Background:Intrahepatic cholangiocarcinoma(ICC)is a challenging can-cer with an increasing incidence.The Phase III TOPAZ-1/KEYNOTE-966study demonstrated chemo-immunotherapy(CIT)as a significant advance-ment,potentiall...Background:Intrahepatic cholangiocarcinoma(ICC)is a challenging can-cer with an increasing incidence.The Phase III TOPAZ-1/KEYNOTE-966study demonstrated chemo-immunotherapy(CIT)as a significant advance-ment,potentially replacing traditional chemotherapy for advanced biliary tractcancer.Ferroptosis is a crucial process that affects cancer cell survival andtherapy resistance.Although AKT hyperactivation is prevalent in numerouscancers,including ICC,its role in ferroptosis resistance remains unclear.Thisstudy explored whether targeting ferroptosis can enhance CIT response rates,specifically in ICC patients with AKT hyperactivation.Methods:In vivo metabolic CRISPR screening in a KrasG12D/Tp53−/−ICCmouse model was used to identify primary regulators of ferroptosis duringCIT(gemcitabine,cisplatin,and anti-mouse programmed cell death 1 ligand 1).Phosphoenolpyruvate carboxykinase 1(PCK1)was assessed for its role in fer-roptosis and treatment resistance in preclinical models under AKT activationlevels.Molecular and biochemical techniques were used to explore PCK1-relatedresistance mechanisms in AKT-hyperactivated ICC.Results:Under AKT hyperactivation condition,phosphorylated PCK1(pPCK1)promoted metabolic reprogramming,enhancing ubiquinol and menaquinone-4synthesisthrough the mevalonate(MVA)pathway.This cascade was mediated bythe pPCK1-pLDHA-SPRINGlac axis.Inhibiting PCK1 phosphorylation or usingsimvastatin significantly augmented CIT efficacy in preclinical models.Clini-cal data further indicated that phosphorylated AKT(pAKT)-pPCK1 levels mightserve as a biomarker to predict CIT response in ICC.Conclusion:Thisstudy identified the pAKT-pPCK1-pLDHA-SPRINGlac axis asa novel mechanism driving ferroptosis resistance in AKT-hyperactivated ICC byassociating glycolytic activation with MVA flux reprogramming.Targeting thisaxis,potentially through statin-based therapies,may offer a strategy to sensitizeICC cells to ferroptosis and improve treatment outcomes.展开更多
目的:体内体外联合探讨黄芪-丹参通过调控线粒体生物发生改善环孢素A(CsA)慢性肾毒性的作用。方法:体内将雄性C57BL/6小鼠随机分为5组:正常组、模型组、黄芪-丹参低剂量组、黄芪-丹参高剂量组及缬沙坦组。体外以黄芪-丹参干预CsA诱导的...目的:体内体外联合探讨黄芪-丹参通过调控线粒体生物发生改善环孢素A(CsA)慢性肾毒性的作用。方法:体内将雄性C57BL/6小鼠随机分为5组:正常组、模型组、黄芪-丹参低剂量组、黄芪-丹参高剂量组及缬沙坦组。体外以黄芪-丹参干预CsA诱导的人肾小管近端上皮细胞HK-2肾毒性模型,进一步分别在体内、体外敲减PGC1-α后分为PGC1-α敲减组、PGC1-α敲减空载体组、PGC1-α敲减+黄芪-丹参高剂量组、PGC1-α敲减空载体+黄芪-丹参高剂量组,观察黄芪-丹参调控PGC1-α/Pck1/TGF-β通路稳定线粒体生物发生的作用。结果:①体内研究发现,与模型组比较,黄芪-丹参高剂量组显著回调了BUN、Scr、UACR、NAG、IL-6及M D A(P<0.01,P<0.05),改善了肾小球基底膜增厚、肾小管萎缩、肾组织凋亡及胶原沉积。与PGC1-α敲减空载体组比较,PGC1-α敲减组Pck1蛋白表达下调,TGF-β1、Caspase9蛋白表达上调;与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参高剂量组显著改善了上述指标(P<0.05,P<0.01)。此外,与PGC1-α敲减空载体组比较,PGC1-α敲减组Smad7蛋白表达量减少,COL1A1、COL3A1、FN、Smad3蛋白表达量增多;与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参高剂量组上述指标的表达均得到改善。②体外研究发现,CsA诱导下HK-2凋亡增加、迁移增强、线粒体数量减少,结构损伤、线粒体膜电位下降,线粒体呼吸及ATP产生障碍。黄芪-丹参干预后均有不同程度的改善。与PGC1-α敲减空载体组比较,PGC1-α敲减组Pck1蛋白表达下调,TGF-β1、Caspase9蛋白表达上调,线粒体膜电位下降(P<0.05,P<0.01);与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参组显著改善了上述指标(P<0.01)。此外,与PGC1-α敲减空载体组比较,PGC1-α敲减组E-cadherin蛋白表达量减少,α-SMA蛋白表达量增多;与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参组上述指标的表达均得到改善。结论:黄芪-丹参能够通过调控PGC1-α/Pck1/TGF-β通路稳定线粒体生物发生,改善肾小管上皮细胞EMT,抑制ECM沉积,保护肾功能。展开更多
