Expression profile and down-regulation of argininosuccinate synthetase in hepatocellular carcinoma in a transgenic mouse model

Journal of Biomedical Science

Tsung-Sheng Su⁶  Chiu-Jung Huang³  Kong Bung Choo⁴  Alice Chien Chang²  Ting-Fen Tsai⁶  Miao-Zeng Huang⁵  Shih-Chang Shiue¹ 

[1] Institute of Microbiology & Immunology, National Yang-Ming University, Taipei, Taiwan;Institute of Neuroscience, National Yang-Ming University, Taipei, Taiwan;Graduate Institute of Biotechnology, Chinese Culture University, Taipei, Taiwan;Centre for Stem Cell Research, Universiti Tunku Abdul Rahman, Selangor, Malaysia;Department of Medical Research, Taipei Veterans General Hospital, Taipei 112, Taiwan;Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Taipei, Taiwan

关键词: GFP reporter gene;    Bacterial artificial chromosome;    Post-transcriptional regulation;    Subventricular zone;    Ventricular zone;    Brain expression map;    Embryo expression map;    Hepatocellular carcinoma;    Transgenic mouse model;    Argininosuccinate synthetase;   

Others  :  1133564
DOI  :  10.1186/s12929-015-0114-6

 received in 2014-10-27, accepted in 2015-01-12,  发布年份 2015

【 摘 要 】

Background

Argininosuccinate synthetase (ASS) participates in urea and nitric oxide production and is a rate-limiting enzyme in arginine biosynthesis. Regulation of ASS expression appears complex and dynamic. In addition to transcriptional regulation, a novel post-transcriptional regulation affecting nuclear precursor RNA stability has been reported. Moreover, many cancers, including hepatocellular carcinoma (HCC), have been found not to express ASS mRNA; therefore, they are auxotrophic for arginine. To study when and where ASS is expressed and whether post-transcriptional regulation is undermined in particular temporal and spatial expression and in pathological events such as HCC, we set up a transgenic mouse system with modified BAC (bacterial artificial chromosome) carrying the human ASS gene tagged with an EGFP reporter.

Results

We established and characterized the transgenic mouse models based on the use of two BAC-based EGFP reporter cassettes: a transcription reporter and a transcription/post-transcription coupled reporter. Using such a transgenic mouse system, EGFP fluorescence pattern in E14.5 embryo was examined. Profiles of fluorescence and that of Ass RNA in in situ hybridization were found to be in good agreement in general, yet our system has the advantages of sensitivity and direct fluorescence visualization. By comparing expression patterns between mice carrying the transcription reporter and those carrying the transcription/post-transcription couple reporter, a post-transcriptional up-regulation of ASS was found around the ventricular zone/subventricular zone of E14.5 embryonic brain. In the EGFP fluorescence pattern and mRNA level in adult tissues, tissue-specific regulation was found to be mainly controlled at transcriptional initiation. Furthermore, strong EGFP expression was found in brain regions of olfactory bulb, septum, habenular nucleus and choroid plexus of the young transgenic mice. On the other hand, in crossing to hepatitis B virus X protein (HBx)-transgenic mice, the Tg (ASS-EGFP, HBx) double transgenic mice developed HCC in which ASS expression was down-regulated, as in clinical samples.

Conclusions

The BAC transgenic mouse model described is a valuable tool for studying ASS gene expression. Moreover, this mouse model is a close reproduction of clinical behavior of ASS in HCC and is useful in testing arginine-depleting agents and for studies of the role of ASS in tumorigenesis.

