Objective:Gemcitabine combined with nab-paclitaxel therapy(GnP)represents first-line chemotherapy for advanced pancreatic ductal adenocarcinoma(PDAC).However,the efficacy of GnP is diminished due to chemotherapeutic r...Objective:Gemcitabine combined with nab-paclitaxel therapy(GnP)represents first-line chemotherapy for advanced pancreatic ductal adenocarcinoma(PDAC).However,the efficacy of GnP is diminished due to chemotherapeutic resistance induced by the tumor microenvironment(TME),the underlying mechanisms of which remain poorly understood.Methods:Clinical data from patients with PDAC who underwent GnP therapy were collected and neutrophil infiltration in tumor tissues was assessed.PDAC cell lines and a mouse model of PDAC were utilized to determine the mechanisms underlying GnP resistance and to focus on tumor-associated neutrophils and neutrophil extracellular traps(NETs).Results:GnP therapy recruited neutrophils to the TME,which resulted in the formation of NETs that contributed to therapeutic resistance in the PDAC murine model.The NET inhibitor,PAD4,enhanced the efficacy of GnP by suppressing tumor growth.Furthermore,GnP significantly upregulated CXCL8 secretion in GnP-resistant MIA PaCa-2 cells,which was mediated by increased expression of GPRC5A in PDAC cells.Screening of classic NET-derived molecules identified cell-free DNA(cfDNA)as a pleiotropic factor that promoted tumor cell proliferation and migration and thereby contributed to chemotherapeutic resistance.In vivo experiments revealed that the combination of GnP with si GPRC5A or DNase was more effective in reducing tumor growth and prolonging survival in PDAC-bearing mice than either treatment alone.Conclusion:The GPRC5A-CXCL8-NET-cfDNA axis has a critical role in the development of therapeutic resistance to GnP in PDAC.Targeting this axis may represent a promising strategy for overcoming GnP resistance and thereby enhancing the efficacy of chemotherapy in PDAC.展开更多
1998年,在人类口腔鳞癌细胞株UMSCC-22B中首次发现了GPRC5A(Gprotein-coupled receptor family C,member 5,group A)基因。后续的研究提示,GPRC5A是肺癌特异的抑癌基因,与长期慢性感染和吸烟等因素密切相关;而在乳腺癌和胃癌中,GPRC5A...1998年,在人类口腔鳞癌细胞株UMSCC-22B中首次发现了GPRC5A(Gprotein-coupled receptor family C,member 5,group A)基因。后续的研究提示,GPRC5A是肺癌特异的抑癌基因,与长期慢性感染和吸烟等因素密切相关;而在乳腺癌和胃癌中,GPRC5A又作为癌基因促进肿瘤细胞增殖。GPRC5A受维甲酸、p53和环腺苷酸(cyclic adenosine monophosphate,cAMP)的调节,并活化非经典Wnt信号通路,抑制信号转导及转录活化因子3(signal transducers and activators of transcription 3,STAT3)信号通路。本综述重点阐述了GPRC5A的生物学特性,与肺癌、乳腺癌和胃癌发生的关系,及其潜在的信号调控通路,以期为肺癌、乳腺癌及胃癌的预防和靶向治疗提供新的思路。展开更多
