在人肝癌 HepG2 细胞中稳定表达和功能表征 Na+-牛磺胆酸钠共转运的绿色荧光蛋白。
Stable expression and functional characterization of a Na+-taurocholate cotransporting green fluorescent protein in human hepatoblastoma HepG2 cells.
机构信息
Divisions of Clinical Pharmacology/Toxicology, Department of Medicine, University Hospital, CH-8091, Zürich, Switzerland,
出版信息
Cytotechnology. 2000 Oct;34(1-2):1-9. doi: 10.1023/A:1008152729133.
Abstract
Sodium-dependent uptake of bile acids from blood is aliver-specific function which is mediated by theNa(+)-taurocholate cotransporting polypeptide(Ntcp). We report the stable expression of aNa(+)-taurocholate cotransporting green fluorescentfusion protein in the human hepatoblastoma cell lineHepG2, normally lacking Ntcp expression. Ntcp-EGFPassociated green fluorescence colocalized with Ntcpimmunofluorescence in the plasma membrane. Intransfected HepG2 cells, the fusion protein mediatedthe sodium-dependent uptake of the bile acidtaurocholate (K(m): 24.6 mumol/l) and of the anionicsteroids estrone-3-sulfate and dehydroepiandrosteronesulfate. We conclude that the Ntcp-EGFP fusion proteinfollows the sorting route of Ntcp, is functionallyidentical to Ntcp and could be used to monitor proteintrafficking in living HepG2 cells.
摘要
胆酸从血液中的钠依赖性摄取是肝脏特异性功能，由 Na(+)-牛磺胆酸钠共转运蛋白 (Ntcp) 介导。我们报告了人肝癌细胞系 HepG2 中稳定表达的 Na(+)-牛磺胆酸钠共转运绿色荧光融合蛋白，该细胞系通常缺乏 Ntcp 表达。Ntcp-EGFP 相关的绿色荧光与质膜中的 Ntcp 免疫荧光共定位。在转染的 HepG2 细胞中，融合蛋白介导了胆汁酸牛磺胆酸钠（K(m)：24.6 μmol/l）和阴离子甾体雌酮-3-硫酸盐和脱氢表雄酮硫酸盐的钠依赖性摄取。我们得出结论，Ntcp-EGFP 融合蛋白遵循 Ntcp 的分拣途径，与 Ntcp 在功能上相同，可用于监测活 HepG2 细胞中的蛋白质运输。
相似文献
1
Stable expression and functional characterization of a Na+-taurocholate cotransporting green fluorescent protein in human hepatoblastoma HepG2 cells.在人肝癌 HepG2 细胞中稳定表达和功能表征 Na+-牛磺胆酸钠共转运的绿色荧光蛋白。
Cytotechnology. 2000 Oct;34(1-2):1-9. doi: 10.1023/A:1008152729133.
2
Protein kinase C induces endocytosis of the sodium taurocholate cotransporting polypeptide.蛋白激酶 C 诱导牛磺胆酸钠共转运多肽内吞。
Am J Physiol Gastrointest Liver Physiol. 2010 Aug;299(2):G320-8. doi: 10.1152/ajpgi.00180.2010. Epub 2010 Jun 10.
3
Molecular and functional characterization of bile acid transport in human hepatoblastoma HepG2 cells.人肝癌细胞系HepG2中胆汁酸转运的分子与功能特征
Hepatology. 1996 May;23(5):1053-60. doi: 10.1002/hep.510230518.
4
Short-term feedback regulation of bile salt uptake by bile salts in rodent liver.啮齿类动物肝脏中胆汁盐对胆汁盐摄取的短期反馈调节。
Hepatology. 2012 Dec;56(6):2387-97. doi: 10.1002/hep.25955.
5
Calnexin Depletion by Endoplasmic Reticulum Stress During Cholestasis Inhibits the Na-Taurocholate Cotransporting Polypeptide.胆汁淤积期间内质网应激导致钙连接蛋白耗竭，抑制牛磺胆酸钠共转运多肽。
Hepatol Commun. 2018 Oct 23;2(12):1550-1566. doi: 10.1002/hep4.1262. eCollection 2018 Dec.
6
In situ localization of the hepatocytic Na+/Taurocholate cotransporting polypeptide in rat liver.大鼠肝脏中肝细胞钠/牛磺胆酸盐共转运多肽的原位定位
Gastroenterology. 1994 Dec;107(6):1781-7. doi: 10.1016/0016-5085(94)90821-4.
