Doxorubicin (Dox) is a major anticancer chemotherapeutic agent. However, it causes cardiomyopathy due to the side effect of cardiomyocyte apoptosis. We have previously reported that angiopoietin-1 significantly redu...Doxorubicin (Dox) is a major anticancer chemotherapeutic agent. However, it causes cardiomyopathy due to the side effect of cardiomyocyte apoptosis. We have previously reported that angiopoietin-1 significantly reduced myocardial infarction after ischemic injury and protected cardiomyocytes from oxidative stress-induced apoptosis. It is hypothesized that angiopoietin-1 may protect cardiomyocytes from Dox-induced apoptosis. Cardiomyocytes H9C2 were transfected with adenovirus expressing angiopoietin-1 (Ad5-Ang-1) 24 h before the cells were chal- lenged with Dox at a concentration of 2 ~tmol/L. Ad5-GFP served as the vector control. Cardiomyocyte apoptosis was evaluated using Annexin V-FITC staining and caspase-3 and caspase-8 activity was determined by Western blotting. The results showed that Dox treatment significantly induced cardiomyocyte apoptosis as evidenced by the greater number of Annexin V-FITC stained cells and increases in caspase-3 and caspase-8 activity. In contrast, overexpression of angiopoietin-1 significantly prevented Dox-induced cardiomyocyte apoptosis. To elucidate the mechanisms by which angiopoietin-1 protected cells from Dox-induced apoptosis, we analyzed both extrinsic and intrinsic apoptotic signaling pathways. We observed that angiopoietin-1 prevented Dox-induced activation of both extrinsic and intrinsic apoptotic signaling pathways. Specifically, angiopoietin-1 prevented DOX-induced in- creases in FasL and Bax levels and cleaved caspase-3 and caspase-8 levels in H9C2 cells. In addition, overexpres- sion of angiopoietin-1 also activated the pro-survival phosphoinositide-3 kinase (PI3K)/Akt signaling pathway and decreased Dox-induced nuclear factor-kappaB (NF-~:B) activation. Our data suggest that promoting the expression of angiopoietin-1 could be a potential approach for reducing Dox-induced cardiomyocyte cytoxicity.展开更多
目的:探究钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)在肺腺癌组织中的表达及其促进肺腺癌的侵袭、转移。方法:通过免疫组织化学染色(immunohistochemistry,IHC)分析肺腺癌患者的石蜡组织标本...目的:探究钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)在肺腺癌组织中的表达及其促进肺腺癌的侵袭、转移。方法:通过免疫组织化学染色(immunohistochemistry,IHC)分析肺腺癌患者的石蜡组织标本CaMKⅡ表达与临床病理参数关系,同时对肺腺癌组织与其配对淋巴结转移病灶中的CaMKⅡ表达情况进行比较分析,收集2011年1月至2011年12月于天津医科大学肿瘤医院行手术治疗的113例肺腺癌患者及21例配对的原发病灶及淋巴结转移病灶的石蜡组织标本。将肺腺癌细胞系H1299及Calu3进行慢病毒转染,实验组转染高表达CaMKⅡ病毒,对照组转染阴性对照病毒,通过Transwell实验和划痕实验检测肺腺癌细胞侵袭、转移能力,通过Western blot检测CaMKⅡ表达水平与上皮间充质转化(pithelial-mesenchymal transition,EMT)相关指标和表皮生长因子受体(epidermal growth factor receptor,EGFR)通路激活间的关系。结果:CaMKⅡ高表达与肺腺癌TNM分期和淋巴结转移呈正相关,肺腺癌淋巴结转移病灶中的癌组织CaMKⅡ表达明显高于其原发病灶(P<0.05)。细胞实验表明,CaMKⅡ高表达的肺腺癌细胞,其穿膜细胞数目明显增多,伤口愈合能力增强;EGFR通路激活后p-CaMKⅡ水平增加,且CaMKⅡ促进了EMT相关蛋白及转录因子的表达。结论:CaMKⅡ参与EGFR信号传导,促进EMT过程,增加肺腺癌的侵袭转移能力。展开更多
Objective:The immunoscore,which is used to quantify immune infiltrates,has greater relative prognostic value than tumor,node,and metastasis(TNM)stage and might serve as a new system for classification of colorectal ca...Objective:The immunoscore,which is used to quantify immune infiltrates,has greater relative prognostic value than tumor,node,and metastasis(TNM)stage and might serve as a new system for classification of colorectal cancer.However,a comparable immunoscore for predicting lung adenocarcinoma(LUAD)prognosis is currently lacking.Methods:We analyzed the expression of 18 immune features by immunohistochemistry in 171 specimens.The relationship of immune marker expression and clinicopathologic factors to the overall survival(OS)was analyzed with the Kaplan-Meier method.A nomogram was developed by using the optimal features selected by least absolute shrinkage and selection operator(LASSO)regression in the training cohort(n=111)and evaluated in the validation cohort(n=60).Results:The indicators integrated in the nomogram were TNM stage,neuron-specific enolase,carcino-embryonic antigen,CD8 center of tumor(CT),CD8 invasive margin(IM),Fox P3 CT,and CD45 ROCT.The calibration curve showed prominent agreement between the observed 2-and 5-year OS and that predicted by the nomogram.To simplify the nomogram,we developed a new immune-serum scoring system(I-SSS)based on the points awarded for each factor in the nomogram.Our I-SSS was able to stratify same-stage patients into different risk subgroups.The combination of I-SSS and TNM stage had better prognostic value than the TNM stage alone.Conclusions:Our new I-SSS can accurately and individually predict LUAD prognosis and may be used to supplement prognostication based on the TNM stage.展开更多
