目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁...目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁正常组织(P<0.05)。结论:PGAM1在乳腺癌组织中有高表达,为乳腺癌的诊断及治疗提供了新的思路。展开更多
Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality,necessitating novel therapeutic targets.This study explores the oncogenic role of integrin-linked kinase-associated phosphatase(ILKAP)in HCC ...Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality,necessitating novel therapeutic targets.This study explores the oncogenic role of integrin-linked kinase-associated phosphatase(ILKAP)in HCC and its underlying mechanisms.Database analyses(TCGA,UALCAN)revealed ILKAP overexpression in HCC,correlating with poor prognosis.Functional assays demonstrated that ILKAP knockdown significantly suppressed HCC cell proliferation and migration in vitro,while xenograft models confirmed its role in tumor growth in vivo.RNA sequencing identified 357 differentially expressed genes(DEGs),including 48 protein-coding DEGs,with glycolytic enzyme PGAM1 notably downregulated upon ILKAP silencing.ILKAP and PGAM1 expression were positively correlated in HCC tissues,and elevated PGAM1 levels were linked to worse survival.Notably,restoring PGAM1 in ILKAP-knockdown cells rescued proliferation and invasion,underscoring PGAM1’s critical role in ILKAP-mediated tumor progression.ILKAP depletion also reduced extracellular acidification rates and altered glycolysis-related gene expression,highlighting its role in metabolic reprogramming.These findings suggest that ILKAP drives HCC malignancy by modulating PGAM1 and glycolysis,providing a potential therapeutic target for HCC treatment.Further elucidation of the ILKAP-PGAM1 axis may offer new strategies for liver cancer management.展开更多
目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过...目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过表达组、miR-451a敲低组、温肺降浊方含药血清(WFJZF)组、miR-451a过表达+WFJZF组、miR-451a敲低+WFJZF组和miR-451a空载体组,双荧光素酶确定miR-451a和PGAM5的转录关系,免疫蛋白共沉淀确定PGAM5和Drp1的调控作用,慢病毒转染miR-451a于HT22细胞中,qRT-PCR确定转染效率,CCK-8测定缺糖缺氧状态下细胞活性的最佳时间点,qRT-PCR和Westem Blot检测HT22模型细胞内DGAM5-Drp1轴蛋白及mRNA的表达水平。结果:OGD/R诱导HT22细胞持续2 h为最佳时间点,miR-451a可以靶向调控PGAM5基因,PGAM5和Drp1具有相互调控作用。与正常对照组比较,OGD/R组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达上升,p-Drp1 Ser 616磷酸化表达上升(P<0.05),OPA1蛋白及mRNA表达下降,p-Drp1 Ser 637去磷酸化表达下降(P<0.05);与OGD/R组比较,miR-451a过表达+WFJZF组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达下降,p-Drp1 Ser 616磷酸化表达下降(P<0.05),OPA1蛋白及mRNA表达上升,p-Drp1 Ser 637去磷酸化表达上升(P<0.05)。结论:miR-451a可以靶向调控PGAM5-Drp1轴,miR-451a过表达+WFJZF组可以改善线粒体失衡状态,减少神经元过度损伤,发挥脑保护效应。展开更多
目的:探究磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)变构抑制剂能否提高结肠癌细胞对奥沙利铂(oxaliplatin,OXA)的敏感性。方法:癌症药物敏感性基因组学(genomics of drug sensitivity in cancer,GDSC)和癌细胞系百科全书数...目的:探究磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)变构抑制剂能否提高结肠癌细胞对奥沙利铂(oxaliplatin,OXA)的敏感性。方法:癌症药物敏感性基因组学(genomics of drug sensitivity in cancer,GDSC)和癌细胞系百科全书数据库(cancer cell line encyclopedia,CCLE)分析四种共识分子亚型(consensus molecular subgroups,CMS)结肠癌细胞对OXA敏感性和PGAM1的表达水平;采用CCK8法和Annexin V/PI双染法,探究PGAM1变构抑制剂HKB99对人源结肠癌细胞增殖和凋亡的作用;采用CCK8法分析HKB99联合OXA对结肠癌增殖的作用,并进行协同系数分析。结果:首先,GDSC和CCLE数据库分析结果显示,相对于CMS2和CMS3亚型,CMS1和CMS4亚型结肠癌细胞对OXA敏感的细胞比例降低,PGAM1表达升高,具有显著性差异(P<0.05)。进一步研究表明,HKB99显著抑制不同CMS类型的人源结肠癌SW480、SCUN2B和DLD-1细胞的增殖,其IC50均为1μM左右;Annexin V/PI双染法结果显示,与对照组相比,HKB99处理组DLD-1细胞的凋亡水平显著上升(P<0.01)。最后,CCK8法检测显示,与OXA单加药组相比,1.5μM的HKB99显著提高结肠癌DLD-1细胞对OXA的敏感性(IC_(50):3.26 vs0.37μM);协同系数分析结果显示,OXA在浓度1.56μM和25μM之间与HKB99的协同系数小于1。结论:本研究发现相对于CMS2和CMS3,CMS1和CMS4结肠癌细胞对OXA敏感性较低且PGAM1表达水平升高,PGAM1变构抑制剂HKB99增强结肠癌细胞对OXA的敏感性,提示HKB99有望提高结肠癌OXA化疗的敏感性,为患者个体化化疗提供新的治疗策略。展开更多
文摘目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁正常组织(P<0.05)。结论:PGAM1在乳腺癌组织中有高表达,为乳腺癌的诊断及治疗提供了新的思路。
基金supported by grants from the Natural Science Foundation of Zhejiang Province(No.LGF22H080016)the Key Research and Development Select Projects of Zhejiang Provincial Department of Science and Technology(No.2020C03008)+1 种基金the Zhejiang Medical Health Science and Technology Project(No.2022RC124)the Hangzhou Medical College Basal Research Fund(No.KYZD2024004).
