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The Effect of Antineoplastons A10 and AS2-1 and Metabolites of Sodium Phenylbutyrate on Gene Expression in Glioblastoma Multiforme

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DOI: 10.4236/jct.2014.510099    2,868 Downloads   3,387 Views   Citations

ABSTRACT

Antineoplastons are peptide and amino acid derivatives that occur naturally in the human body. They inhibit the growth of neoplastic cells without growth inhibition of normal cells. Phenylacetylglutaminate (PG) is an active ingredient of antineoplastons A10 and AS2-1 (ANP) and is also a metabolic by-product of phenylbutyrate (PB). The formulation of antineoplaston AS2-1 is a 4:1 mixture of phenylacetate (PN) and PG. Antineoplaston A10 is a 4:1 mixture of PG and isoPG. This study investigates the molecular mechanism of action of PG and PN. The Human U87 glioblastoma (GBM) cell line was used as the model system in this study. A total human gene array screen using the Affymetrix Human Genome plus 2.0 oligonucleotide arrays was performed using mRNA derived from U87 cells exposed to PG and PN. Pathway analysis was performed to allow the visualization of effect on metabolic pathways and gene interaction networks. Our preliminary results indicate that PG and PN interrupt signal transduction in RAS/MAPK/ERK and PI3K/AKT/PTEN pathways, interfere with cell cycle, decrease metabolism and promote apoptosis in human U87 GBM cells. The effect on multiple cellular pathways and targets, suggests that ANP and PB are promising candidates for clinical studies in GBM.

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Burzynski, S. and Patil, S. (2014) The Effect of Antineoplastons A10 and AS2-1 and Metabolites of Sodium Phenylbutyrate on Gene Expression in Glioblastoma Multiforme. Journal of Cancer Therapy, 5, 929-945. doi: 10.4236/jct.2014.510099.

References

[1] Burzynski, S. (1968) Investigations on Amino Acids and Peptides in Blood Serum of Healthy People and Patients with Chronic Renal Insufficiency. Medical Academy, Lublin.
[2] Burzynski, S. (1969) Investigations on Unknown Ninhydrin-Reacting Substances in Human Blood Serum. I. Attempts at Identification of Three Such Substances. Experientia, 25, 490-491.
http://dx.doi.org/10.1007/BF01900774
[3] Burzynski, S.R. (1973) Biologically Active Peptides in Human Urine: I. Isolation of a Group of Medium-Sized Peptides. Physiological Chemistry and Physics, 5, 437-447.
[4] Burzynski, S.R., Ungar, A.L. and Lubanski, E. (1974) Biologically Active Peptides in Human Urine: II. Effect on Intestinal Smooth Muscle and Heart. Physiological Chemistry and Physics, 6, 457-468.
[5] Burzynski, S.R., Loo, T.L., Ho, D.H., Rao, P.N., Georgiades, G. and Kratzenstein, H. (1978) Biologically Active Peptides in Human Urine: III. Inhibitors of the Growth of Leukemia, Osteosarcoma and HeLa Cells. Physiological Chemistry and Physics, 8, 13-22.
[6] Burzynski, S.R. (1976) Antineoplastons: Biochemical Defense against Cancer. Physiological Chemistry and Physics, 8, 275-279.
[7] Burzynski, S.R. (1986) Antineoplastons—History of the Research (I). Drugs under Experimental and Clinical Research, 12, 1-9.
