Lung cancer ranks the top of malignancies that cause cancer-related deaths worldwide.The leaves of Morus alba L are traditional Chinese medicine widely applied in respiratory diseases.Our previous work has demonstrate...Lung cancer ranks the top of malignancies that cause cancer-related deaths worldwide.The leaves of Morus alba L are traditional Chinese medicine widely applied in respiratory diseases.Our previous work has demonstrated the anti-lung cancer effect of secondary metabolites of mulberry leaf,but their mechanism of action has still not fully elucidated.We synthesized Moracin N(MAN)-Probe conjugated with alkyne to label lung cancer cells and identified protein targets by chemical proteomic analysis.MAN and its probe exerted similar growth-inhibitory effect on human lung cancer cells.Chemical proteomic results showed that MAN targeted the programmed death ligand 1(PD-L1)checkpoint pathway and T cell receptor(TCR)signaling pathway,indicating its immune-regulatory function.Cell-free surface plasmon resonance(SPR)results showed the direct interaction of MAN with PD-L1 protein.Molecular docking analysis demonstrated that MAN bound to E158 residue of PD-L1 protein.MAN downregulated the expression levels of PD-L1 in a time-and dose-dependent manner and disrupted the PD-L1/programmed death 1(PD-1)binding,including other secondary metabolites of mulberry leaves Guangsangon E(GSE)and Chalcomoracin(CMR).Human peripheral blood mononuclear cells(PBMCs)co-cultured with MAN-treated A549 cells,resulting in the increase of CD8^(+)GZMB^(+)T cells and the decrease of CD8^(+)PD-1^(+)T cells.It suggested that MAN exerts anti-cancer effect through blocking the PD-L1/PD-1 signaling.In vivo,MAN combined with anti-PD-1 antibody significantly inhibited lung cancer development and metastasis,indicating their synergistic effect.Taken together,secondary metabolites of mulberry leaves target the PD-L1/PD-1 signaling,enhance T cell-mediated immunity and inhibit the tumorigenesis of lung cancer.Their modulatory effect on tumor microenvironment makes them able to enhance the therapeutic efficacy of immune checkpoint inhibitors in lung cancer.展开更多
程序性死亡受体1(programmed death 1,PD-1)和程序性死亡配体1(PD-1 ligand,PD-L1)共同组成T细胞介导的适应性免疫过程中的协同刺激信号通路。肿瘤发生时,T细胞表面的PD-1受体与肿瘤细胞表面表达的PD-1配体结合,通过抑制T细胞活性及其...程序性死亡受体1(programmed death 1,PD-1)和程序性死亡配体1(PD-1 ligand,PD-L1)共同组成T细胞介导的适应性免疫过程中的协同刺激信号通路。肿瘤发生时,T细胞表面的PD-1受体与肿瘤细胞表面表达的PD-1配体结合,通过抑制T细胞活性及其增殖、分化影响肿瘤细胞对常规治疗的耐受及肿瘤患者远期预后。据报道PD-L1在宫颈癌等多种肿瘤中过高表达,提示PD-1/PD-L1通路可能参与肿瘤的发生、发展及转归,因此阻断该通路或许成为肿瘤靶向治疗的一个具有划时代意义的创新。本文就PD-1/PD-L1信号通路的作用机制和与宫颈癌之间的关系进行综述,探讨该通路相关的免疫疗法在宫颈癌临床治疗中的意义。展开更多
背景众多证据表明免疫逃逸在肿瘤形成过程中扮演重要角色,慢性萎缩性胃炎(chronic atrophic gastritis,CAG)是胃癌的癌前疾病.安胃汤被发现可改善CAG临床症状及病理表现,实现CAG的逆转,该作用是否与免疫逃逸机制相关有待进一步研究.目...背景众多证据表明免疫逃逸在肿瘤形成过程中扮演重要角色,慢性萎缩性胃炎(chronic atrophic gastritis,CAG)是胃癌的癌前疾病.安胃汤被发现可改善CAG临床症状及病理表现,实现CAG的逆转,该作用是否与免疫逃逸机制相关有待进一步研究.目的从细胞免疫逃逸角度,探讨程序性死亡受体-1(programmed cell death protein 1,PD-1)/程序性死亡受体配体-1(programmed cell death ligand 1,PD-L1)信号轴与安胃汤对CAG模型大鼠疗效之间的关系.方法采用1-甲基-3-硝基-1-亚硝基胍(1-Methyl-3-nitro-1-nitrosoguanidine,MNNG)慢性萎缩性胃炎大鼠模型,应用不同剂量安胃汤及维酶素片进行干预;HE染色观察安胃汤对CAG模型大鼠胃黏膜炎症细胞浸润及组织形态改变的影响;免疫组化检测CAG模型大鼠胃黏膜组织PD-1、PD-L1蛋白表达;ELISA检测血清CD4^(+)、CD8^(+)水平变化;qPCR检测CAG模型大鼠胃黏膜PD-1mRNA、PD-L1mRNA表达;Western-blot检测CAG模型大鼠胃黏膜组织PD-1、PD-L1蛋白表达.结果免疫组化结果示:与模型组和维酶素组比较,安胃汤高、低剂量组PD-L1表达均较低(P<0.01,P<0.05).ELISA实验结果示:与模型组比较,安胃汤高剂量组CD4^(+)表达及CD4^(+)/CD8^(+)比值升高(P<0.01,P<0.05),安胃汤各组和维酶素组CD8^(+)表达降低(P<0.01);与维酶素组比较,安胃汤高剂量组CD8^(+)表达降低(P<0.05).qPCR实验结果显示:与模型组比较,安胃汤高剂量组和维酶素组PD-1mRNA表达下降(P<0.01),安胃汤高、中剂量组PD-L1mRNA表达下降(P<0.01,P<0.05).Western-blot实验结果显示:与模型组比较,安胃汤高、中剂量组PD-1/Actin,PD-L1/Actin表达下降(P<0.01,P<0.05).结论安胃汤抗CAG作用可能与抑制PD-1/PD-L1信号通路诱导的细胞免疫逃逸有关.展开更多