The last few decades have witnessed an advancement in our understanding of multiple cancer cell pathways related to metabolic reprogramming.One of the most important cancer hallmarks,including aerobic glycolysis(the W...The last few decades have witnessed an advancement in our understanding of multiple cancer cell pathways related to metabolic reprogramming.One of the most important cancer hallmarks,including aerobic glycolysis(the Warburg effect),the central carbon pathway,and multiple-branch metabolic pathway remodeling,enables tumor growth,progression,and metastasis.Phosphoenolpyruvate carboxykinase1(PCK1),a key rate-limiting enzyme in gluconeogenesis,catalyzes the conversion of oxaloacetate to phosphoenolpyruvate.PCK1 expression in gluconeogenic tissues is tightly regulated during fasting.In tumor cells,PCK1 is regulated in a cell-autonomous manner rather than by hormones or nutrients in the extracellular environment.Interestingly,PCK1has ananti-oncogenic role in gluconeogenic organs(the liver and kidneys),but a tumor-promoting role in cancers arising from non-gluconeogenic organs.Recent studies have revealed that PCK1 has metabolic and non-metabolic roles in multiple signaling networks linking metabolic and oncogenic pathways.Aberrant PCK1 expression results in the activation of oncogenic pathways,accompanied by metabolic reprogramming,to maintain tumorigenesis.In this review,we summarize the mechanisms underlying PCK1 expression and regulation,and clarify the crosstalk between aberrant PCK1 expression,metabolic rewiring,and signaling pathway activation.In addition,we highlight the clinical relevance of PCK1 and its value as a putative cancer therapeutic target.展开更多
Phosphoenolpyruvate carboxykinase 1(PCK1),a step limiting enzyme of gluconeogenesis,is downregulated in hepatocellular carcinoma(HCC).Overexpression of PCK1 has been shown to suppress hepatoma cell growth,but the unde...Phosphoenolpyruvate carboxykinase 1(PCK1),a step limiting enzyme of gluconeogenesis,is downregulated in hepatocellular carcinoma(HCC).Overexpression of PCK1 has been shown to suppress hepatoma cell growth,but the underlying mechanism remains unclear.We used recombinant adenovirus overexpressing PCK1 or GFP in Huh7 cells,and the differentially expressed genes(DEGs)were identified by RNA-Seq.180 were upregulated by PCK1 overexpression,whereas 316 were downregulated.Pathway analysis illustrated that PCK1 was closely correlated with Wnt signaling pathway and TGF-beta signaling pathway.Hence,Wnt signaling pathway and its downstream component,FZD2,FZD6,FZD7 and b-catenin were confirmed by qRT-PCR and Western blot.In vivo we also observed that PCK1 had restrained tumor growth as a result of decreasing expression of b-catenin.Whole-transcriptomic profile analysis discovered that overexpression of PCK1 downregulates several oncogenic signaling pathways in HCC,providing potential therapeutic targets for improving HCC therapy.展开更多