目的:基于生物信息数据挖掘阐明G蛋白偶联受体C家族5A基因(G protein-coupled receptor familyC, m e m b e r5, g r o u pA, G P R C 5 A)在胰腺癌中的表达及临床意义。方法:检索O n c o m i n e,TCGA,HumanProteinAtlas等基因数据库,...目的:基于生物信息数据挖掘阐明G蛋白偶联受体C家族5A基因(G protein-coupled receptor familyC, m e m b e r5, g r o u pA, G P R C 5 A)在胰腺癌中的表达及临床意义。方法:检索O n c o m i n e,TCGA,HumanProteinAtlas等基因数据库,分析GPRC5A在胰腺癌中的表达差异,并分析其在不同肿瘤中的表达。采用Western印迹技术在武汉大学人民医院小样本队列中验证GPRC5A在胰腺癌及癌旁组织中的蛋白表达水平,采用Kaplan-Meier进行患者生存分析,并对GPRC5A与靶向药物敏感性关系进行分析。结果:对Oncomine数据库中有差异表达的295项研究数据进行差异性分析,发现胰腺癌组织GPRC5A基因的表达明显高于正常胰腺组织。对GPRC5A在不同肿瘤组织中表达的4184项研究进行荟萃分析,发现GPRC5A在胰腺癌组织中显著表达增高。通过生存分析发现高表达GPRC5A胰腺癌患者生存期明显低于低表达者,高表达患者预后更差。此外,GPRC5A表达与靶向药物厄洛替尼的药物敏感性有一定关联。结论:GPRC5A在胰腺癌组织中呈现高表达,与患者预后显著相关,其有可能成为胰腺癌诊断及药物治疗的新靶点。展开更多
目的通过检测G蛋白偶联受体家族C群5成员A(G protein coupled receptor C type group 5 member A,GPRC5A)在肝癌组织及细胞中的表达,探讨GPRC5A对肝癌细胞增殖、凋亡、氧化应激的影响及其相关作用机制。方法收集2018年12月~2019年11月...目的通过检测G蛋白偶联受体家族C群5成员A(G protein coupled receptor C type group 5 member A,GPRC5A)在肝癌组织及细胞中的表达,探讨GPRC5A对肝癌细胞增殖、凋亡、氧化应激的影响及其相关作用机制。方法收集2018年12月~2019年11月在解放军联勤保障部队第九六九医院行手术治疗的38例肝癌患者癌组织及配对癌旁正常组织样本;另选取人正常肝上皮细胞株HL-7702和人肝癌细胞株(HepG2,HCCLM3,HuH-7和MHCC-97H)进行研究。采用实时荧光定量聚合酶链反应(qRT-PCR)检测肝癌组织、癌旁正常组织及肝癌细胞、正常肝上皮细胞中GPRC5A表达量。通过转染pcDNA3.1-GPRC5A质粒构建过表达GPRC5A肝癌细胞株,采用MTT实验和V-FITC/PI凋亡试剂盒分别检测过表达GPRC5A对肝癌细胞增殖、凋亡的影响。采用活性氧指示剂DCFH-DA检测细胞中氧化应激相关因子活性氧(ROS),NAD+/NADH和三磷酸腺苷(ATP)水平。采用Western blot实验检测凋亡相关蛋白caspase-3及上皮生长因子(VEGF)蛋白表达水平。结果肝癌组织中GPRC5A表达较癌旁正常组织显著降低(0.34±0.09 vs 0.73±0.10),差异有统计学意义(t=17.869,P<0.01)。肝癌细胞HepG2(0.35±0.06),HCCLM3(0.38±0.10),HuH-7(0.53±0.07),MHCC-97H(0.67±0.06)中GPRC5A表达量显著低于人正常肝上皮细胞HL-7702中的GPRC5A表达量(1.00±0.08),差异有统计学意义(t=5.716~11.258,均P<0.05)。pcDNA3.1-GPRC5A组转染36 h细胞增殖吸光度(A)值(0.94±0.16)显著低于对照组(1.49±0.05)和pcDNA3.1组(1.45±0.07)(F=25.640,P<0.01)。GPRC5A过表达组VEGF(0.98±0.04)表达量较对照组(2.94±0.15)和pcDNA3.1组(2.89±0.11)显著降低(F=310.450,P<0.001)。pcDNA3.1-GPRC5A组细胞中ROS(182.12±13.42)水平显著高于对照组(101.23±11.20)和pcDNA3.1组(98.30±10.26)(F=49.577,P<0.001);NAD+/NADH(35.24±6.43)及ATP(55.34±6.51)水平显著低于对照组(100.25±12.41,100.17±14.36)和pcDNA3.1组(97.86±9.67,102.23±11.02)(F=42.338,17.077,P<0.05)。GPRC5A过表达组细胞凋亡率高于对照组和pcDNA3.1组;细胞凋亡蛋白caspase-3(2.61±0.16)表达量也高于对照组(1.00±0.11)和pcDNA3.1组(1.01±0.02)(F=202.843,P<0.001)。pcDNA3.1-GPRC5A组细胞中p-STAT3表达量(0.43±0.06)显著低于对照组(1.00±0.13)和pcDNA3.1组(1.03±0.12)(F=29.476,P<0.001);pcDNA3.1-GPRC5A组下游基因Socs3(0.47±0.05),c-MYC(0.54±0.06)表达量也显著低于对照组(1.01±0.05,1.00±0.04)和pcDNA3.1组(0.98±0.09,1.02±0.06)(F=63.275,75.409,P<0.001)。肝癌组织中STAT3与GPRC5A表达量呈显著负相关(r=-0.746,P<0.01)。过转染pcDNA3.1-STAT3或pcDNA3.1-NLRP3后显著逆转了pcDNA3.1-GPRC5A对肝癌细胞的抑制作用(P<0.05)。