文摘Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality,necessitating novel therapeutic targets.This study explores the oncogenic role of integrin-linked kinase-associated phosphatase(ILKAP)in HCC and its underlying mechanisms.Database analyses(TCGA,UALCAN)revealed ILKAP overexpression in HCC,correlating with poor prognosis.Functional assays demonstrated that ILKAP knockdown significantly suppressed HCC cell proliferation and migration in vitro,while xenograft models confirmed its role in tumor growth in vivo.RNA sequencing identified 357 differentially expressed genes(DEGs),including 48 protein-coding DEGs,with glycolytic enzyme PGAM1 notably downregulated upon ILKAP silencing.ILKAP and PGAM1 expression were positively correlated in HCC tissues,and elevated PGAM1 levels were linked to worse survival.Notably,restoring PGAM1 in ILKAP-knockdown cells rescued proliferation and invasion,underscoring PGAM1’s critical role in ILKAP-mediated tumor progression.ILKAP depletion also reduced extracellular acidification rates and altered glycolysis-related gene expression,highlighting its role in metabolic reprogramming.These findings suggest that ILKAP drives HCC malignancy by modulating PGAM1 and glycolysis,providing a potential therapeutic target for HCC treatment.Further elucidation of the ILKAP-PGAM1 axis may offer new strategies for liver cancer management.
文摘目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过表达组、miR-451a敲低组、温肺降浊方含药血清(WFJZF)组、miR-451a过表达+WFJZF组、miR-451a敲低+WFJZF组和miR-451a空载体组,双荧光素酶确定miR-451a和PGAM5的转录关系,免疫蛋白共沉淀确定PGAM5和Drp1的调控作用,慢病毒转染miR-451a于HT22细胞中,qRT-PCR确定转染效率,CCK-8测定缺糖缺氧状态下细胞活性的最佳时间点,qRT-PCR和Westem Blot检测HT22模型细胞内DGAM5-Drp1轴蛋白及mRNA的表达水平。结果:OGD/R诱导HT22细胞持续2 h为最佳时间点,miR-451a可以靶向调控PGAM5基因,PGAM5和Drp1具有相互调控作用。与正常对照组比较,OGD/R组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达上升,p-Drp1 Ser 616磷酸化表达上升(P<0.05),OPA1蛋白及mRNA表达下降,p-Drp1 Ser 637去磷酸化表达下降(P<0.05);与OGD/R组比较,miR-451a过表达+WFJZF组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达下降,p-Drp1 Ser 616磷酸化表达下降(P<0.05),OPA1蛋白及mRNA表达上升,p-Drp1 Ser 637去磷酸化表达上升(P<0.05)。结论:miR-451a可以靶向调控PGAM5-Drp1轴,miR-451a过表达+WFJZF组可以改善线粒体失衡状态,减少神经元过度损伤,发挥脑保护效应。
文摘目的:探究磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)变构抑制剂能否提高结肠癌细胞对奥沙利铂(oxaliplatin,OXA)的敏感性。方法:癌症药物敏感性基因组学(genomics of drug sensitivity in cancer,GDSC)和癌细胞系百科全书数据库(cancer cell line encyclopedia,CCLE)分析四种共识分子亚型(consensus molecular subgroups,CMS)结肠癌细胞对OXA敏感性和PGAM1的表达水平;采用CCK8法和Annexin V/PI双染法,探究PGAM1变构抑制剂HKB99对人源结肠癌细胞增殖和凋亡的作用;采用CCK8法分析HKB99联合OXA对结肠癌增殖的作用,并进行协同系数分析。结果:首先,GDSC和CCLE数据库分析结果显示,相对于CMS2和CMS3亚型,CMS1和CMS4亚型结肠癌细胞对OXA敏感的细胞比例降低,PGAM1表达升高,具有显著性差异(P<0.05)。进一步研究表明,HKB99显著抑制不同CMS类型的人源结肠癌SW480、SCUN2B和DLD-1细胞的增殖,其IC50均为1μM左右;Annexin V/PI双染法结果显示,与对照组相比,HKB99处理组DLD-1细胞的凋亡水平显著上升(P<0.01)。最后,CCK8法检测显示,与OXA单加药组相比,1.5μM的HKB99显著提高结肠癌DLD-1细胞对OXA的敏感性(IC_(50):3.26 vs0.37μM);协同系数分析结果显示,OXA在浓度1.56μM和25μM之间与HKB99的协同系数小于1。结论:本研究发现相对于CMS2和CMS3,CMS1和CMS4结肠癌细胞对OXA敏感性较低且PGAM1表达水平升高,PGAM1变构抑制剂HKB99增强结肠癌细胞对OXA的敏感性,提示HKB99有望提高结肠癌OXA化疗的敏感性,为患者个体化化疗提供新的治疗策略。