7
Differential inhibition of rat and human Na+-dependent taurocholate cotransporting polypeptide (NTCP/SLC10A1)by bosentan: a mechanism for species differences in hepatotoxicity.波生坦对大鼠和人类钠依赖性牛磺胆酸共转运多肽（NTCP/SLC10A1）的差异性抑制：肝毒性物种差异的一种机制
J Pharmacol Exp Ther. 2007 Jun;321(3):1170-8. doi: 10.1124/jpet.106.119073. Epub 2007 Mar 20.
8
The Lipid Raft Component Stomatin Interacts with the Na Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake.脂质筏成分质膜蛋白 22(stomatin)与牛磺胆酸钠共转运多肽(NTCP)相互作用并调节胆汁盐摄取。
Cells. 2020 Apr 16;9(4):986. doi: 10.3390/cells9040986.
9
Kinetics of the bile acid transporter and hepatitis B virus receptor Na+/taurocholate cotransporting polypeptide (NTCP) in hepatocytes.肝细胞中胆汁酸转运蛋白和乙型肝炎病毒受体钠离子/牛磺胆酸钠共转运多肽（NTCP）的动力学。
J Hepatol. 2014 Oct;61(4):867-75. doi: 10.1016/j.jhep.2014.05.018. Epub 2014 May 15.
10
Hepatic uptake of conjugated bile acids is mediated by both sodium taurocholate cotransporting polypeptide and organic anion transporting polypeptides and modulated by intestinal sensing of plasma bile acid levels in mice.在小鼠中，结合型胆汁酸的肝脏摄取由牛磺胆酸钠共转运多肽和有机阴离子转运多肽介导，并受血浆胆汁酸水平的肠道感知调节。
Hepatology. 2017 Nov;66(5):1631-1643. doi: 10.1002/hep.29251. Epub 2017 Sep 29.
引用本文的文献
1
Detection of hepatitis B virus DNA and HBsAg from postmortem blood and bloodstains.从死后血液和血迹中检测乙型肝炎病毒DNA和乙肝表面抗原
Arch Virol. 2018 Mar;163(3):633-637. doi: 10.1007/s00705-017-3665-x. Epub 2017 Dec 2.
2
Hormesis in Cholestatic Liver Disease; Preconditioning with Low Bile Acid Concentrations Protects against Bile Acid-Induced Toxicity.胆汁淤积性肝病中的毒物兴奋效应；低胆汁酸浓度预处理可预防胆汁酸诱导的毒性作用。
PLoS One. 2016 Mar 7;11(3):e0149782. doi: 10.1371/journal.pone.0149782. eCollection 2016.
3
Human FXR regulates SHP expression through direct binding to an LRH-1 binding site, independent of an IR-1 and LRH-1.人 FXR 通过直接结合 LRH-1 结合位点而不是通过 IR-1 和 LRH-1 调节 SHP 的表达。
PLoS One. 2014 Feb 3;9(2):e88011. doi: 10.1371/journal.pone.0088011. eCollection 2014.
4
Pertussis toxin, an inhibitor of G(αi) PCR, inhibits bile acid- and cytokine-induced apoptosis in primary rat hepatocytes.百日咳毒素，一种 G(αi) 蛋白偶联受体的抑制剂，能够抑制原代大鼠肝细胞中胆汁酸和细胞因子诱导的细胞凋亡。
PLoS One. 2012;7(8):e43156. doi: 10.1371/journal.pone.0043156. Epub 2012 Aug 10.
5
In vitro investigation of the hepatobiliary disposition mechanisms of the antifungal agent micafungin in humans and rats.在体研究抗真菌药物米卡芬净在人体和大鼠中的肝胆处置机制。
Drug Metab Dispos. 2010 Oct;38(10):1848-56. doi: 10.1124/dmd.110.033811. Epub 2010 Jul 6.
6
Bile acids decrease intracellular bilirubin levels in the cholestatic liver: implications for bile acid-mediated oxidative stress.胆汁酸可降低胆汁淤积性肝脏中的细胞内胆红素水平：提示胆汁酸介导的氧化应激。
J Cell Mol Med. 2011 May;15(5):1156-65. doi: 10.1111/j.1582-4934.2010.01098.x. Epub 2010 May 28.