【 授权许可】

   
2015 Shiue et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Morris SM Jr: Arginine: beyond protein. Am J Clin Nutr 2006, 83:508S-512S.
• [2]Husson A, Brasse-Lagnel C, Fairand A, Renouf S, Lavoinne A: Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycle. Eur J Biochem 2003, 270:1887-1899.
• [3]Haines RJ, Pendleton LC, Eichler DC: Argininosuccinate synthase: at the center of arginine metabolism. Int J Biochem Mol Biol 2011, 2:8-23.
• [4]Guei TR, Liu MC, Yang CP, Su TS: Identification of a liver-specific cAMP response element in the human argininosuccinate synthetase gene. Biochem Biophys Res Commun 2008, 377:257-261.
• [5]Brasse-Lagnel C, Lavoinne A, Loeber D, Fairand A, Bole-Feysot C, Deniel N, et al.: Glutamine and interleukin-1beta interact at the level of Sp1 and nuclear factor-kappaB to regulate argininosuccinate synthetase gene expression. FEBS J 2007, 274:5250-5262.
• [6]Tsai WB, Aiba I, Lee SY, Feun L, Savaraj N, Kuo MT: Resistance to arginine deiminase treatment in melanoma cells is associated with induced argininosuccinate synthetase expression involving c-Myc/HIF-1alpha/Sp4. Mol Cancer Ther 2009, 8:3223-3233.
• [7]Su TS, Beaudet AL, O’Brien WE: Increased translatable messenger ribonucleic acid for argininosuccinate synthetase in canavanine-resistant human cells. Biochemistry 1981, 20:2956-2960.
• [8]Tsai TF, Su TS: A nuclear post-transcriptional event responsible for overproduction of argininosuccinate synthetase in a canavanine-resistant variant of a human epithelial cell line. Eur J Biochem 1995, 229:233-238.
• [9]Tseng SH, Cheng CY, Huang MZ, Chung MY, Su TS: Modulation of formation of the 3′-end of the human argininosuccinate synthetase mRNA by GT-repeat polymorphism. Int J Biochem Mol Biol 2013, 4:179-190.
• [10]Lavoinne A, Meisse D, Quillard M, Husson A, Renouf S, Yassad A: Glutamine and regulation of gene expression in rat hepatocytes: the role of cell swelling. Biochimie 1998, 80:807-811.
• [11]Schimke RT: Enzymes of arginine metabolism in mammalian cell culture. i. repression of argininosuccinate synthetase and argininosuccinase. J Biol Chem 1964, 239:136-145.
• [12]Su TS, Bock HG, O’Brien WE, Beaudet AL: Cloning of cDNA for argininosuccinate synthetase mRNA and study of enzyme overproduction in a human cell line. J Biol Chem 1981, 256:11826-11831.
• [13]Dillon BJ, Prieto VG, Curley SA, Ensor CM, Holtsberg FW, Bomalaski JS, et al.: Incidence and distribution of argininosuccinate synthetase deficiency in human cancers: a method for identifying cancers sensitive to arginine deprivation. Cancer 2004, 100:826-833.
• [14]Delage B, Fennell DA, Nicholson L, McNeish I, Lemoine NR, Crook T, et al.: Arginine deprivation and argininosuccinate synthetase expression in the treatment of cancer. Int J Cancer 2010, 126:2762-2772.
• [15]Phillips MM, Sheaff MT, Szlosarek PW: Targeting arginine-dependent cancers with arginine-degrading enzymes: opportunities and challenges. Cancer Res Treat 2013, 45:251-262.
• [16]Yang H, Lin M, Xiong FX, Yang Y, Nie X, Zhou RL: Reduced expression of ASS is closely related to clinicopathological features and post-resectional survival of hepatocellular carcinoma. Oncol Lett 2010, 1:31-36.
• [17]Wu L, Li L, Meng S, Qi R, Mao Z, Lin M: Expression of argininosuccinate synthetase in patients with hepatocellular carcinoma. J Gastroenterol Hepatol 2013, 28:365-368.
• [18]Szlosarek PW, Klabatsa A, Pallaska A, Sheaff M, Smith P, Crook T, et al.: In vivo loss of expression of argininosuccinate synthetase in malignant pleural mesothelioma is a biomarker for susceptibility to arginine depletion. Clin Cancer Res 2006, 12:7126-7131.
• [19]Szlosarek PW, Grimshaw MJ, Wilbanks GD, Hagemann T, Wilson JL, Burke F, et al.: Aberrant regulation of argininosuccinate synthetase by TNF-alpha in human epithelial ovarian cancer. Int J Cancer 2007, 121:6-11.
• [20]Shiue SC, Huang MZ, Su TS: A transgenic approach to study argininosuccinate synthetase gene expression. J Biomed Sci 2014, 21:42. BioMed Central Full Text
• [21]Gust B, Challis GL, Fowler K, Kieser T, Chater KF: PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proc Natl Acad Sci U S A 2003, 100:1541-1546.
• [22]Wu BK, Li CC, Chen HJ, Chang JL, Jeng KS, Chou CK, et al.: Blocking of G1/S transition and cell death in the regenerating liver of Hepatitis B virus X protein transgenic mice. Biochem Biophys Res Commun 2006, 340:916-928.
• [23]Gong S, Zheng C, Doughty ML, Losos K, Didkovsky N, Schambra UB, et al.: A gene expression atlas of the central nervous system based on bacterial artificial chromosomes. Nature 2003, 425:917-925.
• [24]Ko MS, Kitchen JR, Wang X, Threat TA, Wang X, Hasegawa A, et al.: Large-scale cDNA analysis reveals phased gene expression patterns during preimplantation mouse development. Development 2000, 127:1737-1749.
• [25]Mouse Genome Informatics [http://www.informatics.jax.org/searches/probe.cgi?824472]
• [26]GenePaint.org [http://www.genepaint.org/Midline.htm]