目的研究固醇调节元件结合蛋白1c(sterol regulatory element binding protein 1c,SREBP1c)对肠道糖异生的调控作用。方法饥饿处理C57BL/6小鼠、SREBP1c纯合敲除型(SREBP1c-KO)和同窝野生型(SREBP1c-WT)小鼠,通过qPCR、Western blot检...目的研究固醇调节元件结合蛋白1c(sterol regulatory element binding protein 1c,SREBP1c)对肠道糖异生的调控作用。方法饥饿处理C57BL/6小鼠、SREBP1c纯合敲除型(SREBP1c-KO)和同窝野生型(SREBP1c-WT)小鼠,通过qPCR、Western blot检测糖异生途径限速酶葡萄糖-6-磷酸酶(glucose-6-phosphatase catalytic,G6PC)和磷酸烯醇式丙酮酸羧激酶(phosphoenolpyruvate carboxykinase 1,PCK1)在肝脏、空肠和回肠组织中的表达水平;在肠上皮细胞CaCo-2中过表达或敲低SREBP1c,检测细胞内G6PC和PCK1表达水平。结果饥饿处理后,C57BL/6小鼠空肠、回肠组织中G6PC、PCK1和SREBP1c表达水平均显著增加(P<0.05)。SREBP1c基因敲除后,小鼠空肠、回肠组织中由饥饿诱导的糖异生限速酶G6PC和PCK1的表达明显下调(P<0.05)。在肠上皮细胞CaCo-2中过表达SREBP1c,可显著上调糖异生途径关键酶G6PC和PCK1的表达,促进细胞内葡萄糖的产生(P<0.05);反之,敲低SREBP1c的表达,可明显下调G6PC和PCK1,抑制细胞内葡萄糖的产生(P<0.05)。结论饥饿状态下,SREBP1c可调控肠上皮细胞中糖异生限速酶G6PC和PCK1的表达,进而影响葡萄糖的生成,表明SREBP1c可能参与肠道糖异生的调控,机制有待进一步探索。展开更多
Metabolic reprogramming plays a central role in tumors.However,the key drivers modulating reprogramming of gluconeogenesis/lipogenesis are poorly understood.Here,we try to identify the mechanism by which histone acety...Metabolic reprogramming plays a central role in tumors.However,the key drivers modulating reprogramming of gluconeogenesis/lipogenesis are poorly understood.Here,we try to identify the mechanism by which histone acetyltransferase 1(HAT1)confers reprogramming of gluconeogenesis/lipogenesis in liver cancer.Diethylnitrosamine(DEN)/carbon tetrachloride(CCl4)-induced hepatocarcinogenesis was hardly observed in HAT1-knockout mice.Multi-omics identified that HAT1 modulated gluconeogenesis and lipogenesis in liver.Protein phosphatase 2 scaffold subunit alpha(PPP2R1A)promoted gluconeogenesis and inhibited lipogenesis by phosphoenolpyruvate carboxykinase 1(PCK1)serine 90 dephosphorylation to suppress the tumor growth.HAT1 succinylated PPP2R1A at lysine 541(K541)to block the assembly of protein phosphatase 2A(PP2A)holoenzyme and interaction with PCK1,resulting in the depression of dephosphorylation of PCK1.HAT1-succinylated PPP2R1A contributed to the remodeling of gluconeogenesis/lipogenesis by PCK1 serine 90 phosphorylation,leading to the inhibition of gluconeogenic enzyme activity and activating sterol regulatory element-binding protein 1(SREBP1)nuclear accumulation-induced lipogenesis gene expression,which enhanced the tumor growth.In conclusion,succinylation of PPP2R1A lysine 541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling through PCK1 S90 phosphorylation to support liver cancer.Our finding provides new insights into the mechanism by which post-translational modifications(PTMs)confer the conversion of tumor suppressor function to oncogene.展开更多
