The balance between inflammatory T helper type 17(Th17)and immunosuppressive regulatory T(Treg)cells is critical for maintaining immune homeostasis in the human body and is tightly regulated under healthy conditions.A...The balance between inflammatory T helper type 17(Th17)and immunosuppressive regulatory T(Treg)cells is critical for maintaining immune homeostasis in the human body and is tightly regulated under healthy conditions.An increasing number of studies have reported that deubiquitinases(DUBs)play a vital role in regulating Th17-and Treg-cell differentiation.However,the biological functions of only a small fraction of DUBs in Th17-and Treg-cell differentiation are well defined.In this study,we identified ubiquitin-specific peptidase 1(USP1)as a vital regulator of CD4+T-cell differentiation.USP1 promoted Th17-cell differentiation but attenuated Treg-cell differentiation,thereby promoting the development of inflammatory diseases.Mechanistically,USP1 in CD4+T cells enhanced the activity of RORγt but promoted the proteasomal degradation of Foxp3 through deubiquitination and stabilization of TAZ in vitro and in vivo.Notably,ML323,a specific inhibitor of the USP1/UAF1 deubiquitinase complex,inhibited Th17-cell differentiation and promoted Treg-cell differentiation in vitro and in vivo,indicating that ML323 might be a promising candidate for the treatment of diseases associated with an imbalance between Th17 and Treg cells.Our study highlights the critical role of USP1 in regulating adaptive immune responses and suggests that USP1 might be a drug target for the treatment of diseases associated with an imbalance between Th17 and Treg cells.展开更多
目的:研究泛素特异性肽酶1(ubiquitin specific peptidase 1,USP1)在卵巢癌中的表达及其与患者临床病理特征的相关性,探讨USP1对卵巢癌细胞恶性表型的影响。方法:生物信息学分析USP1在卵巢癌患者中的表达水平及与患者临床分期的关系;免...目的:研究泛素特异性肽酶1(ubiquitin specific peptidase 1,USP1)在卵巢癌中的表达及其与患者临床病理特征的相关性,探讨USP1对卵巢癌细胞恶性表型的影响。方法:生物信息学分析USP1在卵巢癌患者中的表达水平及与患者临床分期的关系;免疫组化检测USP1在卵巢癌患者和正常卵巢组织中的表达情况,分析其表达水平与患者临床特征之间的关系;通过慢病毒感染的方式构建过表达USP1的SKOV3细胞株,利用CCK-8实验和克隆形成实验检测细胞增殖和克隆形成能力,利用Transwell实验检测细胞迁移和侵袭能力。结果:卵巢癌组织中的USP1表达水平明显高于正常卵巢组织,并与卵巢癌患者病理类型、腹膜转移以及原位肿瘤的大小或侵犯程度相关,与Ki67、ER、PR、P53的表达水平不相关;体外实验结果表明USP1促进卵巢癌细胞增殖、克隆形成、迁移和侵袭能力。结论:USP1在卵巢癌组织中表达增强,并与患者病理类型、腹膜转移以及临床分期相关,USP1促进卵巢癌细胞恶性表型进展。展开更多
Objective:Small cell lung cancer(SCLC)is commonly recognized as the most fatal lung cancer type.Despite substantial advances in immune checkpoint blockade therapies for treating solid cancers,their benefits are limite...Objective:Small cell lung cancer(SCLC)is commonly recognized as the most fatal lung cancer type.Despite substantial advances in immune checkpoint blockade therapies for treating solid cancers,their benefits are limited to a minority of patients with SCLC.In the present study,novel indicators for predicting the outcomes and molecular targets for SCLC treatment were elucidated.Methods:We conducted bioinformatics