Abstract
Glioma is the most common type of brain tumor, and has the worst prognosis in human malignancy. Experimental evidence suggests that the use of high concentrations of various amino acids may perturb neoplastic cell growth. Thus, the aim of this study was to investigate whether essential amino acids can alter the growth and proliferation of glioma cells. Studies were performed using C6 rat glioma cell lines. High concentration of l-leucine induced growth arrest of glioma cell lines. Terminal transferase uridyl nick end labeling assay and cell cycle analysis showed that the effect of l-leucine on glioma cells growth was not cytotoxic, but rather cytostatic. Additionally, the extracellular signal-regulated protein kinase was activated in l-leucine-treated glioma cells, and inhibition of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1 (MEK) enhanced the effect of l-leucine on glioma cell growth. These data suggest that high concentration l-leucine combined with inhibition of MEK is a potential strategy for glioma cell growth arrest.
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References
Alao JP (2007) The regulation of cyclin D1 degradation: roles in cancer development and the potential for therapeutic invention. Mol Cancer 6:24
Avruch J, Long X, Ortiz-Vega S, Rapley J, Papageorgiou A, Dai N (2009) Amino acid regulation of TOR complex 1. Am J Physiol Endocrinol Metab 296:E592–E602
Christensen HN (1990) Role of amino acid transport and countertransport in nutrition and metabolism. Physiol Rev 70:43–77
Cohen A, Hall MN (2009) An amino acid shuffle activates mTORC1. Cell 136:399–400
Goberdhan DC (2010) Intracellular amino acid sensing and mTORC1-regulated growth: new ways to block an old target? Curr Opin Investig Drugs 11:1360–1367
Hara K, Yonezawa K, Weng QP, Kozlowski MT, Belham C, Avruch J (1998) Amino acid sufficiency and mTOR regulate p70 S6 kinase and eIF-4E BP1 through a common effector mechanism. J Biol Chem 273:14484–14494
Hidayat S, Yoshino K, Tokunaga C, Hara K, Matsuo M, Yonezawa K (2003) Inhibition of amino acid-mTOR signaling by a leucine derivative induces G1 arrest in Jurkat cells. Biochem Biophys Res Commun 301:417–423
Ishizuka Y, Kakiya N, Nawa H, Takei N (2008) Leucine induces phosphorylation and activation of p70S6K in cortical neurons via the system L amino acid transporter. J Neurochem 106:934–942
Kanai Y, Segawa H, Miyamoto K, Uchino H, Takeda E, Endou H (1998) Expression cloning and characterization of a transporter for large neutral amino acids activated by the heavy chain of 4F2 antigen (CD98). J Biol Chem 273:23629–23632
Lee MY, Jo SD, Lee JH, Han HJ (2008) l-leucine increases [3H]-thymidine incorporation in chicken hepatocytes: involvement of the PKC, PI3K/Akt, ERK1/2, and mTOR signaling pathways. J Cell Biochem 105:1410–1419
Leung-Pineda V, Pan Y, Chen H, Kilberg MS (2004) Induction of p21 and p27 expression by amino acid deprivation of HepG2 human hepatoma cells involves mRNA stabilization. Biochem J 379:79–88
Lind DS (2004) Arginine and cancer. J Nutr 134:2837S–2841S
Mastroberardino L, Spindler B, Pfeiffer R, Skelly PJ, Loffing J, Shoemaker CB, Verrey F (1998) Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family. Nature 395:288–291
Mitsubuchi H, Owada M, Endo F (2005) Markers associated with inborn errors of metabolism of branched-chain amino acids and their relevance to upper levels of intake in healthy people: an implication from clinical and molecular investigations on maple syrup urine disease. J Nutr 135:1565S–1570S
Nawashiro H, Otani N, Shinomiya N, Fukui S, Ooigawa H, Shima K, Matsuo H, Kanai Y, Endou H (2006) L-type amino acid transporter 1 as a potential molecular target in human astrocytic tumors. Int J Cancer 119:484–492
Nicklin P, Bergman P, Zhang B, Triantafellow E, Wang H, Nyfeler B, Yang H, Hild M, Kung C, Wilson C, Myer VE, MacKeigan JP, Porter JA, Wang YK, Cantley LC, Finan PM, Murphy LO (2009) Bidirectional transport of amino acids regulates mTOR and autophagy. Cell 136:521–534
Nishio Y, Kakizoe T, Ohtani M, Sato S, Sugimura T, Fukushima S (1986) l-isoleucine and l-leucine: tumor promoters of bladder cancer in rats. Science 231:843–845
Peyrollier K, Hajduch E, Blair AS, Hyde R, Hundal HS (2000) l-leucine availability regulates phosphatidylinositol 3-kinase, p70 S6 kinase and glycogen synthase kinase-3 activity in L6 muscle cells: evidence for the involvement of the mammalian target of rapamycin (mTOR) pathway in the l-leucine-induced up-regulation of system A amino acid transport. Biochem J 350:361–368
Sang J, Lim YP, Panzica M, Finch P, Thompson NL (1995) TA1, a highly conserved oncofetal complementary DNA from rat hepatoma, encodes an integral membrane protein associated with liver development, carcinogenesis, and cell activation. Cancer Res 55:1152–1159
Scholl-Bürgi S, Haberlandt E, Heinz-Erian P, Deisenhammer F, Albrecht U, Sigl SB, Rauchenzauner M, Ulmer H, Karall D (2008) Amino acid cerebrospinal fluid/plasma ratios in children: influence of age, gender, and antiepileptic medication. Pediatrics 121:e920–e926
Scott CB, Scarantino C, Urtasun R, Movsas B, Jones CU, Simpson JR, Fischbach AJ, Curran WJ Jr (1998) Validation and predictive power of Radiation Therapy Oncology Group (RTOG) recursive partitioning analysis classes for malignant glioma patients: a report using RTOG 90-06. Int J Radiat Oncol Biol Phys 40:51–55
Seger R, Krebs EG (1995) The MAPK signaling cascade. FASEB J 9:726–735
Sheen JH, Zoncu R, Kim D, Sabatini DM (2011) Defective regulation of autophagy upon leucine deprivation reveals a targetable liability of human melanoma cells in vitro and in vivo. Cancer Cell 19:613–628
Souba WW (1993) Glutamine and cancer. Ann Surg 218:715–728
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO, European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups, National Cancer Institute of Canada Clinical Trials Group (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996
Wakshlag JJ, Kallfelz FA, Wakshlag RR, Davenport GM (2006) The effects of branched-chain amino acids on canine neoplastic cell proliferation and death. J Nutr 136:2007S–2010S
Wolf DA, Wang S, Panzica MA, Bassily NH, Thompson NL (1996) Expression of a highly conserved oncofetal gene, TA1/E16, in human colon carcinoma and other primary cancers: homology to Schistosoma mansoni amino acid permease and Caenorhabditis elegans gene products. Cancer Res 56:5012–5022
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We thank Akiko Yano for technical assistance in preparation of data for the manuscript.
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Takeuchi, S., Nawashiro, H., Wada, K. et al. l-Leucine induces growth arrest and persistent ERK activation in glioma cells. Amino Acids 43, 717–724 (2012). https://doi.org/10.1007/s00726-011-1122-9
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DOI: https://doi.org/10.1007/s00726-011-1122-9