目的探讨肉瘤样肾细胞癌组织中PD-L1的表达及肿瘤内微血管密度情况,为肉瘤样肾细胞癌免疫治疗及靶向治疗方案的选择提供理论依据。方法通过免疫组化法检测PD-L1、CD31及CD34在16例肉瘤样肾细胞癌(癌成分均为透明细胞肾细胞癌)中的表达,...目的探讨肉瘤样肾细胞癌组织中PD-L1的表达及肿瘤内微血管密度情况,为肉瘤样肾细胞癌免疫治疗及靶向治疗方案的选择提供理论依据。方法通过免疫组化法检测PD-L1、CD31及CD34在16例肉瘤样肾细胞癌(癌成分均为透明细胞肾细胞癌)中的表达,并评估肿瘤微血管密度。结果16例肿瘤中CD31和CD34免疫组化染色显示,肉瘤样肾细胞癌区域微血管密度明显高于不伴肉瘤样分化的区域,微血管密度计数分别为68.6±25.8 vs 38.7±16.0(t=3.931,P=0.0005)和69.5±28.1 vs 40.1±18.4(t=3.506,P=0.0015),差异有统计学意义。肉瘤样区域PD-L1表达水平高于非肉瘤样区域,CPS分别为34.7±26.9和25.9±27.6,但差异无统计学意义。结论在肉瘤样肾细胞癌中,肉瘤样区域微血管密度和PD-L1表达水平明显高于非肉瘤样区域,提示靶向治疗联合免疫治疗可能为此类肿瘤提供一种有效的治疗方法。展开更多
基金supported by the National Natural Science Foundation of China(Grant No.:32070740)Zhejiang Provincial Natural Science Foundation(Grant No.:LZ23H160005)+1 种基金Natural Science Foundation of Jiangsu Province(Grant No.:BK20201197)Zhejiang Provincial Outstanding Talent Project of Ten Thousand Talents Program,Zhejiang Provincial Qianjiang Talents Program to Jianbin Zhang.
文摘Lung cancer ranks the top of malignancies that cause cancer-related deaths worldwide.The leaves of Morus alba L are traditional Chinese medicine widely applied in respiratory diseases.Our previous work has demonstrated the anti-lung cancer effect of secondary metabolites of mulberry leaf,but their mechanism of action has still not fully elucidated.We synthesized Moracin N(MAN)-Probe conjugated with alkyne to label lung cancer cells and identified protein targets by chemical proteomic analysis.MAN and its probe exerted similar growth-inhibitory effect on human lung cancer cells.Chemical proteomic results showed that MAN targeted the programmed death ligand 1(PD-L1)checkpoint pathway and T cell receptor(TCR)signaling pathway,indicating its immune-regulatory function.Cell-free surface plasmon resonance(SPR)results showed the direct interaction of MAN with PD-L1 protein.Molecular docking analysis demonstrated that MAN bound to E158 residue of PD-L1 protein.MAN downregulated the expression levels of PD-L1 in a time-and dose-dependent manner and disrupted the PD-L1/programmed death 1(PD-1)binding,including other secondary metabolites of mulberry leaves Guangsangon E(GSE)and Chalcomoracin(CMR).Human peripheral blood mononuclear cells(PBMCs)co-cultured with MAN-treated A549 cells,resulting in the increase of CD8^(+)GZMB^(+)T cells and the decrease of CD8^(+)PD-1^(+)T cells.It suggested that MAN exerts anti-cancer effect through blocking the PD-L1/PD-1 signaling.In vivo,MAN combined with anti-PD-1 antibody significantly inhibited lung cancer development and metastasis,indicating their synergistic effect.Taken together,secondary metabolites of mulberry leaves target the PD-L1/PD-1 signaling,enhance T cell-mediated immunity and inhibit the tumorigenesis of lung cancer.Their modulatory effect on tumor microenvironment makes them able to enhance the therapeutic efficacy of immune checkpoint inhibitors in lung cancer.