analysis to identify the key genes associated with tumor-infiltrating lymphocytes in SCLC.The functional role of the key gene identified in SCLC was determined both in vitro and in vivo.Results:A significant correlation was observed between patient survival and CD56dim natural killer(NK)cell proportion.Furthermore,we noted that the hub gene ubiquitin-specific protease 1(USP1)is closely correlated with both CD56dim NK cells and overall survival in SCLC.Bioinformatics analysis revealed that USP1 is upregulated in SCLC.In addition,gene set enrichment analysis revealed that USP1 overexpression hinders NK cell-mediated immune responses.By co-cultivating NK-92 cells with SCLC cells,we demonstrated that NK cell cytotoxicity against SCLC could be improved either via USP1 knock-down or pharmacological inhibition.Furthermore,using a nude-mice xenograft tumor model,we noted that USP1 inhibition effectively suppressed tumor proliferation and increased the expression of NK cell-associated markers.Conclusions:Our study findings highlight the importance of NK cells in regulating SCLC.USP1 overexpression can inhibit NK cell-mediated immunity;therefore,USP1 may serve not only as a prognostic biomarker but also as a potential molecular target of SCLC therapy.展开更多
目的合成泛素特异性蛋白酶1(ubiquitin specific protease 1,USP1)抑制剂KSQ-4279。方法以4-甲酰苯甲酸甲酯为起始原料,经环合、N-烷基化、还原、氯代得到2-[4-(氯甲基)苯基]-1-异丙基-4-(三氟甲基)-1H-咪唑(6);以2,4-二氯嘧啶-5-甲酸...目的合成泛素特异性蛋白酶1(ubiquitin specific protease 1,USP1)抑制剂KSQ-4279。方法以4-甲酰苯甲酸甲酯为起始原料,经环合、N-烷基化、还原、氯代得到2-[4-(氯甲基)苯基]-1-异丙基-4-(三氟甲基)-1H-咪唑(6);以2,4-二氯嘧啶-5-甲酸甲酯为起始原料,经还原、氧化、卤代、关环得到6-氯-1H-吡唑并[3,4-d]嘧啶(10);中间体10与6发生N-烷基化后经Suzuki偶联反应得到目标化合物KSQ-4279。结果与结论合成的目标化合物经LC-MS和1H-NMR谱确证,纯度为98.10%(HPLC面积归一化法)。该路线反应条件温和,避免了危险化学试剂的使用,起始原料及催化剂价廉易得,更符合工业化生产要求。展开更多
[目的]探讨miR-340-5p对BRCA1缺陷型卵巢癌细胞复制叉稳定性的潜在机制。[方法]使用来自癌症基因组图谱(TCGA)卵巢癌数据集,将BRCA1突变肿瘤中的miR-340-5p表达与BRCA1野生型肿瘤进行比较。过表达或敲低miR-340-5p后,检测BRCA1缺陷型UWB...[目的]探讨miR-340-5p对BRCA1缺陷型卵巢癌细胞复制叉稳定性的潜在机制。[方法]使用来自癌症基因组图谱(TCGA)卵巢癌数据集,将BRCA1突变肿瘤中的miR-340-5p表达与BRCA1野生型肿瘤进行比较。过表达或敲低miR-340-5p后,检测BRCA1缺陷型UWB1.289细胞中复制叉的稳定性、细胞增殖水平。UWB1.289细胞中敲低或过表达miR-340-5p后进行转录组测序并使用siRNA筛选鉴定miR-340-5p的靶向基因。[结果]miR-340-5p低表达与乳腺肿瘤中BRCA1缺乏相关(0.04±0.01 vs 0.15±0.04,P<0.05)。具有低表达miR-340-5p的BRCA1缺陷型肿瘤具有更高水平的基因组改变(0.37±0.05 vs 0.66±0.11,P<0.05)。过表达miR-340-5p后,BRCA1缺陷型UWB1.289细胞中复制叉不稳定性增强(1.12±0.23 vs 0.81±0.10,P<0.05)。相反,在UWB1.289细胞中同时过表达BRCA1和miR-340-5p并没有增强复制叉的不稳定性。此外,过表达或者敲低miR-340-5p后UWB1.289细胞的增殖水平分别下降或者上升(P<0.05),而在过表达BRCA1的UWB1.289细胞中无此现象。荧光素酶报告实验发现miR-340-5p靶向USP1 mRNA的3'端非翻译区(P<0.05)。过表达或者敲低miR-340-5p后,USP1的mRNA和蛋白水平分别下降或者上升(P<0.05)。敲低USP1或者过表达USP1时,UWB1.289细胞中复制叉不稳定性增强(P<0.05)。同时,敲低USP1后,UWB1.289细胞的增殖水平下降(P<0.05);过表达USP1后UWB1.289细胞的增殖水平上升(P<0.05),而在过表达BRCA1的UWB1.289细胞中无此现象。[结论]miR-340-5p能够促进BRCA1缺陷卵巢癌细胞复制叉的不稳定性,并抑制BRCA1缺陷细胞的存活。展开更多
基金the National Natural Science Foundation of China(81971805,81901614,81901659,32070906,82072384,82074160,and 82272239).