7
Inhibition of Na+-taurocholate Co-transporting polypeptide-mediated bile acid transport by cholestatic sulfated progesterone metabolites.胆汁酸转运蛋白介导的胆汁酸转运被胆淤积性硫酸化孕烯醇酮代谢物所抑制。
J Biol Chem. 2010 May 28;285(22):16504-12. doi: 10.1074/jbc.M109.072140. Epub 2010 Feb 20.
8
The solute carrier family SLC10: more than a family of bile acid transporters regarding function and phylogenetic relationships.溶质载体家族SLC10：就功能和系统发育关系而言，不止是一个胆汁酸转运体家族。
Naunyn Schmiedebergs Arch Pharmacol. 2006 Mar;372(6):413-31. doi: 10.1007/s00210-006-0043-8. Epub 2006 Mar 16.
9
The complexities of hepatic drug transport: current knowledge and emerging concepts.肝脏药物转运的复杂性：当前认知与新出现的概念
Pharm Res. 2004 May;21(5):719-35. doi: 10.1023/b:pham.0000026420.79421.8f.
10
Bile acid-activated nuclear receptor FXR suppresses apolipoprotein A-I transcription via a negative FXR response element.胆汁酸激活的核受体FXR通过负性FXR反应元件抑制载脂蛋白A-I的转录。
J Clin Invest. 2002 Apr;109(7):961-71. doi: 10.1172/JCI14505.
本文引用的文献
1
Hepatic transport of bile salts.胆汁盐的肝脏转运
Semin Liver Dis. 2000;20(3):273-92. doi: 10.1055/s-2000-9426.
2
Short-term regulation of bile acid uptake by microfilament-dependent translocation of rat ntcp to the plasma membrane.大鼠钠-牛磺胆酸共转运多肽（NTCP）通过微丝依赖的转位作用向质膜的转运对胆汁酸摄取的短期调节。
Hepatology. 1999 Jul;30(1):223-9. doi: 10.1002/hep.510300136.
3
Identification of a novel gene family encoding human liver-specific organic anion transporter LST-1.鉴定一个编码人肝脏特异性有机阴离子转运体LST-1的新基因家族。
J Biol Chem. 1999 Jun 11;274(24):17159-63. doi: 10.1074/jbc.274.24.17159.
4
Molecular cloning and functional characterization of two alternatively spliced Ntcp isoforms from mouse liver1.从小鼠肝脏中克隆并鉴定两种可变剪接的Ntcp异构体的分子及功能特性1。
Biochim Biophys Acta. 1999 Apr 14;1445(1):154-9. doi: 10.1016/s0167-4781(99)00029-9.
5
Lack of zonal uptake of estrone sulfate in enriched periportal and perivenous isolated rat hepatocytes.在富集的门静脉周围和肝静脉周围分离的大鼠肝细胞中硫酸雌酮缺乏区域摄取。
Drug Metab Dispos. 1999 Mar;27(3):336-41.
6
A GABAA receptor alpha1 subunit tagged with green fluorescent protein requires a beta subunit for functional surface expression.一个标记有绿色荧光蛋白的GABAA受体α1亚基需要一个β亚基来实现功能性表面表达。
J Biol Chem. 1998 Oct 30;273(44):28906-11. doi: 10.1074/jbc.273.44.28906.
7
Dehydroepiandrosterone sulfate (DHEAS): identification of a carrier protein in human liver and brain.硫酸脱氢表雄酮（DHEAS）：人肝脏和大脑中一种载体蛋白的鉴定。
FEBS Lett. 1998 Mar 13;424(3):173-6. doi: 10.1016/s0014-5793(98)00168-9.
8
Substrate specificity of sinusoidal bile acid and organic anion uptake systems in rat and human liver.大鼠和人肝脏中窦状隙胆汁酸及有机阴离子摄取系统的底物特异性
Hepatology. 1997 Dec;26(6):1667-77. doi: 10.1002/hep.510260641.
9
Identification and functional characterization of the promoter region of the human organic anion transporting polypeptide gene.人类有机阴离子转运多肽基因启动子区域的鉴定及功能特性分析
Hepatology. 1997 Oct;26(4):991-7. doi: 10.1002/hep.510260429.
10
ER-to-Golgi transport visualized in living cells.活细胞中内质网到高尔基体的转运可视化
Nature. 1997 Sep 4;389(6646):81-5. doi: 10.1038/38001.
Zotero 插件