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Regulation of glycosylphosphatidylinositol-anchored proteins and GPI-phospholipase D in a c-Myc transgenic mouse model of hepatocellular carcinoma and human HCC

  • Maria Stella Ritorto , Heidrun Rhode , Arndt Vogel and Jürgen Borlak EMAIL logo
Published/Copyright: May 26, 2016

Abstract

Recent research implicated glycosylphosphatidylinositol-anchored proteins (GPI-AP) and GPI-specific phospholipase D (GPI-PLD) in the pathogenesis of fatty liver disease and hepatocellular carcinoma (HCC). Given that c-Myc is frequently amplified in HCC, we investigated their regulation in a c-Myc transgenic disease model of liver cancer and HCC patient samples. Whole genome scans defined 54 significantly regulated genes coding for GPI-AP of which 29 and 14 were repressed in expression in transgenic tumors and steatotic human hepatocyte cultures, respectively, to influence lipid-mediated signal transduction, extracellular matrix and immunity pathways. Analysis of gene specific promoter revealed >95% to carry c-Myc binding sites thus establishing a link between c-Myc activity and transcriptional response. Alike, serum GPI-PLD activity was increased 4-fold in transgenic mice; however its tissue activity was reduced by 70%. The associated repression of the serine/threonine phosphatase 2A (PP2A), i.e. a key player of c-Myc proteolysis, indicates co-ordinate responses aimed at impairing tissue GPI-PLD anti-proliferative activities. Translational research identified >4-fold increased GPI-PLD serum protein expression though enzyme activities were repressed by 60% in NASH and HCC patients. Taken collectively, c-Myc influences GPI-AP signaling transcriptionally and posttranslational and represses GPI-AP anti-proliferative signaling in tumors. The findings broaden the perspective of molecular targeted therapies and disease monitoring.

Acknowledgments

The authors thank Dr. Roman Halter for maintaining c-Myc transgenic mice and Dr. Tatiana Meier and Kerstin Wiesner for support in the microarray studies.

  1. Funding: German Federal Ministry of Education and Research (BMBF), (“The Virtual Liver Network”, Grant/Award Number: ‘031 6154’).

References

Armesto, J., Hannappel, E., Leopold, K., Fischer, W., Bublitz, R., Langer, L., Cumme, G.A., and Horn, A. (1996). Microheterogeneity of the hydrophobic and hydrophilic part of the glycosylphosphatidylinositol anchor of alkaline phosphatase from calf intestine. Eur. J. Biochem. 238, 259–269.10.1111/j.1432-1033.1996.0259q.xSearch in Google Scholar

Arnold, H.K. and Sears, R.C. (2006). Protein phosphatase 2A regulatory subunit B56alpha associates with c-myc and negatively regulates c-myc accumulation. Mol. Cell. Biol. 26, 2832–2844.10.1128/MCB.26.7.2832-2844.2006Search in Google Scholar

Borlak, J., Chougule, A., and Singh, P.K. (2014). How useful are clinical liver function tests in in vitro human hepatotoxicity assays? Toxicol. In Vitro 28, 784–795.10.1016/j.tiv.2014.03.006Search in Google Scholar

Borlak, J., Dangers, M., and Thum, T. (2002). Aroclor 1254 modulates gene expression of nuclear transcription factors: implications for albumin gene transcription and protein synthesis in rat hepatocyte cultures. Toxicol. Appl. Pharmacol. 181, 79–88.10.1006/taap.2002.9392Search in Google Scholar

Borlak, J., Hock, A., Hansen, T., and Richter, E. (2003). DNA adducts in cultures of polychlorinated biphenyl-treated human hepatocytes. Toxicol. Appl. Pharmacol. 188, 81–91.10.1016/S0041-008X(02)00075-3Search in Google Scholar

Bowman, E.P., Uhlinger, D.J., and Lambeth, J.D. (1993). Neutrophil phospholipase D is activated by a membrane-associated Rho family small molecular weight GTP-binding protein. J. Biol. Chem. 268, 21509–21512.10.1016/S0021-9258(20)80570-5Search in Google Scholar