[8] Burzynski, S.R. (2004) The Present State of Antineoplaston Research (1). Integrative Cancer Therapies, 3, 47-58. http://dx.doi.org/10.1177/1534735403261964
[9] Bartek, J., Ng, K., Fischer, W., Carter, B. and Chen, C. (2012) Key Concepts in Glioblastoma Therapy. Journal of Neurology, Neurosurgery Psychiatry, 83, 753-760. http://dx.doi.org/10.1136/jnnp-2011-300709
[10] Burzynski, S.R. (1995) Potential of Antineoplastons in Diseases of Old Age. Drugs Aging, 7, 157-167. http://dx.doi.org/10.2165/00002512-199507030-00001
[11] Ostrom, Q.T., Gittleman, H.R., Farah, P., Ondracek, A., Chen, Y., Wolinsky, Y., Stroup, N.E., Kruchko, C. and Barnholtz-Sloan, J.S. (2013) CBTRUS Statistical Report: Primary Brain and Central Nervous System Tumors Diagnosed in the United States 2006-2010. Neuro-Oncology, 15, ii1-ii56.
http://dx.doi.org/10.1093/neuonc/not151
[12] Newton, H.B. (1994) Primary Brain Tumors: Review of Etiology, Diagnosis, and Treatment. American Family Physician, 49, 787-797.
[13] Buckner, J.C., Brown, P.D., O’Neill, B.P., Meyer, F.B., Wetmore, C.J. and Uhm, J.H. (2007) Central Nervous System Tumors. Mayo Clinic Proceedings, 82, 1271-1286.
http://dx.doi.org/10.4065/82.10.1271
[14] CGAR Network (2008) Comprehensive Genomic Characterization Defines Human Glioblastoma Genes and Core Pathways. Nature, 455, 1061-1068. http://dx.doi.org/10.1038/nature07385
[15] Sturm, D., Witt, H., Hovestadt, V., Khuong-Quang, D.A., Jones, D.T.W., Konermann, C., et al. (2012) Hotspot Mutations in H3F3A and IDH1 Define Distinct Epigenetic and Biological Subgroups of Glioblastoma. Cancer Cell, 22, 425-437. http://dx.doi.org/10.1016/j.ccr.2012.08.024
[16] Kim, Y.W., Koul, D., Kim, S.H., Lucio-Eterovic, A.K., Freire, P.R., Yao, J., Wang, J., Almeida, J.S., Aldape, K. and Alfred Yung, W.K. (2013) Identification of Prognostic Gene Signatures of Glioblastoma: A Study Based on TCGA Data Analysis. Neuro-Oncology, 15, 829-839.
http://dx.doi.org/10.1093/neuonc/not024
[17] Maitland, M.L. and Schilsky, R.L. (2011) Clinical Trials in the Era of Personalized Oncology. CA: A Cancer Journal for Clinicians, 61, 365-381. http://dx.doi.org/10.3322/caac.20135
[18] de Tayrac, M., Aubry, M., Saikali, S., Etcheverry, A., Surbled, C., Guenot, F., et al. (2011) A 4-Gene Signature Associated with Clinical Outcome in High-Grade Gliomas. Clinical Cancer Research, 17, 317-327. http://dx.doi.org/10.1158/1078-0432.CCR-10-1126
[19] Burzynski, S.R. (2006) Treatments for Astrocytic Tumors in Children: Current and Emerging Strategies. Pediatric Drugs, 8, 167-168. http://dx.doi.org/10.2165/00148581-200608030-00003
[20] Burzynski, S.R., Janicki, T.J., Burzynski, G.S. and Marszalek, A. (2014) The Response and Survival of Children with Recurrent Diffuse Intrinsic Pontine Glioma on Phase II Study of Antineoplastons A10 and AS2-1 in Patients with Brainstem Glioma. Child’s Nervous System.
http://link.springer.com/article/10.1007/s00381-014-2401-z
[21] Burzynski, S.R., Janicki, T.J., Burzynski, G.S. and Marszalek, A. (2014) A Phase II Study of Antineoplastons A10 and AS2-1 in Children with High-Grade Glioma. Final Report (Protocol BT-06) and Review of Recent Trials. Journal of Cancer Therapy, 5, 565-577.
http://dx.doi.org/10.4236/jct.2014.56065
[22] Burzynski, S.R., Janicki, T.J. and Burzynski, G.S. (2014) A Phase II Study of Antineoplastons A10 and AS2-1 in Adult Patients with Recurrent Glioblastoma Multiforme. Final Report (Protocol BT-21). Journal of Cancer Therapy, in press.