文摘The balance between inflammatory T helper type 17(Th17)and immunosuppressive regulatory T(Treg)cells is critical for maintaining immune homeostasis in the human body and is tightly regulated under healthy conditions.An increasing number of studies have reported that deubiquitinases(DUBs)play a vital role in regulating Th17-and Treg-cell differentiation.However,the biological functions of only a small fraction of DUBs in Th17-and Treg-cell differentiation are well defined.In this study,we identified ubiquitin-specific peptidase 1(USP1)as a vital regulator of CD4+T-cell differentiation.USP1 promoted Th17-cell differentiation but attenuated Treg-cell differentiation,thereby promoting the development of inflammatory diseases.Mechanistically,USP1 in CD4+T cells enhanced the activity of RORγt but promoted the proteasomal degradation of Foxp3 through deubiquitination and stabilization of TAZ in vitro and in vivo.Notably,ML323,a specific inhibitor of the USP1/UAF1 deubiquitinase complex,inhibited Th17-cell differentiation and promoted Treg-cell differentiation in vitro and in vivo,indicating that ML323 might be a promising candidate for the treatment of diseases associated with an imbalance between Th17 and Treg cells.Our study highlights the critical role of USP1 in regulating adaptive immune responses and suggests that USP1 might be a drug target for the treatment of diseases associated with an imbalance between Th17 and Treg cells.
文摘目的:研究泛素特异性肽酶1(ubiquitin specific peptidase 1,USP1)在卵巢癌中的表达及其与患者临床病理特征的相关性,探讨USP1对卵巢癌细胞恶性表型的影响。方法:生物信息学分析USP1在卵巢癌患者中的表达水平及与患者临床分期的关系;免疫组化检测USP1在卵巢癌患者和正常卵巢组织中的表达情况,分析其表达水平与患者临床特征之间的关系;通过慢病毒感染的方式构建过表达USP1的SKOV3细胞株,利用CCK-8实验和克隆形成实验检测细胞增殖和克隆形成能力,利用Transwell实验检测细胞迁移和侵袭能力。结果:卵巢癌组织中的USP1表达水平明显高于正常卵巢组织,并与卵巢癌患者病理类型、腹膜转移以及原位肿瘤的大小或侵犯程度相关,与Ki67、ER、PR、P53的表达水平不相关;体外实验结果表明USP1促进卵巢癌细胞增殖、克隆形成、迁移和侵袭能力。结论:USP1在卵巢癌组织中表达增强,并与患者病理类型、腹膜转移以及临床分期相关,USP1促进卵巢癌细胞恶性表型进展。
基金supported by grants from the Dongguan Science and Technology of Social Development Program(No.20231800940192)the Talent Development Foundation of the First Dongguan Affiliated Hospital of Guangdong Medical University(No.PU2023002).
