC cells.Here,we demonstrate that SPDEF acts as a tumor suppressor by transcriptionally activating NR4A1 in HNSCC.Our findings provide novel insights into the molecular mechanism of SPDEF in tumorigenesis and a novel potential therapeutic target for HNSCC.