[23] Burzynski, S.R., Janicki, T.J., Burzynski, G.S. and Marszalek, A. (2014) A Phase II Study of Antineoplastons A10 and AS2-1 in Children with Recurrent Refractory or Progressive Primary Brain Tumors. Finalr Report (Protocol BT-22). Journal of Cancer Therapy, in press.
[24] Burzynski, S.R., Janicki, T.J. and Burzynski, G.S. (2014) Recurrent Glioblastoma Multiforme, a Strategy for Long Term Survival. Journal of Cancer Therapy, in press.
[25] Brusilow, S.W., Danney, M., Waber, L.J., Batshaw, M., Burton, B., Levitsky, L., Roth, K., McKeethren, C. and Ward, J. (1984) Treatment of Episodic Hyperammonemia in Children with Inborn Errors of Urea Synthesis. New England Journal of Medicine, 310, 1630-1634.
http://dx.doi.org/10.1056/NEJM198406213102503
[26] Iannitti, T. and Palmieri, B. (2011) Clinical and Experimental Applications of Sodium Phenylbutyrate. Drugs in R & D, 11, 227-249. http://dx.doi.org/10.2165/11591280-000000000-00000
[27] Burzynski, S.R. (2006) Targeted Therapy for Brain Tumors. In: Yang, A.V., Ed., Brain Cancer Therapy and Surgical Interventions, Horizons in Cancer Research, Vol. 27, Nova Science Publishers, Inc., Hauppauge, 77-11.
[28] Patil, S.S., Burzynski, S.R., Mrowczynski, E., Grela, K. and Chittur, S.V. (2012) Phenylacetylglutaminate and Phenylacetate in Combination Upregulate VDUP1, Cause Cell Cycle Blockade and Apoptosis in U87 Glioblastoma Cells. Journal of Cancer Therapy, 3, 192-200.
http://dx.doi.org/10.4236/jct.2012.33028
[29] Malumbres, M. and Barbacid, M. (2003) RAS Oncogenes: The First 30 Years. Nature Reviews Cancer, 3, 459-465. http://dx.doi.org/10.1038/nrc1097
[30] Newton, H.B. (2004) Molecular Neuro-Oncology and Development of Targeted Therapeutic Strategies for Brain Tumors. Part 2: PI3K/Akt/PTEN, mTOR, SHH/PTCH and Angiogenesis. Expert Review of Anticancer Therapy, 4, 105-128. http://dx.doi.org/10.1586/14737140.4.1.105
[31] Fujii, T., Nakamura, A.M., Yokoyama, G., Yamaguchi, M., Tayama, K., Miwa, K., et al. (2005) Antineoplaston Induces G1 Arrest by PKCalpha and MAPK Pathway in SKBR-3 Breast Cancer Cells. Oncology Reports, 14, 489-494.
[32] Deschenes-Simard, X., Kottakis, F., Meloche, S. and Ferbeyre, G. (2014) ERKs in Cancer: Friends or Foes? Cancer Research, 74, 412-419. http://dx.doi.org/10.1158/0008-5472.CAN-13-2381
[33] Vivanco, I. and Sawyers, C.L. (2002) The Phosphatidylinositol 3-Kinase AKT Pathway in Human Cancer. Nature Reviews Cancer, 2, 489-501. http://dx.doi.org/10.1038/nrc839
[34] Bjornsti, M.A. and Houghton, P.J. (2004) The TOR Pathway: A Target for Cancer Therapy. Nature Reviews Cancer, 4, 335-348. http://dx.doi.org/10.1038/nrc1362
[35] Sonoda, Y., Ozawa, T., Aldape, K.D., Deen, D.F., Berger, M.S. and Pieper, R.O. (2001) Akt Pathway Activation Converts Anaplastic Astrocytoma to Glioblastoma Multiforme in a Human Astrocyte Model of Glioma. Cancer Research, 61, 6674-6678.