文摘程序性死亡受体1(programmed death 1,PD-1)和程序性死亡配体1(PD-1 ligand,PD-L1)共同组成T细胞介导的适应性免疫过程中的协同刺激信号通路。肿瘤发生时,T细胞表面的PD-1受体与肿瘤细胞表面表达的PD-1配体结合,通过抑制T细胞活性及其增殖、分化影响肿瘤细胞对常规治疗的耐受及肿瘤患者远期预后。据报道PD-L1在宫颈癌等多种肿瘤中过高表达,提示PD-1/PD-L1通路可能参与肿瘤的发生、发展及转归,因此阻断该通路或许成为肿瘤靶向治疗的一个具有划时代意义的创新。本文就PD-1/PD-L1信号通路的作用机制和与宫颈癌之间的关系进行综述,探讨该通路相关的免疫疗法在宫颈癌临床治疗中的意义。
文摘背景众多证据表明免疫逃逸在肿瘤形成过程中扮演重要角色,慢性萎缩性胃炎(chronic atrophic gastritis,CAG)是胃癌的癌前疾病.安胃汤被发现可改善CAG临床症状及病理表现,实现CAG的逆转,该作用是否与免疫逃逸机制相关有待进一步研究.目的从细胞免疫逃逸角度,探讨程序性死亡受体-1(programmed cell death protein 1,PD-1)/程序性死亡受体配体-1(programmed cell death ligand 1,PD-L1)信号轴与安胃汤对CAG模型大鼠疗效之间的关系.方法采用1-甲基-3-硝基-1-亚硝基胍(1-Methyl-3-nitro-1-nitrosoguanidine,MNNG)慢性萎缩性胃炎大鼠模型,应用不同剂量安胃汤及维酶素片进行干预;HE染色观察安胃汤对CAG模型大鼠胃黏膜炎症细胞浸润及组织形态改变的影响;免疫组化检测CAG模型大鼠胃黏膜组织PD-1、PD-L1蛋白表达;ELISA检测血清CD4^(+)、CD8^(+)水平变化;qPCR检测CAG模型大鼠胃黏膜PD-1mRNA、PD-L1mRNA表达;Western-blot检测CAG模型大鼠胃黏膜组织PD-1、PD-L1蛋白表达.结果免疫组化结果示:与模型组和维酶素组比较,安胃汤高、低剂量组PD-L1表达均较低(P<0.01,P<0.05).ELISA实验结果示:与模型组比较,安胃汤高剂量组CD4^(+)表达及CD4^(+)/CD8^(+)比值升高(P<0.01,P<0.05),安胃汤各组和维酶素组CD8^(+)表达降低(P<0.01);与维酶素组比较,安胃汤高剂量组CD8^(+)表达降低(P<0.05).qPCR实验结果显示:与模型组比较,安胃汤高剂量组和维酶素组PD-1mRNA表达下降(P<0.01),安胃汤高、中剂量组PD-L1mRNA表达下降(P<0.01,P<0.05).Western-blot实验结果显示:与模型组比较,安胃汤高、中剂量组PD-1/Actin,PD-L1/Actin表达下降(P<0.01,P<0.05).结论安胃汤抗CAG作用可能与抑制PD-1/PD-L1信号通路诱导的细胞免疫逃逸有关.
文摘目的探讨肉瘤样肾细胞癌组织中PD-L1的表达及肿瘤内微血管密度情况,为肉瘤样肾细胞癌免疫治疗及靶向治疗方案的选择提供理论依据。方法通过免疫组化法检测PD-L1、CD31及CD34在16例肉瘤样肾细胞癌(癌成分均为透明细胞肾细胞癌)中的表达,并评估肿瘤微血管密度。结果16例肿瘤中CD31和CD34免疫组化染色显示,肉瘤样肾细胞癌区域微血管密度明显高于不伴肉瘤样分化的区域,微血管密度计数分别为68.6±25.8 vs 38.7±16.0(t=3.931,P=0.0005)和69.5±28.1 vs 40.1±18.4(t=3.506,P=0.0015),差异有统计学意义。肉瘤样区域PD-L1表达水平高于非肉瘤样区域,CPS分别为34.7±26.9和25.9±27.6,但差异无统计学意义。结论在肉瘤样肾细胞癌中,肉瘤样区域微血管密度和PD-L1表达水平明显高于非肉瘤样区域,提示靶向治疗联合免疫治疗可能为此类肿瘤提供一种有效的治疗方法。