Chalasani, N., Vuppalanchi, R., Raikwar, N.S., and Deeg, M.A. (2006). Glycosylphosphatidylinositol-specific phospholipase d in nonalcoholic fatty liver disease: a preliminary study. J. Clin. Endocrinol. Metab. 91, 2279–2285.10.1210/jc.2006-0075Search in Google Scholar

Cheng, M., Zhi, K., Gao, X., He, B., Li, Y., Han, J., Zhang, Z., and Wu, Y. (2013). Activation of cellular immunity and marked inhibition of liver cancer in a mouse model following gene therapy and tumor expression of GM-SCF, IL-21, and Rae-1. Mol. Cancer 12, 166.10.1186/1476-4598-12-166Search in Google Scholar

Ciribilli, Y., Singh, P., Spanel, R., Inga, A., and Borlak, J. (2015). Decoding c-Myc networks of cell cycle and apoptosis regulated genes in a transgenic mouse model of papillary lung adenocarcinomas. Oncotarget 6, 31569–31592.10.18632/oncotarget.5035Search in Google Scholar

Dalemans, W., Perraud, F., Le Meur, M., Gerlinger, P., Courtney, M., and Pavirani, A. (1990). Heterologous protein expression by transimmortalized differentiated liver cell lines derived from transgenic mice (hepatomas/α1 antitrypsin/ONC mouse). Biologicals 18, 191–198.10.1016/1045-1056(90)90006-LSearch in Google Scholar

Dang, C.V. (2012). MYC on the path to cancer. Cell 149, 22–35.10.1016/j.cell.2012.03.003Search in Google Scholar

Deeg, M.A., Bierman, E.L., and Cheung, M.C. (2001). GPI-specific phospholipase D associates with an apoA-I- and apoA-IV-containing complex. J. Lipid Res. 42, 442–451.10.1016/S0022-2275(20)31669-2Search in Google Scholar

Dolezal, S., Hester, S., Kirby, P.S., Nairn, A., Pierce, M., and Abbott, K.L. (2014). Elevated levels of glycosylphosphatidylinositol (GPI) anchored proteins in plasma from human cancers detected by C. septicum α toxin. Cancer Biomark. 14, 55–62.10.3233/CBM-130377Search in Google Scholar PubMed PubMed Central

Dunn, J.C., Tompkins, R.G., and Yarmush, M.L. (1991). Long-term in vitro function of adult hepatocytes in a collagen sandwich configuration. Biotechnol. Prog. 7, 237–245.10.1021/bp00009a007Search in Google Scholar PubMed

El-Serag, H.B. and Rudolph, K.L. (2007). Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 132, 2557–2576.10.1053/j.gastro.2007.04.061Search in Google Scholar PubMed

Eppig, J.T., Blake, J.A., Bult, C.J., Kadin, J.A., Richardson, J.E., and Mouse Genome Database Group (2007). The mouse genome database (MGD): new features facilitating a model system. Nucleic Acids Res. 35 (Database issue), D630–637.10.1093/nar/gkl940Search in Google Scholar PubMed PubMed Central

Fang, Y., Vilella-Bach, M., Bachmann, R., Flanigan, A., and Chen, J. (2001). Phosphatidic acid-mediated mitogenic activation of mTOR signaling. Science 294, 1942–1945.10.1126/science.1066015Search in Google Scholar PubMed

Foster, D.A. (2006). Phospholipase D survival signals as a therapeutic target in cancer. Curr. Signal Transduct. Ther. 1, 295–303.10.2174/157436206778226941Search in Google Scholar

Foster, D.A. and Xu, L. (2003). Phospholipase D in cell proliferation and cancer. Mol. Cancer Res. 1, 789–800.Search in Google Scholar

Foster, L.J., Rudich, A., Talior, I., Patel, N., Huang, X., Furtado, L.M., Bilan, P.J., Mann, M., and Klip, A. (2006). Insulin-dependent interactions of proteins with GLUT4 revealed through stable isotope labeling by amino acids in cell culture (SILAC). J. Proteome Res. 5, 64–75.10.1021/pr0502626Search in Google Scholar PubMed

Frau, M., Biasi, F., Feo, F., and Pascale, R.M. (2010). Prognostic markers and putative therapeutic targets for hepatocellular carcinoma. Mol. Aspects Med. 31, 179–193.10.1016/j.mam.2010.02.007Search in Google Scholar PubMed

