BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by ...BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by inducing apoptosis.However,the mechanism by which matrine regulates antiapoptotic proteins in CRC remains unclear.AIM To identify apoptotic proteins from proteomics and investigate the role of matrine in impeding CRC apoptosis by regulating these proteins.METHODS Tumor and adjacent normal tissues were collected from 52 patients with CRC who underwent surgery between January and December 2021.Data-independent acquisition quantitative proteomic analysis was performed to identify differentially expressed apoptotic proteins.The selected apoptotic proteins were identified through their association with tumor-node-metastasis(TNM)stage and prognosis,then confirmed by immunohistochemical(IHC)staining in validation cohort.In vitro,the role of matrine or apoptotic proteins on cancer cells were analyzed.RESULTS Compared to normal tissues,88 anti-apoptotic proteins from proteomic results were selected.Among them,Shankassociated RH domain interactor(SHARPIN)was identified because of its relationship with TNM stage and overall survival in TCGA database.In the IHC-confirmed cohort,SHARPIN was highly expressed in CRC tissues and localized in the cytoplasm.Higher SHARPIN expression was associated with TNM stage,carbohydrate antigen 153 levels,and gross type compared to low expression.SHARPIN knockdown promoted apoptosis,significantly upregulated the expression of Bcl-2 associated agonist of cell death,Bcl-2 associated X protein,caspase 3,and caspase 8,and downregulated B-cell lymphoma-2(P<0.05).Importantly,matrine treatment promoted apoptosis and reversed the proliferation,invasion,and migration of CRC cells by repressing SHARPIN.CONCLUSION SHARPIN was identified as an upregulated anti-apoptotic protein in CRC,and matrine exhibited anticancer effects by downregulating its expression.Thus,matrine appears to be a promising drug for CRC.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 National Key Development Plan for Precision Medicine Research,No.2017YFC0910002.
文摘BACKGROUND The 5-year survival rate of patients with colorectal cancer(CRC)in China is only 56.9%,highlighting the need for new therapeutic drugs.Previous studies have shown that matrine exhibits antitumor effects by inducing apoptosis.However,the mechanism by which matrine regulates antiapoptotic proteins in CRC remains unclear.AIM To identify apoptotic proteins from proteomics and investigate the role of matrine in impeding CRC apoptosis by regulating these proteins.METHODS Tumor and adjacent normal tissues were collected from 52 patients with CRC who underwent surgery between January and December 2021.Data-independent acquisition quantitative proteomic analysis was performed to identify differentially expressed apoptotic proteins.The selected apoptotic proteins were identified through their association with tumor-node-metastasis(TNM)stage and prognosis,then confirmed by immunohistochemical(IHC)staining in validation cohort.In vitro,the role of matrine or apoptotic proteins on cancer cells were analyzed.RESULTS Compared to normal tissues,88 anti-apoptotic proteins from proteomic results were selected.Among them,Shankassociated RH domain interactor(SHARPIN)was identified because of its relationship with TNM stage and overall survival in TCGA database.In the IHC-confirmed cohort,SHARPIN was highly expressed in CRC tissues and localized in the cytoplasm.Higher SHARPIN expression was associated with TNM stage,carbohydrate antigen 153 levels,and gross type compared to low expression.SHARPIN knockdown promoted apoptosis,significantly upregulated the expression of Bcl-2 associated agonist of cell death,Bcl-2 associated X protein,caspase 3,and caspase 8,and downregulated B-cell lymphoma-2(P<0.05).Importantly,matrine treatment promoted apoptosis and reversed the proliferation,invasion,and migration of CRC cells by repressing SHARPIN.CONCLUSION SHARPIN was identified as an upregulated anti-apoptotic protein in CRC,and matrine exhibited anticancer effects by downregulating its expression.Thus,matrine appears to be a promising drug for CRC.
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。