[36] Mills, G.B., Lu, Y. and Kohn, E.C. (2001) Linking Molecular Therapeutic to Molecular Diagnostics: Inhibition of the FRAP/RAFT/TOR Component of the PI3K Pathway Preferentially Blocks PTEN Mutant Cells in Vitro and in Vivo. Proceedings of the National Academy of Sciences of the United States of America, 98, 10031-10033. http://dx.doi.org/10.1073/pnas.191379498
[37] Nishiyama, A., Matsui, M., Iwata, S., Hirota, K., Masutani, H., Nakamura, H., Takagi, Y., Sono, H., Gon, Y. and Yodoi, J. (1999) Identification of Thioredoxin-Binding Protein-2/Vitamin D3 Up-Regulated Protein 1 as a Negative Regulator of Thioredoxin Function and Expression. Journal of Biological Chemistry, 274, 21645-21650. http://dx.doi.org/10.1074/jbc.274.31.21645
[38] Han, S.H., Jeon, J.H., Ju, H.R., Jung, U., Kim, K.Y., Yoo, H.S., et al. (2003) VDUP1 Upregulated by TGF-β1 and 1,25-Dihydroxy-Vitamin D3 Inhibits Tumor Cell Growth by Blocking Cell-Cycle Progression. Oncogene, 22, 4035-4046.http://dx.doi.org/10.1038/sj.onc.1206610
[39] Massague, J. (2004) G1 Cell-Cycle Control and Cancer. Nature, 432, 298-306.
http://dx.doi.org/10.1038/nature03094
[40] Pelengaris, S., Khan, M. and Evan, G. (2002) c-MYC: More than Just a Matter of Life and Death. Nature Reviews Cancer, 2, 764-776. http://dx.doi.org/10.1038/nrc904
[41] Brown, J.M. and Attardi, L.D. (2005) The Role of Apoptosis in Cancer Development and Treatment Response. Nature Reviews Cancer, 5, 231-237.
[42] Ghobrial, I.M., Witzig, T.E. and Adjei, A.A. (2005) Targeting Apoptosis Pathways in Cancer Therapy. CA: A Cancer Journal for Clinicians, 55, 178-194. http://dx.doi.org/10.3322/canjclin.55.3.178
[43] Hueber, A.O., Zornig, M., Lyon, D., Suda, T., Nagata, S. and Evan, G.I. (1997) Requirement for the CD95 Receptor-Ligand Pathway in c-Myc-Induced Apoptosis. Science, 278, 1305-1309.
http://dx.doi.org/10.1126/science.278.5341.1305
[44] Hockenbery, D., Nunez, G., Milliman, C., Schreiber, R.D. and Korsmeyer, S.J. (1990) Bcl-2 Is an Inner Mitochondrial Membrane Protein That Blocks Programmed Cell Death. Nature, 348, 334-336. http://dx.doi.org/10.1038/348334a0
[45] Levine, A.J. (1997) p53, the Cellular Gatekeeper for Growth and Division. Cell, 88, 323-331. http://dx.doi.org/10.1016/S0092-8674(00)81871-1
[46] el-Deiry, W.S., Harper, J.W., O’Connor, P.M., Velculescu, V.E., Canman, C.E., Jackman, J., et al. (1994) WAF1/CIP1 Is Induced in p53-Mediated G1 Arrest and Apoptosis. Cancer Research, 54, 1169-1174.