基金supported by the National Natural Science Foundation of China (No. 30971258)the National 973 project of China(NO.2012CB517503)+1 种基金the Key Project of Natural Science Foundation of Jiangsu Province (No. 11KJA310004)Jiangsu "Six Personnel Peak" Talent-Funded Projects
文摘Doxorubicin (Dox) is a major anticancer chemotherapeutic agent. However, it causes cardiomyopathy due to the side effect of cardiomyocyte apoptosis. We have previously reported that angiopoietin-1 significantly reduced myocardial infarction after ischemic injury and protected cardiomyocytes from oxidative stress-induced apoptosis. It is hypothesized that angiopoietin-1 may protect cardiomyocytes from Dox-induced apoptosis. Cardiomyocytes H9C2 were transfected with adenovirus expressing angiopoietin-1 (Ad5-Ang-1) 24 h before the cells were chal- lenged with Dox at a concentration of 2 ~tmol/L. Ad5-GFP served as the vector control. Cardiomyocyte apoptosis was evaluated using Annexin V-FITC staining and caspase-3 and caspase-8 activity was determined by Western blotting. The results showed that Dox treatment significantly induced cardiomyocyte apoptosis as evidenced by the greater number of Annexin V-FITC stained cells and increases in caspase-3 and caspase-8 activity. In contrast, overexpression of angiopoietin-1 significantly prevented Dox-induced cardiomyocyte apoptosis. To elucidate the mechanisms by which angiopoietin-1 protected cells from Dox-induced apoptosis, we analyzed both extrinsic and intrinsic apoptotic signaling pathways. We observed that angiopoietin-1 prevented Dox-induced activation of both extrinsic and intrinsic apoptotic signaling pathways. Specifically, angiopoietin-1 prevented DOX-induced in- creases in FasL and Bax levels and cleaved caspase-3 and caspase-8 levels in H9C2 cells. In addition, overexpres- sion of angiopoietin-1 also activated the pro-survival phosphoinositide-3 kinase (PI3K)/Akt signaling pathway and decreased Dox-induced nuclear factor-kappaB (NF-~:B) activation. Our data suggest that promoting the expression of angiopoietin-1 could be a potential approach for reducing Dox-induced cardiomyocyte cytoxicity.
基金supported by the Tianjin Municipal Health Bureau Science and Technology Foundation(Grant No.16KG125)the Project of the National Natural Science Foundation of China(Grant No.81801781)the Project of Tumor Translational Medicine Seed Fund of the Tianjin Medical University Cancer Institute and Hospital(Grant No.1905)。
文摘Objective:The immunoscore,which is used to quantify immune infiltrates,has greater relative prognostic value than tumor,node,and metastasis(TNM)stage and might serve as a new system for classification of colorectal cancer.However,a comparable immunoscore for predicting lung adenocarcinoma(LUAD)prognosis is currently lacking.Methods:We analyzed the expression of 18 immune features by immunohistochemistry in 171 specimens.The relationship of immune marker expression and clinicopathologic factors to the overall survival(OS)was analyzed with the Kaplan-Meier method.A nomogram was developed by using the optimal features selected by least absolute shrinkage and selection operator(LASSO)regression in the training cohort(n=111)and evaluated in the validation cohort(n=60).Results:The indicators integrated in the nomogram were TNM stage,neuron-specific enolase,carcino-embryonic antigen,CD8 center of tumor(CT),CD8 invasive margin(IM),Fox P3 CT,and CD45 ROCT.The calibration curve showed prominent agreement between the observed 2-and 5-year OS and that predicted by the nomogram.To simplify the nomogram,we developed a new immune-serum scoring system(I-SSS)based on the points awarded for each factor in the nomogram.Our I-SSS was able to stratify same-stage patients into different risk subgroups.The combination of I-SSS and TNM stage had better prognostic value than the TNM stage alone.Conclusions:Our new I-SSS can accurately and individually predict LUAD prognosis and may be used to supplement prognostication based on the TNM stage.