Gomez-Cambronero, J. (2014). Phospholipase D in cell signaling: from a myriad of cell functions to cancer growth and metastasis. J. Biol. Chem. 289, 22557–22566.10.1074/jbc.R114.574152Search in Google Scholar

Hammond, S.M., Jenco, J.M., Nakashima, S., Cadwallader, K., Gu, Q., Cook, S., Nozawa, Y., Prestwich, G.D., Frohman, M.A., and Morris, A.J. (1997). Characterization of two alternately spliced forms of phospholipase D1. Activation of the purified enzymes by phosphatidylinositol 4,5-bisphosphate, ADP-ribosylation factor, and Rho family monomeric GTP-binding proteins and protein kinase C-α. J. Biol. Chem. 272, 3860–3868.10.1074/jbc.272.6.3860Search in Google Scholar

Hui, L., Rodrik, V., Pielak, R.M., Knirr, S., Zheng, Y., and Foster, D.A. (2005). mTOR-dependent suppression of protein phosphatase 2A is critical for phospholipase D survival signals in human breast cancer cells. J. Biol. Chem. 280, 35829–35835.10.1074/jbc.M504192200Search in Google Scholar

Hunecke, D., Spanel, R., Länger, F., Nam, S.W., and Borlak, J. (2012). MYC-regulated genes involved in liver cell dysplasia identified in a transgenic model of liver cancer. J. Pathol. 228, 520–533.10.1002/path.4059Search in Google Scholar

Jiang, J., Nilsson-Ehle, P., and Xu, N. (2006). Influence of liver cancer on lipid and lipoprotein metabolism. Lipids Health Dis. 5, 4.10.1186/1476-511X-5-4Search in Google Scholar

Johnson, J.M., Castle, J., Garrett-Engele, P., Kan, Z., Loerch, P.M., Armour, C.D., Santos, R., Schadt, E.E., Stoughton, R., and Shoemaker, D.D. (2003). Genome-wide survey of human alternative pre-mRNA splicing with exon junction microarrays. Science 302, 2141–2144.10.1126/science.1090100Search in Google Scholar

Jones, D.R. and Varela-Nieto, I. (1998). The role of glycosyl-phosphatidylinositol in signal transduction. Int. J. Biochem. Cell Biol. 30, 313–326.10.1016/S1357-2725(97)00144-1Search in Google Scholar

Joseph, T., Bryant, A., Frankel, P., Wooden, R., Kerkhoff, E., Rapp, U.R., and Foster, D.A. (2002). Phospholipase D overcomes cell cycle arrest induced by high-intensity Raf signaling. Oncogene 21, 3651–3658.10.1038/sj.onc.1205380Search in Google Scholar PubMed

Junttila, M.R. and Westermarck, J. (2008). Mechanisms of MYC stabilization in human malignancies. Cell Cycle 7, 592–596.10.4161/cc.7.5.5492Search in Google Scholar PubMed

Kerkhoff, E., Houben, R., Loffler, S., Troppmair, J., Lee, J.E., and Rapp, U.R. (1998). Regulation of c-myc expression by Ras/Raf signalling. Oncogene 16, 211–216.10.1038/sj.onc.1201520Search in Google Scholar PubMed

Liscovitch, M., Czarny, M., Fiucci, G., and Tang, X. (2000). Phospholipase D: molecular and cell biology of a novel gene family. Biochem. J. 345 (Pt 3), 401–415.10.1042/bj3450401Search in Google Scholar

Low, M.G. and Huang, K.S. (1993). Phosphatidic acid, lysophosphatidic acid, and lipid A are inhibitors of glycosylphosphatidylinositol-specific phospholipase D. Specific inhibition of a phospholipase by product analogues? J. Biol. Chem. 268, 8480–8490.10.1016/S0021-9258(18)52900-8Search in Google Scholar

Low, M.G. and Stütz, P. (1999). Inhibition of the plasma glycosylphosphatidylinositol-specific phospholipase D by synthetic analogs of lipid A and phosphatidic acid. Arch. Biochem. Biophys. 371, 332–339.10.1006/abbi.1999.1436Search in Google Scholar