结论GPRC5A低表达可能通过调节STAT3/Socs3/c-MYC信号和炎症反应抑制了肝癌细胞的增殖,诱导肝癌细胞的氧化应激和凋亡。展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.82271883 and 82073195 to Qing.Xiaand 82471775 and 82271800 to Yuli.Lin)+3 种基金the Hospital-pharma Integration Project on Innovative Achievement Translation(Grant No.SHDC2022CRD043 to Qing.Xia)the Shanghai Jiao Tong University School of Medicine Summit Program-“Research Physician”in Clinical Medicine to Qing.Xiathe Young Physician-Scientist Cultivation Program of Shanghai Immune Therapy Institute to Qing.Xiathe Light of the Lintel-young Talents of Huangpu to Qing.Xia。
文摘Objective:Gemcitabine combined with nab-paclitaxel therapy(GnP)represents first-line chemotherapy for advanced pancreatic ductal adenocarcinoma(PDAC).However,the efficacy of GnP is diminished due to chemotherapeutic resistance induced by the tumor microenvironment(TME),the underlying mechanisms of which remain poorly understood.Methods:Clinical data from patients with PDAC who underwent GnP therapy were collected and neutrophil infiltration in tumor tissues was assessed.PDAC cell lines and a mouse model of PDAC were utilized to determine the mechanisms underlying GnP resistance and to focus on tumor-associated neutrophils and neutrophil extracellular traps(NETs).Results:GnP therapy recruited neutrophils to the TME,which resulted in the formation of NETs that contributed to therapeutic resistance in the PDAC murine model.The NET inhibitor,PAD4,enhanced the efficacy of GnP by suppressing tumor growth.Furthermore,GnP significantly upregulated CXCL8 secretion in GnP-resistant MIA PaCa-2 cells,which was mediated by increased expression of GPRC5A in PDAC cells.Screening of classic NET-derived molecules identified cell-free DNA(cfDNA)as a pleiotropic factor that promoted tumor cell proliferation and migration and thereby contributed to chemotherapeutic resistance.In vivo experiments revealed that the combination of GnP with si GPRC5A or DNase was more effective in reducing tumor growth and prolonging survival in PDAC-bearing mice than either treatment alone.Conclusion:The GPRC5A-CXCL8-NET-cfDNA axis has a critical role in the development of therapeutic resistance to GnP in PDAC.Targeting this axis may represent a promising strategy for overcoming GnP resistance and thereby enhancing the efficacy of chemotherapy in PDAC.