[47] Yu, J., Zhang, L., Hwang, P.M., Rago, C., Kinzler, K.W. and Vogelstein, B. (1999) Identification and Classification of p53-Regulated Genes. Proceedings of the National Academy of Sciences of the United States of America, 96, 14517-14522. http://dx.doi.org/10.1073/pnas.96.25.14517
[48] Lozano, G. and Zambetti, G.P. (2005) What Have Animal Models Taught Us about the p53 Pathway? Journal of Pathology, 205, 206-220. http://dx.doi.org/10.1002/path.1704
[49] Parsons, D.W., Jones, S., Zhang, X., Lin, J.C., Leary, R.J., Angenendt, P., et al. (2008) An Integrated Genomic Analysis of Human Glioblastoma Multiforme. Science, 321, 1807-1812.
http://dx.doi.org/10.1126/science.1164382
[50] Wick, W., Weller, M., Weiler, M., Batchelor, T., Yung, A.W.K. and Platten, M. (2011) Pathway Inhibition: Emerging Molecular Targets for Treating Glioblastoma. Neuro-Oncology, 13, 566-579. http://dx.doi.org/10.1093/neuonc/nor039
[51] Chinnaiyan, P., Kensicki, E., Bloom, G., Prabhu, A., Sarcar, B., Kahali, S., Eschrich, S., Qu, X.T., Forsyth, P. and Gillies, R. (2012) The Metabolomic Signature of Malignant Glioma Reflects Accelerated Anabolic Metabolism. Cancer Research, 72, 5878-5888.http://dx.doi.org/10.1158/0008-5472.CAN-12-1572-T
[52] Clem, B.F. and Chesney, J. (2012) Molecular Pathways: Regulation of Metabolism by RB. Clinical Cancer Research, 18, 6096-6100. http://dx.doi.org/10.1158/1078-0432.CCR-11-3164
[53] Dou, Q.P., Zhao, S., Levin, A.H., Wang, J., Helin, K. and Pardee, A.B. (1994) G1/S-Regulated E2F-Containing Protein Complexes Bind to the Mouse Thymidine Kinase Gene Promoter. Journal of Biological Chemistry, 269, 1306-1313.
[54] Tommasi, S. and Pfeifer, G.P. (1997) Constitutive Protection of E2F Recognition Sequences in the Human Thymidine Kinase Promoter during Cell Cycle Progression. Journal of Biological Chemistry, 272, 30483-30490. http://dx.doi.org/10.1074/jbc.272.48.30483
[55] Wade, M., Kowalik, T.F., Mudryj, M., Huang, E.S. and Azizkhan, J.C. (1992) E2F Mediates Dihydrofolate Reductase Promoter Activation and Multiprotein Complex Formation in Human Cytomegalovirus Infection. Molecular and Cellular Biology, 12, 4364-4374.
[56] Dang, C.V., Le, A. and Gao, P. (2009) MYC-Induced Cancer Cell Energy Metabolism and Therapeutic Opportunities. Clinical Cancer Research, 15, 6479-6483. http://dx.doi.org/10.1158/1078-0432.CCR-09-0889
[57] DeBerardinis, R.J. and Cheng, T. (2009) Q’s Next: The Diverse Functions of Glutamine in Metabolism, Cell Biology and Cancer. Oncogene, 29, 313-324. http://dx.doi.org/10.1038/onc.2009.358
[58] Wise, D.R. and Thompson, C.B. (2010) Glutamine Addiction: A New Therapeutic Target in Cancer. Trends in Biochemical Sciences, 35, 427-433. http://dx.doi.org/10.1016/j.tibs.2010.05.003
[59] Gao, P., Tchernyshyov, I., Chang, T.C., Lee, Y.S., Kita, K., Ochi, T., et al. (2009) c-Myc Suppression of miR-23a/b Enhances Mitochondrial Glutaminase Expression and Glutamine Metabolism. Nature, 458, 762-765. http://dx.doi.org/10.1038/nature07823
[60] Yuneva, M., Zamboni, N., Oefner, P., Sachidanandam, R. and Lazebnik, Y. (2007) Deficiency in Glutamine but Not Glucose Induces MYC-Dependent Apoptosis in Human Cells. Journal of Cell Biology, 178, 93-105. http://dx.doi.org/10.1083/jcb.200703099

  
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