基金National Natural Science Foundation of China,Grant/Award Numbers:81874149,82171834,82303832,82403618China Postdoctoral Science Foundation,Grant/Award Number:2024M761039+5 种基金China National Postdoctoral Program for Innovative Talents,Grant/Award Number:BX20240129Postdoctor Project of Hubei Province,Grant/Award Number:2024HBBHCXA035Knowledge Innovation Program of Wuhan-ShuguangProject,Grant/Award Number:2022020801020456Key Research and Development Program of Social Development of Jiangsu Province,Grant/Award Number:BE2022725Suzhou Science and Technology Development Plan Project,Grant/Award Number:SKY2023049Jiangsu Province Seventh 333 High Level(Second Level)Talents Project。
文摘Background:Intrahepatic cholangiocarcinoma(ICC)is a challenging can-cer with an increasing incidence.The Phase III TOPAZ-1/KEYNOTE-966study demonstrated chemo-immunotherapy(CIT)as a significant advance-ment,potentially replacing traditional chemotherapy for advanced biliary tractcancer.Ferroptosis is a crucial process that affects cancer cell survival andtherapy resistance.Although AKT hyperactivation is prevalent in numerouscancers,including ICC,its role in ferroptosis resistance remains unclear.Thisstudy explored whether targeting ferroptosis can enhance CIT response rates,specifically in ICC patients with AKT hyperactivation.Methods:In vivo metabolic CRISPR screening in a KrasG12D/Tp53−/−ICCmouse model was used to identify primary regulators of ferroptosis duringCIT(gemcitabine,cisplatin,and anti-mouse programmed cell death 1 ligand 1).Phosphoenolpyruvate carboxykinase 1(PCK1)was assessed for its role in fer-roptosis and treatment resistance in preclinical models under AKT activationlevels.Molecular and biochemical techniques were used to explore PCK1-relatedresistance mechanisms in AKT-hyperactivated ICC.Results:Under AKT hyperactivation condition,phosphorylated PCK1(pPCK1)promoted metabolic reprogramming,enhancing ubiquinol and menaquinone-4synthesisthrough the mevalonate(MVA)pathway.This cascade was mediated bythe pPCK1-pLDHA-SPRINGlac axis.Inhibiting PCK1 phosphorylation or usingsimvastatin significantly augmented CIT efficacy in preclinical models.Clini-cal data further indicated that phosphorylated AKT(pAKT)-pPCK1 levels mightserve as a biomarker to predict CIT response in ICC.Conclusion:Thisstudy identified the pAKT-pPCK1-pLDHA-SPRINGlac axis asa novel mechanism driving ferroptosis resistance in AKT-hyperactivated ICC byassociating glycolytic activation with MVA flux reprogramming.Targeting thisaxis,potentially through statin-based therapies,may offer a strategy to sensitizeICC cells to ferroptosis and improve treatment outcomes.
文摘目的:体内体外联合探讨黄芪-丹参通过调控线粒体生物发生改善环孢素A(CsA)慢性肾毒性的作用。方法:体内将雄性C57BL/6小鼠随机分为5组:正常组、模型组、黄芪-丹参低剂量组、黄芪-丹参高剂量组及缬沙坦组。体外以黄芪-丹参干预CsA诱导的人肾小管近端上皮细胞HK-2肾毒性模型,进一步分别在体内、体外敲减PGC1-α后分为PGC1-α敲减组、PGC1-α敲减空载体组、PGC1-α敲减+黄芪-丹参高剂量组、PGC1-α敲减空载体+黄芪-丹参高剂量组,观察黄芪-丹参调控PGC1-α/Pck1/TGF-β通路稳定线粒体生物发生的作用。结果:①体内研究发现,与模型组比较,黄芪-丹参高剂量组显著回调了BUN、Scr、UACR、NAG、IL-6及M D A(P<0.01,P<0.05),改善了肾小球基底膜增厚、肾小管萎缩、肾组织凋亡及胶原沉积。与PGC1-α敲减空载体组比较,PGC1-α敲减组Pck1蛋白表达下调,TGF-β1、Caspase9蛋白表达上调;与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参高剂量组显著改善了上述指标(P<0.05,P<0.01)。