乙肝病毒X蛋白结合蛋白通过下调PEPCK的表达抑制肝脏糖异生（英文）
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摘要
乙肝病毒X蛋白结合蛋白(hepatitis B X-interacting protein,HBXIP)可以调节乳腺癌中糖代谢重编程.为了研究HBXIP在生理条件下对糖代谢的调节作用及机制,本研究利用Cre/lox P重组酶系统成功构建了肝脏组织中HBXIP特异敲除小鼠.当小鼠接受刺激后,与正常组小鼠相比,肝脏HBXIP敲除小鼠表现基础糖代谢功能异常,如葡萄糖、丙酮酸;相对于对照小鼠,肝脏HBXIP敲除小鼠对糖异生和胰岛素耐受性减弱. RT-PCR、Western blot实验和免疫组化实验结果表明,HBXIP敲除小鼠肝脏组织中糖异生关键酶磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxykinase,PEPCK)表达显著增加. QRT-PCR分析30例临床肝组织中HBXIP m RNA和PEPCK m RNA表达水平发现,HBXIP与PEPCK表达水平呈负相关.荧光素酶报告基因实验和Ch IP实验结果表明HBXIP可以在基因转录水平调节PEPCK表达.以上结果表明,HBXIP通过调节糖异生关键酶PEPCK的表达参与调控小鼠肝脏糖异生.
Hepatitis B X-interacting protein(HBXIP) is able to mediate glucose metabolism reprogramming in breast cancer. To investigate the physiological functions of HBXIP in regulation of glucose metabolism, we generated liver-specific HBXIP conditional knockout C57 BL/6 mice using Cre/lox P approach. Liver HBXIP-/-mice exhibited a phenotype of glycometabolic dysregulation, such as higher fasting blood glucose level, accumulation of hepatic glycogen, recession of blood glucose profiles, and elevation of gluconeogenesis. The expression levels of PEPCK were remarkably up-regulated in the liver tissues of HBXIP-/-mice.Then, we validated that HBXIP expressions were negatively correlated with those of PEPCK in 30 clinical liver tissues.Mechanistically, luciferase reporter gene assays and Ch IP assays showed that HBXIP could inhibit the expression PEPCK at transcription level. Taken together, our findings indicate that HBXIP restrains hepatic gluconeogenesis through suppressing the expression of PEPCK.
Hepatitis B X-interacting protein(HBXIP) is able to mediate glucose metabolism reprogramming in breast cancer. To investigate the physiological functions of HBXIP in regulation of glucose metabolism, we generated liver-specific HBXIP conditional knockout C57 BL/6 mice using Cre/lox P approach. Liver HBXIP-/-mice exhibited a phenotype of glycometabolic dysregulation, such as higher fasting blood glucose level, accumulation of hepatic glycogen, recession of blood glucose profiles, and elevation of gluconeogenesis. The expression levels of PEPCK were remarkably up-regulated in the liver tissues of HBXIP-/-mice.Then, we validated that HBXIP expressions were negatively correlated with those of PEPCK in 30 clinical liver tissues.Mechanistically, luciferase reporter gene assays and Ch IP assays showed that HBXIP could inhibit the expression PEPCK at transcription level. Taken together, our findings indicate that HBXIP restrains hepatic gluconeogenesis through suppressing the expression of PEPCK.
