PDF(Pubmed)
Abstract：
UNASSIGNED: Uveal melanoma is a poor prognosis cancer. Ergolide, a sesquiterpene lactone isolated from Inula Brittanica, exerts anti-cancer properties. The objective of this study was to 1) evaluate whether ergolide reduced metastatic uveal melanoma (MUM) cell survival/viability in vitro and in vivo; and 2) to understand the molecular mechanism of ergolide action.
UNASSIGNED: Ergolide bioactivity was screened via long-term proliferation assay in UM/MUM cells and in zebrafish MUM xenograft models. Mass spectrometry profiled proteins modulated by ergolide within whole cell or extracellular vesicle (EVs) lysates of the OMM2.5 MUM cell line. Protein expression was analyzed by immunoblots and correlation analyses to UM patient survival used The Cancer Genome Atlas (TCGA) data.
UNASSIGNED: Ergolide treatment resulted in significant, dose-dependent reductions (48.5 to 99.9%; p<0.0001) in OMM2.5 cell survival in vitro and of normalized primary zebrafish xenograft fluorescence (56%; p<0.0001) in vivo, compared to vehicle controls. Proteome-profiling of ergolide-treated OMM2.5 cells, identified 5023 proteins, with 52 and 55 proteins significantly altered at 4 and 24 hours, respectively ( p<0.05; fold-change >1.2). Immunoblotting of heme oxygenase 1 (HMOX1) and growth/differentiation factor 15 (GDF15) corroborated the proteomic data. Additional proteomics of EVs isolated from OMM2.5 cells treated with ergolide, detected 2931 proteins. There was a large overlap with EV proteins annotated within the Vesiclepedia compendium. Within the differentially expressed proteins, the proteasomal pathway was primarily altered. Interestingly, BRCA2 and CDKN1A Interacting Protein (BCCIP) and Chitinase Domain Containing 1 (CHID1), were the only proteins significantly differentially expressed by ergolide in both the OMM2.5 cellular and EV isolates and they displayed inverse differential expression in the cells versus the EVs.
UNASSIGNED: Ergolide is a novel, promising anti-proliferative agent for UM/MUM. Proteomic profiling of OMM2.5 cellular/EV lysates identified candidate pathways elucidating the action of ergolide and putative biomarkers of UM, that require further examination.
The most common form of adult eye cancer is uveal melanoma (UM). Once UM cancer cells spread to organs in the rest of the body, metastatic UM (MUM), there is a poor prognosis for patients with only one approved drug treatment. Hence, it is vital to better understand the cellular and extracellular proteins that regulate UM pathology in order to uncover biomarkers of disease and therapeutic targets. In this original study, we demonstrate a compound called ergolide is capable of severely reducing the metabolic activity and growth of UM cancer cells, grown as isolated monolayers. Ergolide was also able to reduce the growth of human MUM cells growing as tumors in transplanted zebrafish larvae. We identify that ergolide alters specific proteins found in the human UM cells. These proteins once analyzed in detail offer opportunities to understand how new treatment strategies can be developed for UM.
UNASSIGNED: Ergolide bioactivity was screened via long-term proliferation assay in UM/MUM cells and in zebrafish MUM xenograft models. Mass spectrometry profiled proteins modulated by ergolide within whole cell or extracellular vesicle (EVs) lysates of the OMM2.5 MUM cell line. Protein expression was analyzed by immunoblots and correlation analyses to UM patient survival used The Cancer Genome Atlas (TCGA) data.
UNASSIGNED: Ergolide treatment resulted in significant, dose-dependent reductions (48.5 to 99.9%; p<0.0001) in OMM2.5 cell survival in vitro and of normalized primary zebrafish xenograft fluorescence (56%; p<0.0001) in vivo, compared to vehicle controls. Proteome-profiling of ergolide-treated OMM2.5 cells, identified 5023 proteins, with 52 and 55 proteins significantly altered at 4 and 24 hours, respectively ( p<0.05; fold-change >1.2). Immunoblotting of heme oxygenase 1 (HMOX1) and growth/differentiation factor 15 (GDF15) corroborated the proteomic data. Additional proteomics of EVs isolated from OMM2.5 cells treated with ergolide, detected 2931 proteins. There was a large overlap with EV proteins annotated within the Vesiclepedia compendium. Within the differentially expressed proteins, the proteasomal pathway was primarily altered. Interestingly, BRCA2 and CDKN1A Interacting Protein (BCCIP) and Chitinase Domain Containing 1 (CHID1), were the only proteins significantly differentially expressed by ergolide in both the OMM2.5 cellular and EV isolates and they displayed inverse differential expression in the cells versus the EVs.
UNASSIGNED: Ergolide is a novel, promising anti-proliferative agent for UM/MUM. Proteomic profiling of OMM2.5 cellular/EV lysates identified candidate pathways elucidating the action of ergolide and putative biomarkers of UM, that require further examination.
The most common form of adult eye cancer is uveal melanoma (UM). Once UM cancer cells spread to organs in the rest of the body, metastatic UM (MUM), there is a poor prognosis for patients with only one approved drug treatment. Hence, it is vital to better understand the cellular and extracellular proteins that regulate UM pathology in order to uncover biomarkers of disease and therapeutic targets. In this original study, we demonstrate a compound called ergolide is capable of severely reducing the metabolic activity and growth of UM cancer cells, grown as isolated monolayers. Ergolide was also able to reduce the growth of human MUM cells growing as tumors in transplanted zebrafish larvae. We identify that ergolide alters specific proteins found in the human UM cells. These proteins once analyzed in detail offer opportunities to understand how new treatment strategies can be developed for UM.