• [27]Kaufman MH: The Atlas of Mouse Development. Academic, San Diego; 1992.
• [28]The Mouse Gene Expression Database (GXD) [http://www.informatics.jax.org/gxd/marker/MGI:88090]
• [29]Noctor SC, Martínez-Cerdeño V, Ivic L, Kriegstein AR: Cortical neurons arise in symmetric and asymmetric division zones and migrate through specific phases. Nat Neurosci 2004, 7:136-144.
• [30]Hennecke M, Kwissa M, Metzger K, Oumard A, Kröger A, Schirmbeck R, et al.: Composition and arrangement of genes define the strength of IRES-driven translation in bicistronic mRNAs. Nucleic Acids Res 2001, 29:3327-3334.
• [31]Shackleford JM, Schneyer CA: Structural and functional aspects of rodent salivary glands including two desert species. Am J Anat 1964, 115:279-307.
• [32]Maekawa M, Kamimura K, Nagano T: Peritubular myoid cells in the testis: their structure and function. Arch Histol Cytol 1996, 59:1-13.
• [33]Haider SG: Cell biology of Leydig cells in the testis. Int Rev Cytol 2004, 233:181-241.
• [34]Nagasaki A, Gotoh T, Takeya M, Yu Y, Takiguchi M, Matsuzaki H, et al.: Coinduction of nitric oxide synthase, argininosuccinate synthetase, and argininosuccinate lyase in lipopolysaccharide-treated rats. RNA blot, immunoblot, and immunohistochemical analyses. J Biol Chem 1996, 271:2658-2662.
• [35]Nakamura H, Saheki T, Ichiki H, Nakata K, Nakagawa S: Immunocytochemical localization of argininosuccinate synthetase in the rat brain. J Comp Neurol 1991, 312:652-679.
• [36]Heneka MT, Wiesinger H, Dumitrescu-Ozimek L, Riederer P, Feinstein DL, Klockgether T: Neuronal and glial coexpression of argininosuccinate synthetase and inducible nitric oxide synthase in Alzheimer disease. J Neuropathol Exp Neurol 2001, 60:906-916.
• [37]Heneka MT, Schmidlin A, Wiesinger H: Induction of argininosuccinate synthetase in rat brain glial cells after striatal microinjection of immunostimulants. J Cereb Blood Flow Metab 1999, 19:898-907.
• [38]Wiesinger H: Arginine metabolism and the synthesis of nitric oxide in the nervous system. Prog Neurobiol 2001, 64:365-391.
• [39]Grady SL, Purdy JG, Rabinowitz JD, Shenk T: Argininosuccinate synthetase 1 depletion produces a metabolic state conducive to herpes simplex virus 1 infection. Proc Natl Acad Sci U S A 2013, 110:E5006-E5015.
• [40]Yokosuka M: Histological properties of the glomerular layer in the mouse accessory olfactory bulb. Exp Anim 2012, 61:13-24.
• [41]Pain F, L’heureux B, Gurden H: Visualizing odor representation in the brain: a review of imaging techniques for the mapping of sensory activity in the olfactory glomeruli. Cell Mol Life Sci 2011, 68:2689-2709.
• [42]Gray JA, McNaughton N: Comparison between the behavioural effects of septal and hippocampal lesions: A review. Neurosci Biobehav Rev 1983, 7:119-188.
• [43]Calandreau L, Jaffard R, line Desmedt A: Dissociated roles for the lateral and medial septum in elemental and contextual fear conditioning. Learn Mem 2007, 14:422-429.
• [44]Hasselmo ME: What is the function of hippocampal theta rhythm?–Linking behavioral data to phasic properties of field potential and unit recording data. Hippocampus 2005, 15:936-949.
• [45]Risold PY: The septal region. In The Rat Nervous System. Chapter 20. 3rd edition. Edited by Paxinos G. Elsevier Academic Press, Amsterdam; 2004:605-632.
• [46]Leveque M: The neuroanatomy of emotions. In Psychosurgery: New Techniques for Brain Disorders. Chapter 2. Edited by Leveque M. Springer, Switzerland; 2014:49-106.
• [47]Hikosaka O: Habenula. Scholarpedia 2007, 2:2703.
• [48]Hikosaka O: The habenula: from stress evasion to value-based decision-making. Nat Rev Neurosci 2010, 11:503-513.
• [49]Sourani D, Eitan R, Gordon N, Goelman G: The habenula couples the dopaminergic and the serotonergic systems: application to depression in Parkinson’s disease. Eur J Neurosci 2012, 36:2822-2829.
• [50]Strazielle N, Ghersi-Egea JF: Choroid plexus in the central nervous system: biology and physiopathology. J Neuropathol Exp Neurol 2000, 59:561-574.
• [51]Lehtinen MK, Bjornsson CS, Dymecki SM, Gilbertson RJ, Holtzman DM, Monuki ES: The choroid plexus and cerebrospinal fluid: emerging roles in development, disease, and therapy. J Neurosci 2013, 33:17553-17559.
• [52]Greely HT, Cho MK, Hogle LF, Satz DM: Thinking about the human neuron mouse. Am J Bioeth 2007, 7:27-40.
• [53]The Gene Expression Nervous System Atlas (GENSAT) [http://www.gensat.org/index.html]
• [54]Cheng PN, Lam TL, Lam WM, Tsui SM, Cheng AW, Lo WH, et al.: Pegylated recombinant human arginase (rhArg-peg5,000mw) inhibits the in vitro and in vivo proliferation of human hepatocellular carcinoma through arginine depletion. Cancer Res 2007, 67:309-317.
• [55]Feun L, You M, Wu CJ, Kuo MT, Wangpaichitr M, Spector S, et al.: Arginine deprivation as a targeted therapy for cancer. Curr Pharm Des 2008, 14:1049-1057.
• [56]Kung IT, Chan SK, Fung KH: Fine-needle aspiration in hepatocellular carcinoma. Combined cytologic and histologic approach. Cancer 1991, 67:673-680.