文摘Objective:Small cell lung cancer(SCLC)is commonly recognized as the most fatal lung cancer type.Despite substantial advances in immune checkpoint blockade therapies for treating solid cancers,their benefits are limited to a minority of patients with SCLC.In the present study,novel indicators for predicting the outcomes and molecular targets for SCLC treatment were elucidated.Methods:We conducted bioinformatics analysis to identify the key genes associated with tumor-infiltrating lymphocytes in SCLC.The functional role of the key gene identified in SCLC was determined both in vitro and in vivo.Results:A significant correlation was observed between patient survival and CD56dim natural killer(NK)cell proportion.Furthermore,we noted that the hub gene ubiquitin-specific protease 1(USP1)is closely correlated with both CD56dim NK cells and overall survival in SCLC.Bioinformatics analysis revealed that USP1 is upregulated in SCLC.In addition,gene set enrichment analysis revealed that USP1 overexpression hinders NK cell-mediated immune responses.By co-cultivating NK-92 cells with SCLC cells,we demonstrated that NK cell cytotoxicity against SCLC could be improved either via USP1 knock-down or pharmacological inhibition.Furthermore,using a nude-mice xenograft tumor model,we noted that USP1 inhibition effectively suppressed tumor proliferation and increased the expression of NK cell-associated markers.Conclusions:Our study findings highlight the importance of NK cells in regulating SCLC.USP1 overexpression can inhibit NK cell-mediated immunity;therefore,USP1 may serve not only as a prognostic biomarker but also as a potential molecular target of SCLC therapy.
文摘目的合成泛素特异性蛋白酶1(ubiquitin specific protease 1,USP1)抑制剂KSQ-4279。方法以4-甲酰苯甲酸甲酯为起始原料,经环合、N-烷基化、还原、氯代得到2-[4-(氯甲基)苯基]-1-异丙基-4-(三氟甲基)-1H-咪唑(6);以2,4-二氯嘧啶-5-甲酸甲酯为起始原料,经还原、氧化、卤代、关环得到6-氯-1H-吡唑并[3,4-d]嘧啶(10);中间体10与6发生N-烷基化后经Suzuki偶联反应得到目标化合物KSQ-4279。结果与结论合成的目标化合物经LC-MS和1H-NMR谱确证,纯度为98.10%(HPLC面积归一化法)。该路线反应条件温和,避免了危险化学试剂的使用,起始原料及催化剂价廉易得,更符合工业化生产要求。
文摘[目的]探讨miR-340-5p对BRCA1缺陷型卵巢癌细胞复制叉稳定性的潜在机制。[方法]使用来自癌症基因组图谱(TCGA)卵巢癌数据集,将BRCA1突变肿瘤中的miR-340-5p表达与BRCA1野生型肿瘤进行比较。过表达或敲低miR-340-5p后,检测BRCA1缺陷型UWB1.289细胞中复制叉的稳定性、细胞增殖水平。UWB1.289细胞中敲低或过表达miR-340-5p后进行转录组测序并使用siRNA筛选鉴定miR-340-5p的靶向基因。[结果]miR-340-5p低表达与乳腺肿瘤中BRCA1缺乏相关(0.04±0.01 vs 0.15±0.04,P<0.05)。具有低表达miR-340-5p的BRCA1缺陷型肿瘤具有更高水平的基因组改变(0.37±0.05 vs 0.66±0.11,P<0.05)。过表达miR-340-5p后,BRCA1缺陷型UWB1.289细胞中复制叉不稳定性增强(1.12±0.23 vs 0.81±0.10,P<0.05)。相反,在UWB1.289细胞中同时过表达BRCA1和miR-340-5p并没有增强复制叉的不稳定性。此外,过表达或者敲低miR-340-5p后UWB1.289细胞的增殖水平分别下降或者上升(P<0.05),而在过表达BRCA1的UWB1.289细胞中无此现象。荧光素酶报告实验发现miR-340-5p靶向USP1 mRNA的3'端非翻译区(P<0.05)。过表达或者敲低miR-340-5p后,USP1的mRNA和蛋白水平分别下降或者上升(P<0.05)。敲低USP1或者过表达USP1时,UWB1.289细胞中复制叉不稳定性增强(P<0.05)。同时,敲低USP1后,UWB1.289细胞的增殖水平下降(P<0.05);过表达USP1后UWB1.289细胞的增殖水平上升(P<0.05),而在过表达BRCA1的UWB1.289细胞中无此现象。[结论]miR-340-5p能够促进BRCA1缺陷卵巢癌细胞复制叉的不稳定性,并抑制BRCA1缺陷细胞的存活。