摘要：
■葡萄膜黑色素瘤是一种预后不良的癌症。Ergolide,从Brittanica中分离出的倍半萜内酯，发挥抗癌特性。这项研究的目的是1）评估麦角利特是否在体外和体内降低转移性葡萄膜黑色素瘤（MUM）细胞的存活/活力；2）了解麦角利特作用的分子机制。
■通过长期增殖试验在UM/MUM细胞和斑马鱼MUM异种移植模型中筛选Ergolide生物活性。质谱分析在OMM2.5MUM细胞系的全细胞或细胞外囊泡（EV）裂解物中由麦高利特调节的蛋白质。通过免疫印迹分析蛋白质表达,并使用癌症基因组图谱(TCGA)数据对UM患者存活进行相关性分析。
■Ergolide治疗导致显著，剂量依赖性降低（48.5至99.9％；p<0.0001）在体外OM2.5细胞存活和在体内归一化的原代斑马鱼异种移植荧光（56％；p<0.0001），与车辆控制相比。麦高利特处理的OMM2.5细胞的蛋白质组分析，鉴定出5023种蛋白质，52和55蛋白质在4和24小时显著改变，分别(p<0.05；倍数变化>1.2)。血红素加氧酶1（HMOX1）和生长/分化因子15（GDF15）的免疫印迹证实了蛋白质组数据。从用麦高利德处理的OMM2.5细胞中分离出的EV的其他蛋白质组学，检测到2931种蛋白质。与Vesiclepedia汇编中注释的EV蛋白有很大的重叠。在差异表达的蛋白质中，蛋白酶体途径主要被改变。有趣的是,BRCA2和CDKN1A相互作用蛋白（BCCIP）和几丁质酶结构域1（CHID1），是在OMM2.5细胞和EV分离株中，ergolide显着差异表达的唯一蛋白质，并且它们在细胞中与EV中表现出相反的差异表达。
■Ergolide是一部小说，用于UM/MUM的有前途的抗增殖剂。OMM2.5细胞/EV裂解物的蛋白质组学分析确定了候选途径，阐明了ergolide的作用和UM的推定生物标志物，这需要进一步检查。
成人眼癌最常见的形式是葡萄膜黑色素瘤（UM）。一旦UM癌细胞扩散到身体其他部位的器官，转移性UM（MUM），只有一种批准的药物治疗的患者预后较差。因此，为了揭示疾病的生物标志物和治疗靶点,更好地了解调节UM病理的细胞和细胞外蛋白是至关重要的.在这项原始研究中，我们证明了一种名为ergolide的化合物能够严重降低UM癌细胞的代谢活性和生长，生长为孤立的单层。Ergolide还能够减少移植斑马鱼幼虫中作为肿瘤生长的人类MUM细胞的生长。我们确定麦戈力内酯会改变在人类UM细胞中发现的特定蛋白质。这些蛋白质一旦详细分析，就有机会了解如何为UM开发新的治疗策略。
■通过长期增殖试验在UM/MUM细胞和斑马鱼MUM异种移植模型中筛选Ergolide生物活性。质谱分析在OMM2.5MUM细胞系的全细胞或细胞外囊泡（EV）裂解物中由麦高利特调节的蛋白质。通过免疫印迹分析蛋白质表达,并使用癌症基因组图谱(TCGA)数据对UM患者存活进行相关性分析。
■Ergolide治疗导致显著，剂量依赖性降低（48.5至99.9％；p<0.0001）在体外OM2.5细胞存活和在体内归一化的原代斑马鱼异种移植荧光（56％；p<0.0001），与车辆控制相比。麦高利特处理的OMM2.5细胞的蛋白质组分析，鉴定出5023种蛋白质，52和55蛋白质在4和24小时显著改变，分别(p<0.05；倍数变化>1.2)。血红素加氧酶1（HMOX1）和生长/分化因子15（GDF15）的免疫印迹证实了蛋白质组数据。从用麦高利德处理的OMM2.5细胞中分离出的EV的其他蛋白质组学，检测到2931种蛋白质。与Vesiclepedia汇编中注释的EV蛋白有很大的重叠。在差异表达的蛋白质中，蛋白酶体途径主要被改变。有趣的是,BRCA2和CDKN1A相互作用蛋白（BCCIP）和几丁质酶结构域1（CHID1），是在OMM2.5细胞和EV分离株中，ergolide显着差异表达的唯一蛋白质，并且它们在细胞中与EV中表现出相反的差异表达。
■Ergolide是一部小说，用于UM/MUM的有前途的抗增殖剂。OMM2.5细胞/EV裂解物的蛋白质组学分析确定了候选途径，阐明了ergolide的作用和UM的推定生物标志物，这需要进一步检查。
成人眼癌最常见的形式是葡萄膜黑色素瘤（UM）。一旦UM癌细胞扩散到身体其他部位的器官，转移性UM（MUM），只有一种批准的药物治疗的患者预后较差。因此，为了揭示疾病的生物标志物和治疗靶点,更好地了解调节UM病理的细胞和细胞外蛋白是至关重要的.在这项原始研究中，我们证明了一种名为ergolide的化合物能够严重降低UM癌细胞的代谢活性和生长，生长为孤立的单层。Ergolide还能够减少移植斑马鱼幼虫中作为肿瘤生长的人类MUM细胞的生长。我们确定麦戈力内酯会改变在人类UM细胞中发现的特定蛋白质。这些蛋白质一旦详细分析，就有机会了解如何为UM开发新的治疗策略。