此外,与PGC1-α敲减空载体组比较,PGC1-α敲减组Smad7蛋白表达量减少,COL1A1、COL3A1、FN、Smad3蛋白表达量增多;与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参高剂量组上述指标的表达均得到改善。②体外研究发现,CsA诱导下HK-2凋亡增加、迁移增强、线粒体数量减少,结构损伤、线粒体膜电位下降,线粒体呼吸及ATP产生障碍。黄芪-丹参干预后均有不同程度的改善。与PGC1-α敲减空载体组比较,PGC1-α敲减组Pck1蛋白表达下调,TGF-β1、Caspase9蛋白表达上调,线粒体膜电位下降(P<0.05,P<0.01);与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参组显著改善了上述指标(P<0.01)。此外,与PGC1-α敲减空载体组比较,PGC1-α敲减组E-cadherin蛋白表达量减少,α-SMA蛋白表达量增多;与PGC1-α敲减组比较,PGC1-α敲减+黄芪-丹参组上述指标的表达均得到改善。结论:黄芪-丹参能够通过调控PGC1-α/Pck1/TGF-β通路稳定线粒体生物发生,改善肾小管上皮细胞EMT,抑制ECM沉积,保护肾功能。
基金This work was supported by The China National Natural Science Foundation,China(No.82073251,82072286,81872270)The Natural Science Foundation Project of Chongqing,China(No.cstc2019jscx-dxwtBX0019,cstc2019jcyj-msxmX0587)+2 种基金The Kuanren Talents Program of The Second Affiliated Hospital of Chongqing Medical University,ChinaThe Future Medical Youth Innovation Team of Chongqing Medical University,China(No.W0036,W0101)The Science and Technology Research Program of Chongqing Municipal Education Commission,China(No.HZ2021006,KJZD-M202000401,KJQN201900429).
文摘The last few decades have witnessed an advancement in our understanding of multiple cancer cell pathways related to metabolic reprogramming.One of the most important cancer hallmarks,including aerobic glycolysis(the Warburg effect),the central carbon pathway,and multiple-branch metabolic pathway remodeling,enables tumor growth,progression,and metastasis.Phosphoenolpyruvate carboxykinase1(PCK1),a key rate-limiting enzyme in gluconeogenesis,catalyzes the conversion of oxaloacetate to phosphoenolpyruvate.PCK1 expression in gluconeogenic tissues is tightly regulated during fasting.In tumor cells,PCK1 is regulated in a cell-autonomous manner rather than by hormones or nutrients in the extracellular environment.Interestingly,PCK1has ananti-oncogenic role in gluconeogenic organs(the liver and kidneys),but a tumor-promoting role in cancers arising from non-gluconeogenic organs.Recent studies have revealed that PCK1 has metabolic and non-metabolic roles in multiple signaling networks linking metabolic and oncogenic pathways.Aberrant PCK1 expression results in the activation of oncogenic pathways,accompanied by metabolic reprogramming,to maintain tumorigenesis.In this review,we summarize the mechanisms underlying PCK1 expression and regulation,and clarify the crosstalk between aberrant PCK1 expression,metabolic rewiring,and signaling pathway activation.In addition,we highlight the clinical relevance of PCK1 and its value as a putative cancer therapeutic target.
基金We would like to thank Dr.Tong-Chuan He(University of Chicago,USA)for providing the AdEasy system.This study was supported by research grants from China National Natural Science Foundation(grant nos.81602417 to KW,and 81872270 and 81572683 to NT),the Major National S&T program(2017ZX10202203-004 to NT)Natural Science Foundation Project of CQ CSTC(grant no.cstc2018jcyjAX0254 to NT)The Program for Innovation Team of Higher Education in Chongqing(grant no.CXTDX201601015).