文摘1998年,在人类口腔鳞癌细胞株UMSCC-22B中首次发现了GPRC5A(Gprotein-coupled receptor family C,member 5,group A)基因。后续的研究提示,GPRC5A是肺癌特异的抑癌基因,与长期慢性感染和吸烟等因素密切相关;而在乳腺癌和胃癌中,GPRC5A又作为癌基因促进肿瘤细胞增殖。GPRC5A受维甲酸、p53和环腺苷酸(cyclic adenosine monophosphate,cAMP)的调节,并活化非经典Wnt信号通路,抑制信号转导及转录活化因子3(signal transducers and activators of transcription 3,STAT3)信号通路。本综述重点阐述了GPRC5A的生物学特性,与肺癌、乳腺癌和胃癌发生的关系,及其潜在的信号调控通路,以期为肺癌、乳腺癌及胃癌的预防和靶向治疗提供新的思路。
文摘目的:基于生物信息数据挖掘阐明G蛋白偶联受体C家族5A基因(G protein-coupled receptor familyC, m e m b e r5, g r o u pA, G P R C 5 A)在胰腺癌中的表达及临床意义。方法:检索O n c o m i n e,TCGA,HumanProteinAtlas等基因数据库,分析GPRC5A在胰腺癌中的表达差异,并分析其在不同肿瘤中的表达。采用Western印迹技术在武汉大学人民医院小样本队列中验证GPRC5A在胰腺癌及癌旁组织中的蛋白表达水平,采用Kaplan-Meier进行患者生存分析,并对GPRC5A与靶向药物敏感性关系进行分析。结果:对Oncomine数据库中有差异表达的295项研究数据进行差异性分析,发现胰腺癌组织GPRC5A基因的表达明显高于正常胰腺组织。对GPRC5A在不同肿瘤组织中表达的4184项研究进行荟萃分析,发现GPRC5A在胰腺癌组织中显著表达增高。通过生存分析发现高表达GPRC5A胰腺癌患者生存期明显低于低表达者,高表达患者预后更差。此外,GPRC5A表达与靶向药物厄洛替尼的药物敏感性有一定关联。结论:GPRC5A在胰腺癌组织中呈现高表达,与患者预后显著相关,其有可能成为胰腺癌诊断及药物治疗的新靶点。
文摘目的通过检测G蛋白偶联受体家族C群5成员A(G protein coupled receptor C type group 5 member A,GPRC5A)在肝癌组织及细胞中的表达,探讨GPRC5A对肝癌细胞增殖、凋亡、氧化应激的影响及其相关作用机制。方法收集2018年12月~2019年11月在解放军联勤保障部队第九六九医院行手术治疗的38例肝癌患者癌组织及配对癌旁正常组织样本;另选取人正常肝上皮细胞株HL-7702和人肝癌细胞株(HepG2,HCCLM3,HuH-7和MHCC-97H)进行研究。采用实时荧光定量聚合酶链反应(qRT-PCR)检测肝癌组织、癌旁正常组织及肝癌细胞、正常肝上皮细胞中GPRC5A表达量。通过转染pcDNA3.1-GPRC5A质粒构建过表达GPRC5A肝癌细胞株,采用MTT实验和V-FITC/PI凋亡试剂盒分别检测过表达GPRC5A对肝癌细胞增殖、凋亡的影响。采用活性氧指示剂DCFH-DA检测细胞中氧化应激相关因子活性氧(ROS),NAD+/NADH和三磷酸腺苷(ATP)水平。采用Western blot实验检测凋亡相关蛋白caspase-3及上皮生长因子(VEGF)蛋白表达水平。结果肝癌组织中GPRC5A表达较癌旁正常组织显著降低(0.34±0.09 vs 0.73±0.10),差异有统计学意义(t=17.869,P<0.01)。肝癌细胞HepG2(0.35±0.06),HCCLM3(0.38±0.10),HuH-7(0.53±0.07),MHCC-97H(0.67±0.06)中GPRC5A表达量显著低于人正常肝上皮细胞HL-7702中的GPRC5A表达量(1.00±0.08),差异有统计学意义(t=5.716~11.258,均P<0.05)。pcDNA3.1-GPRC5A组转染36 h细胞增殖吸光度(A)值(0.94±0.16)显著低于对照组(1.49±0.05)和pcDNA3.1组(1.45±0.07)(F=25.640,P<0.01)。GPRC5A过表达组VEGF(0.98±0.04)表达量较对照组(2.94±0.15)和pcDNA3.1组(2.89±0.11)显著降低(F=310.450,P<0.001)。pcDNA3.1-GPRC5A组细胞中ROS(182.12±13.42)水平显著高于对照组(101.23±11.20)和pcDNA3.1组(98.30±10.26)(F=49.577,P<0.001);NAD+/NADH(35.24±6.43)及ATP(55.34±6.51)水平显著低于对照组(100.25±12.41,100.17±14.36)和pcDNA3.1组(97.86±9.67,102.23±11.02)(F=42.338,17.077,P<0.05)。GPRC5A过表达组细胞凋亡率高于对照组和pcDNA3.1组;细胞凋亡蛋白caspase-3(2.61±0.16)表达量也高于对照组(1.00±0.11)和pcDNA3.1组(1.01±0.02)(F=202.843,P<0.001)。pcDNA3.1-GPRC5A组细胞中p-STAT3表达量(0.43±0.06)显著低于对照组(1.00±0.13)和pcDNA3.1组(1.03±0.12)(F=29.476,P<0.001);pcDNA3.1-GPRC5A组下游基因Socs3(0.47±0.05),c-MYC(0.54±0.06)表达量也显著低于对照组(1.01±0.05,1.00±0.04)和pcDNA3.1组(0.98±0.09,1.02±0.06)(F=63.275,75.409,P<0.001)。肝癌组织中STAT3与GPRC5A表达量呈显著负相关(r=-0.746,P<0.01)。过转染pcDNA3.1-STAT3或pcDNA3.1-NLRP3后显著逆转了pcDNA3.1-GPRC5A对肝癌细胞的抑制作用(P<0.05)。结论GPRC5A低表达可能通过调节STAT3/Socs3/c-MYC信号和炎症反应抑制了肝癌细胞的增殖,诱导肝癌细胞的氧化应激和凋亡。