Mostafavi, S., Ray, D., Warde-Farley, D., Grouios, C., and Morris, Q. (2008). GeneMANIA: a real-time multiple association network integration algorithm for predicting gene function. Genome Biol. 9 (Suppl. 1), S4.10.1186/gb-2008-9-s1-s4Search in Google Scholar

Ooi, K., Shiraki, K., Sakurai, Y., Morishita, Y., and Nobori, T. (2005). Clinical significance of abnormal lipoprotein patterns in liver diseases. Int. J. Mol. Med. 15, 655–660.10.3892/ijmm.15.4.655Search in Google Scholar

Pelengaris, S. and Khan, M. (2003). The many faces of c-MYC. Arch. Biochem. Biophys. 416, 129–136.10.1016/S0003-9861(03)00294-7Search in Google Scholar

Pierleoni, A., Martelli, P.L., and Casadio, R. (2008). PredGPI: a GPI-anchor predictor. BMC Bioinf. 9, 392.10.1186/1471-2105-9-392Search in Google Scholar

Plummer, G., Perreault, K.R., Holmes, C.F., and Posse De Chaves, E.I. (2005). Activation of serine/threonine protein phosphatase-1 is required for ceramide-induced survival of sympathetic neurons. Biochem. J. 385, 685–693.10.1042/BJ20040929Search in Google Scholar

Powner, D.J. and Wakelam, M.J. (2002). The regulation of phospholipase D by inositol phospholipids and small GTPases. FEBS Lett. 531, 62–64.10.1016/S0014-5793(02)03410-5Search in Google Scholar

Rhode, H., Hoffmann-Blume, E., Schilling, K., Gehrhardt, S., Gohlert, A., Buttner, A., Bublitz, R.R., Cumme, G.A., and Horn, A. (1995). Glycosylphosphatidylinositol-alkaline phosphatase from calf intestine as substrate for glycosylphosphatidylinositol-specific phospholipases--microassay using hydrophobic chromatography in pipet tips. Anal. Biochem. 231, 99–108.10.1006/abio.1995.1508Search in Google Scholar PubMed

Rhode, H., Lopatta, E., Schulze, M., Pascual, C., Schulze, H.P., Schubert, K., Schubert, H., Reinhart, K., and Horn, A. (1999). Glycosylphosphatidylinositol-specific phospholipase D in blood serum: is the liver the only source of the enzyme? Clin. Chim. Acta 281, 127–145.10.1016/S0009-8981(98)00218-6Search in Google Scholar

Rhode, H., Schulze, M., Cumme, G.A., Gohlert, A., Blume, E., Bublitz, R., Schilling, K., and Horn, A. (2000). Glycosylphosphatidylinositol-specific phospholipase D of human serum--activity modulation by naturally occurring amphiphiles. Biol. Chem. 381, 471–485.10.1515/BC.2000.062Search in Google Scholar PubMed

Ritorto, M.S. and Borlak, J. (2011). Combined serum and tissue proteomic study applied to a c-Myc transgenic mouse model of hepatocellular carcinoma identified novel disease regulated proteins suitable for diagnosis and therapeutic intervention strategies. J. Proteome Res. 10, 3012–3030.10.1021/pr101207tSearch in Google Scholar PubMed

Rodrik, V., Zheng, Y., Harrow, F., Chen, Y., and Foster, D.A. (2005). Survival signals generated by estrogen and phospholipase D in MCF-7 breast cancer cells are dependent on Myc. Mol. Cell. Biol. 25, 7917–7925.10.1128/MCB.25.17.7917-7925.2005Search in Google Scholar PubMed PubMed Central

Rodrik, V., Gomes, E., Hui, L., Rockwell, P., and Foster, D.A. (2006). Myc stabilization in response to estrogen and phospholipase D in MCF-7 breast cancer cells. FEBS Lett. 580, 5647–5652.10.1016/j.febslet.2006.09.013Search in Google Scholar PubMed PubMed Central