引文
[1]Cheng Y S,Seibert O,Kloting N,et al.PPP2R5C couples hepatic glucose and lipid homeostasis.Plos Genetics,2015,11(10):e1005561
[2]Calabuig-Navarro V,Yamauchi J,Lee S,et al.Forkhead Box O6(Fox O6)depletion attenuates hepatic gluconeogenesis and protects against fat-induced glucose disorder in mice.The Journal of Biological Chemistry,2015,290(25):15581-15594
[3]Ekberg K,Landau B R,Wajngot A,et al.Contributions by kidney and liver to glucose production in the postabsorptive state and after60 h of fasting.Diabetes,1999,48(2):292-298
[4]White M F.IRS proteins and the common path to diabetes.American Journal of Physiology Endocrinology and Metabolism,2002,283(3):E413-422
[5]Liu T Y,Shi C X,Gao R,et al.Irisin inhibits hepatic gluconeogenesis and increases glycogen synthesis via the PI3K/Akt pathway in type 2 diabetic mice and hepatocytes.Clinical Science,2015,129(10):839-850
[6]Fujii R,Zhu C,Wen Y,et al.HBXIP,cellular target of hepatitis Bvirus oncoprotein,is a regulator of centrosome dynamics and cytokinesis.Cancer Research,2006,66(18):9099-9107
[7]Melegari M,Scaglioni P P,Wands J R.Cloning and characterization of a novel hepatitis B virus x binding protein that inhibits viral replication.Journal of Virology,1998,72(3):1737-1743
[8]Marusawa H,Matsuzawa S,Welsh K,et al.HBXIP functions as a cofactor of survivin in apoptosis suppression.The EMBO Journal,2003,22(11):2729-2740
[9]Bar-Peled L,Schweitzer L D,Zoncu R,et al.Ragulator is a GEFfor the rag GTPases that signal amino acid levels to m TORC1.Cell,2012,150(6):1196-1208
[10]Liu F,Zhang W,You X,et al.The oncoprotein HBXIP promotes glucose metabolism reprogramming via downregulating SCO2and PDHA1 in breast cancer.Oncotarget,2015,6(29):27199-27213
[11]Nagy A.Cre recombinase:the universal reagent for genome tailoring.Genesis,2000,26(2):99-109
[12]Meng F,Shi L,Cheng X,et al.Surfactant protein A promoter directs the expression of Cre recombinase in brain microvascular endothelial cells of transgenic mice.Matrix Biology,2007,26(1):54-57
[13]Shi H,Fang R,Li Y,et al.The oncoprotein HBXIP suppresses gluconeogenesis through modulating PCK1 to enhance the growth of hepatoma cells.Cancer Letters,2016,382(2):147-156
[14]Shi H,Li Y,Feng G,et al.The oncoprotein HBXIP up-regulates FGF4 through activating transcriptional factor Sp1 to promote the migration of breast cancer cells.Biochemical and Biophysical Research Communications,2016,471(1):89-94
[15]Li L,Fang R,Liu B,et al.Deacetylation of tumor-suppressor MST1 in Hippo pathway induces its degradation through HBXIP-elevated HDAC6 in promotion of breast cancer growth.Oncogene,2016,35(31):4048-4057
[16]Li Y,Wang Z,Shi H,et al.HBXIP and LSD1 scaffolded by lnc RNA hotair mediate transcriptional activation by c-Myc.Cancer Research,2016,76(2):293-304
[17]Wang T,Geng S L,Guan Y M,et al.Deacetylation of metabolic enzymes by Sirt2 modulates pyruvate homeostasis to extend insect lifespan.Aging,2018,10(5):1053-1072
[18]Li K,Zhang J,Yu J,et al.Micro RNA-214 suppresses gluconeogenesis by targeting activating transcriptional factor 4.The Journal of Biological Chemistry,2015,290(13):8185-8195
[19]Liu Q,Lu W,Yang C,et al.HBXIP activates the PPARdelta/NF-kappa B feedback loop resulting in cell proliferation.Oncotarget,2018,9(1):404-417
[20]Liu B W,Wang T J,Li L L,et al.Oncoprotein HBXIP induces PKM2 via transcription factor E2F1 to promote cell proliferation in ER-positive breast cancer.Acta Pharmacologica Sinica,2018,doi:10.1038/s41401-018-0015-9
[21]Anyamaneeratch K,Rojvirat P,Sukjoi W,et al.Insights into transcriptional regulation of hepatic glucose production.International Review of Cell and Molecular Biology,2015,318:203-253
[22]Nordlie R C,Foster J D,Lange A J.Regulation of glucose production by the liver.Annual Review of Nutrition,1999,19:379-406
[23]Molero C,Valverde A M,Benito M,et al.Phosphoenolpyruvate carboxykinase and glucose-6-phosphate dehydrogenase expression in fetal hepatocyte primary cultures under proliferative conditions.Experimental Cell Research,1992,200(2):295-300
[24]Li H,Wang Z,Li Y,et al.Hepatitis B X-interacting protein promotes the formation of the insulin gene-transcribing protein complex Pdx-1/Neurod1 in animal pancreatic beta-cells.The Journal of Biological Chemistry,2018,293(6):2053-2065
[25]Wang Y,Fang R,Cui M,et al.The oncoprotein HBXIP upregulates YAP through activation of transcription factor c-Myb to promote growth of liver cancer.Cancer Letters,2017,385:234-242