文摘Phosphoenolpyruvate carboxykinase 1(PCK1),a step limiting enzyme of gluconeogenesis,is downregulated in hepatocellular carcinoma(HCC).Overexpression of PCK1 has been shown to suppress hepatoma cell growth,but the underlying mechanism remains unclear.We used recombinant adenovirus overexpressing PCK1 or GFP in Huh7 cells,and the differentially expressed genes(DEGs)were identified by RNA-Seq.180 were upregulated by PCK1 overexpression,whereas 316 were downregulated.Pathway analysis illustrated that PCK1 was closely correlated with Wnt signaling pathway and TGF-beta signaling pathway.Hence,Wnt signaling pathway and its downstream component,FZD2,FZD6,FZD7 and b-catenin were confirmed by qRT-PCR and Western blot.In vivo we also observed that PCK1 had restrained tumor growth as a result of decreasing expression of b-catenin.Whole-transcriptomic profile analysis discovered that overexpression of PCK1 downregulates several oncogenic signaling pathways in HCC,providing potential therapeutic targets for improving HCC therapy.
文摘目的研究固醇调节元件结合蛋白1c(sterol regulatory element binding protein 1c,SREBP1c)对肠道糖异生的调控作用。方法饥饿处理C57BL/6小鼠、SREBP1c纯合敲除型(SREBP1c-KO)和同窝野生型(SREBP1c-WT)小鼠,通过qPCR、Western blot检测糖异生途径限速酶葡萄糖-6-磷酸酶(glucose-6-phosphatase catalytic,G6PC)和磷酸烯醇式丙酮酸羧激酶(phosphoenolpyruvate carboxykinase 1,PCK1)在肝脏、空肠和回肠组织中的表达水平;在肠上皮细胞CaCo-2中过表达或敲低SREBP1c,检测细胞内G6PC和PCK1表达水平。结果饥饿处理后,C57BL/6小鼠空肠、回肠组织中G6PC、PCK1和SREBP1c表达水平均显著增加(P<0.05)。SREBP1c基因敲除后,小鼠空肠、回肠组织中由饥饿诱导的糖异生限速酶G6PC和PCK1的表达明显下调(P<0.05)。在肠上皮细胞CaCo-2中过表达SREBP1c,可显著上调糖异生途径关键酶G6PC和PCK1的表达,促进细胞内葡萄糖的产生(P<0.05);反之,敲低SREBP1c的表达,可明显下调G6PC和PCK1,抑制细胞内葡萄糖的产生(P<0.05)。结论饥饿状态下,SREBP1c可调控肠上皮细胞中糖异生限速酶G6PC和PCK1的表达,进而影响葡萄糖的生成,表明SREBP1c可能参与肠道糖异生的调控,机制有待进一步探索。
基金supported by the National Natural Science Foundation of China(Nos.82103066,82372818,82303210,and 82302887)Tianjin Key Medical Discipline(Specialty)Construction Project(TJYXZDXK-009A,China).
文摘Metabolic reprogramming plays a central role in tumors.However,the key drivers modulating reprogramming of gluconeogenesis/lipogenesis are poorly understood.Here,we try to identify the mechanism by which histone acetyltransferase 1(HAT1)confers reprogramming of gluconeogenesis/lipogenesis in liver cancer.Diethylnitrosamine(DEN)/carbon tetrachloride(CCl4)-induced hepatocarcinogenesis was hardly observed in HAT1-knockout mice.Multi-omics identified that HAT1 modulated gluconeogenesis and lipogenesis in liver.Protein phosphatase 2 scaffold subunit alpha(PPP2R1A)promoted gluconeogenesis and inhibited lipogenesis by phosphoenolpyruvate carboxykinase 1(PCK1)serine 90 dephosphorylation to suppress the tumor growth.HAT1 succinylated PPP2R1A at lysine 541(K541)to block the assembly of protein phosphatase 2A(PP2A)holoenzyme and interaction with PCK1,resulting in the depression of dephosphorylation of PCK1.HAT1-succinylated PPP2R1A contributed to the remodeling of gluconeogenesis/lipogenesis by PCK1 serine 90 phosphorylation,leading to the inhibition of gluconeogenic enzyme activity and activating sterol regulatory element-binding protein 1(SREBP1)nuclear accumulation-induced lipogenesis gene expression,which enhanced the tumor growth.In conclusion,succinylation of PPP2R1A lysine 541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling through PCK1 S90 phosphorylation to support liver cancer.Our finding provides new insights into the mechanism by which post-translational modifications(PTMs)confer the conversion of tumor suppressor function to oncogene.