Sernova, N.V. (2006). Evaluation of programs for the prediction of GPI-anchor in proteins. Report of Practical Training performed at Swiss-Prot group, Swiss Institute of Bioinformatics. Available at: http://www.mpb.unige.ch/reports/rep_Natalia_Sernova.pdf2015, 1–57.Search in Google Scholar

Shanley, T.P., Vasi, N., Denenberg, A., and Wong, H.R. (2001). The serine/threonine phosphatase, PP2A: endogenous regulator of inflammatory cell signaling. J. Immunol. 166, 966–972.10.4049/jimmunol.166.2.966Search in Google Scholar PubMed

Shi, M., Zheng, Y., Garcia, A., Xu, L., and Foster, D.A. (2007). Phospholipase D provides a survival signal in human cancer cells with activated H-Ras or K-Ras. Cancer Lett. 258, 268–275.10.1016/j.canlet.2007.09.003Search in Google Scholar PubMed PubMed Central

Song, Y., Wu, J., Oyesanya, R.A., Lee, Z., Mukherjee, A., and Fang, X. (2009). Sp-1 and c-Myc mediate lysophosphatidic acid-induced expression of vascular endothelial growth factor in ovarian cancer cells via a hypoxia-inducible factor-1-independent mechanism. Clin. Cancer Res. 15, 492–501.10.1158/1078-0432.CCR-08-1945Search in Google Scholar PubMed PubMed Central

Tang, J.H., He, W.J., Huang, H., Tan, C.C., Duan, Q., Wang, K.J., Yuan, X.Y., and Zhu, X.J. (2009). Important roles of glycosylphosphatidylinositol (GPI)-specific phospholipase D and some GPI-anchored proteins in the pathogenesis of hepatocellular carcinoma. Clin. Biochem. 42, 400–407.10.1016/j.clinbiochem.2008.11.019Search in Google Scholar PubMed

Tsujioka, H., Misumi, Y., Takami, N., and Ikehara, Y. (1998). Posttranslational modification of glycosylphosphatidylinositol (GPI)-specific phospholipase D and its activity in cleavage of GPI anchors. Biochem. Biophys. Res. Commun. 251, 737–743.10.1006/bbrc.1998.9542Search in Google Scholar PubMed

Walker, E.J., Rosenberg, S.A., Wands, J.R., and Kim, M. (2011). Role of Raf kinase inhibitor protein in hepatocellular carcinoma. For. Immunopathol. Dis. Therap. 2, 195–204.10.1615/ForumImmunDisTher.v2.i2.110Search in Google Scholar PubMed PubMed Central

Warde-Farley, D., Donaldson, S.L., Comes, O., Zuberi, K., Badrawi, R., Chao, P., Franz, M., Grouios, C., Kazi, F., Lopes, C.T., et al. (2010). The GeneMANIA prediction server: biological network integration for gene prioritization and predicting gene function. Nucleic Acids Res. 38, W214–20.10.1093/nar/gkq537Search in Google Scholar PubMed PubMed Central

Yang, H.J., Hsu, C.L., Yang, J.Y., and Yang, W.Y. (2012). Monodansylpentane as a blue-fluorescent lipid-droplet marker for multi-color live-cell imaging. PLoS One 7, e32693.10.1371/journal.pone.0032693Search in Google Scholar PubMed PubMed Central

Zheng, Y., Rodrik, V., Toschi, A., Shi, M., Hui, L., Shen, Y., and Foster, D.A. (2006). Phospholipase D couples survival and migration signals in stress response of human cancer cells. J. Biol. Chem. 281, 15862–15868.10.1074/jbc.M600660200Search in Google Scholar PubMed

Zhou, M., Lucas, D.A., Chan, K.C., Issaq, H.J., Petricoin, E.F., 3rd, Liotta, L.A., Veenstra, T.D., and Conrads, T.P. (2004). An investigation into the human serum “interactome”. Electrophoresis 25, 1289–1298.10.1002/elps.200405866Search in Google Scholar PubMed


Supplemental Material:

The online version of this article (DOI: 10.1515/hsz-2016-0133) offers supplementary material, available to authorized users.


Received: 2016-2-5
Accepted: 2016-5-24
Published Online: 2016-5-26
Published in Print: 2016-11-1

©2016 Walter de Gruyter GmbH, Berlin/Boston

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