[2]Calabuig-Navarro V,Yamauchi J,Lee S,et al.Forkhead Box O6(Fox O6)depletion attenuates hepatic gluconeogenesis and protects against fat-induced glucose disorder in mice.The Journal of Biological Chemistry,2015,290(25):15581-15594
[3]Ekberg K,Landau B R,Wajngot A,et al.Contributions by kidney and liver to glucose production in the postabsorptive state and after60 h of fasting.Diabetes,1999,48(2):292-298
[4]White M F.IRS proteins and the common path to diabetes.American Journal of Physiology Endocrinology and Metabolism,2002,283(3):E413-422
[5]Liu T Y,Shi C X,Gao R,et al.Irisin inhibits hepatic gluconeogenesis and increases glycogen synthesis via the PI3K/Akt pathway in type 2 diabetic mice and hepatocytes.Clinical Science,2015,129(10):839-850
[6]Fujii R,Zhu C,Wen Y,et al.HBXIP,cellular target of hepatitis Bvirus oncoprotein,is a regulator of centrosome dynamics and cytokinesis.Cancer Research,2006,66(18):9099-9107
[7]Melegari M,Scaglioni P P,Wands J R.Cloning and characterization of a novel hepatitis B virus x binding protein that inhibits viral replication.Journal of Virology,1998,72(3):1737-1743
[8]Marusawa H,Matsuzawa S,Welsh K,et al.HBXIP functions as a cofactor of survivin in apoptosis suppression.The EMBO Journal,2003,22(11):2729-2740
[9]Bar-Peled L,Schweitzer L D,Zoncu R,et al.Ragulator is a GEFfor the rag GTPases that signal amino acid levels to m TORC1.Cell,2012,150(6):1196-1208
[10]Liu F,Zhang W,You X,et al.The oncoprotein HBXIP promotes glucose metabolism reprogramming via downregulating SCO2and PDHA1 in breast cancer.Oncotarget,2015,6(29):27199-27213
[11]Nagy A.Cre recombinase:the universal reagent for genome tailoring.Genesis,2000,26(2):99-109
[12]Meng F,Shi L,Cheng X,et al.Surfactant protein A promoter directs the expression of Cre recombinase in brain microvascular endothelial cells of transgenic mice.Matrix Biology,2007,26(1):54-57
[13]Shi H,Fang R,Li Y,et al.The oncoprotein HBXIP suppresses gluconeogenesis through modulating PCK1 to enhance the growth of hepatoma cells.Cancer Letters,2016,382(2):147-156
[14]Shi H,Li Y,Feng G,et al.The oncoprotein HBXIP up-regulates FGF4 through activating transcriptional factor Sp1 to promote the migration of breast cancer cells.Biochemical and Biophysical Research Communications,2016,471(1):89-94
[15]Li L,Fang R,Liu B,et al.Deacetylation of tumor-suppressor MST1 in Hippo pathway induces its degradation through HBXIP-elevated HDAC6 in promotion of breast cancer growth.Oncogene,2016,35(31):4048-4057
[16]Li Y,Wang Z,Shi H,et al.HBXIP and LSD1 scaffolded by lnc RNA hotair mediate transcriptional activation by c-Myc.Cancer Research,2016,76(2):293-304
[17]Wang T,Geng S L,Guan Y M,et al.Deacetylation of metabolic enzymes by Sirt2 modulates pyruvate homeostasis to extend insect lifespan.Aging,2018,10(5):1053-1072
[18]Li K,Zhang J,Yu J,et al.Micro RNA-214 suppresses gluconeogenesis by targeting activating transcriptional factor 4.The Journal of Biological Chemistry,2015,290(13):8185-8195
[19]Liu Q,Lu W,Yang C,et al.HBXIP activates the PPARdelta/NF-kappa B feedback loop resulting in cell proliferation.Oncotarget,2018,9(1):404-417
[20]Liu B W,Wang T J,Li L L,et al.Oncoprotein HBXIP induces PKM2 via transcription factor E2F1 to promote cell proliferation in ER-positive breast cancer.Acta Pharmacologica Sinica,2018,doi:10.1038/s41401-018-0015-9
[21]Anyamaneeratch K,Rojvirat P,Sukjoi W,et al.Insights into transcriptional regulation of hepatic glucose production.International Review of Cell and Molecular Biology,2015,318:203-253
[22]Nordlie R C,Foster J D,Lange A J.Regulation of glucose production by the liver.Annual Review of Nutrition,1999,19:379-406
[23]Molero C,Valverde A M,Benito M,et al.Phosphoenolpyruvate carboxykinase and glucose-6-phosphate dehydrogenase expression in fetal hepatocyte primary cultures under proliferative conditions.Experimental Cell Research,1992,200(2):295-300
[24]Li H,Wang Z,Li Y,et al.Hepatitis B X-interacting protein promotes the formation of the insulin gene-transcribing protein complex Pdx-1/Neurod1 in animal pancreatic beta-cells.The Journal of Biological Chemistry,2018,293(6):2053-2065
[25]Wang Y,Fang R,Cui M,et al.The oncoprotein HBXIP upregulates YAP through activation of transcription factor c-Myb to promote growth of liver cancer.Cancer Letters,2017,385:234-242