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Expression Profile of Genes Modulated by Aloe emodin in Human U87 Glioblastoma Cells

  • Haris, Khalilah (Department of Neurosciences, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Ismail, Samhani (Department of Neurosciences, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Idris, Zamzuri (Department of Neurosciences, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Abdullah, Jafri Malin (Department of Neurosciences, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Yusoff, Abdul Aziz Mohamed (Department of Neurosciences, School of Medical Sciences, Universiti Sains Malaysia)
  • Published : 2014.06.15

Abstract

Glioblastoma, the most aggressive and malignant form of glioma, appears to be resistant to various chemotherapeutic agents. Hence, approaches have been intensively investigated to targeti specific molecular pathways involved in glioblastoma development and progression. Aloe emodin is believed to modulate the expression of several genes in cancer cells. We aimed to understand the molecular mechanisms underlying the therapeutic effect of Aloe emodin on gene expression profiles in the human U87 glioblastoma cell line utilizing microarray technology. The gene expression analysis revealed that a total of 8,226 gene alterations out of 28,869 genes were detected after treatment with $58.6{\mu}g/ml$ for 24 hours. Out of this total, 34 genes demonstrated statistically significant change (p<0.05) ranging from 1.07 to 1.87 fold. The results revealed that 22 genes were up-regulated and 12 genes were down-regulated in response to Aloe emodin treatment. These genes were then grouped into several clusters based on their biological functions, revealing induction of expression of genes involved in apoptosis (programmed cell death) and tissue remodelling in U87 cells (p<0.01). Several genes with significant changes of the expression level e.g. SHARPIN, BCAP31, FIS1, RAC1 and TGM2 from the apoptotic cluster were confirmed by quantitative real-time PCR (qRT-PCR). These results could serve as guidance for further studies in order to discover molecular targets for the cancer therapy based on Aloe emodin treatment.

Acknowledgement

Supported by : Fundamental Research Grant Scheme (FRGS)

References

  1. Avecedo-Duncan M, Russel C, Patel S, Patel R (2004). Aloe-emodin modulates PKC isozymes, inhibits proliferation, and induces apoptosis in U-373MG glioma cells. Int Immunopharmacol, 4, 1775-84. https://doi.org/10.1016/j.intimp.2004.07.012
  2. Caffo M, Barresi V, Caruso G, et al (2013). Gliomas Biology: Angiogenesis and Invasion. In 'Evolution of the Molecular Biology of Brain Tumors and the Therapeutics Implications', Eds, Lichtor T. InTech Publisher, New York pp 37-103.
  3. Cha TL, Chuang MJ, Tang SH, et al (2013). Emodin modulates epigenetic modifications and suppresses bladder carcinoma cell growth. Mol Carcinog. [Epub ahead of print]
  4. Chen YY, Chiang SY, Lin JG, et al (2010). Emodin, aloe-emodin and rhein induced DNA damage and inhibited DNA repair gene expression in SCC-4 human tongue cancer cells. Anticancer Res, 30, 945-51.
  5. Cheng WC, Gong HY, Wan Y, Qiu YS, Wang Y (2012). In vitro risk evaluation of the inhibitory effects of Aloe emodin towards UDP-glucuronosyltransferases (UGTs). Lat Am J Pharm, 31, 1207-9.
  6. Chiu TH, Lai WW, Hsia TC, et al (2009). Aloe-emodin induces cell death through S-phase arrest and caspase-dependent pathways in human tongue squamous cancer SCC-4 cells. Anticancer Res, 29, 4503-11.
  7. Collins I, Workman P (2006). New approaches to molecular cancer therapeutics. Nat Chem Biol, 2, 689-700. https://doi.org/10.1038/nchembio840
  8. Espenshade PJ (2006). SREBPs: sterol-regulated transcription factors. J Cell Sci, 119, 973-6. https://doi.org/10.1242/jcs02866
  9. Goenka S, Kaplan MH (2011). Transcriptional regulation by STAT6. Immunol Res, 50, 87-96. https://doi.org/10.1007/s12026-011-8205-2
  10. Ettinger SL, Sobel R, Whitmore TG, et al (2004). Dysregulation of sterol response element-binding protein and downstream effectors in prostate cancer during progression to androgen independence. Cancer Res, 64, 2212-21. https://doi.org/10.1158/0008-5472.CAN-2148-2
  11. Fu J, Yang QY, Sai K, et al (2013). TGM2 inhibition attenuated ID1 expression in CD44 high glioma-initiating cells. Neuro-Oncology, 15, 1353-65. https://doi.org/10.1093/neuonc/not079
  12. Godoy PRDV, Mello SS, Donaires FS, et al (2013). In silico Analysis of Transcription Factors Associated to Differentially Expressed Genes in Irradiated Glioblastoma Cell Lines In 'Evolution of the Molecular Biology of Brain Tumors and the Therapeutics Implications', Eds, Lichtor T. InTech Publisher, New York pp 577-600.
  13. Gooch JL, Christy B, Yee D (2002). STAT6 mediates interleukin-4 growth inhibition in human breast cancer cells. Neoplasia, 4, 324-31. https://doi.org/10.1038/sj.neo.7900248
  14. He L, Bi JJ, Guo Q, et al (2012). Effects of emodin extracted from Chinese herbs on proliferation of non-small cell lung cancer and underlying mechanisms. Asian Pac J Cancer Prev, 13, 1505-10. https://doi.org/10.7314/APJCP.2012.13.4.1505
  15. Heemers H, Maes B, Foufelle F, et al (2001). Androgens stimulate lipogenic gene expression in prostate cancer cells by activation of the sterol regulatory element-binding protein cleavage activating protein/sterol regulatory element-binding protein pathway. Mol Endocrinol, 15, 1817-28. https://doi.org/10.1210/mend.15.10.0703
  16. Holmes RS (2012). Vertebrate patatin-like phospholipase domain-containing protein 4 (PNPLA4) genes and proteins: a gene with a role in retinol metabolism. 3 Biotechnology, 2, 277-86.
  17. Hu B, Shi B, Jarzynka MJ (2009). ADP-ribosylation factor 6 regulates glioma cell invasion through the IQ-domain GTPase-activating protein 1-Rac1-mediated pathway. Cancer Res, 69, 794-801. https://doi.org/10.1158/0008-5472.CAN-08-2110
  18. Iwasawa R, Mahul-Mellier AL, Datler C, Pazarentzos E, Grimm S (2011). Fis1 and Bap31 bridge the mitochondria-ER interface to establish a platform for apoptosis induction. EMBO J, 30, 556-68. https://doi.org/10.1038/emboj.2010.346
  19. Huang DW, Sherman BT, Lempicki RA (2009). Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc, 4, 44-57.
  20. Huang PH, Huang CH, Chen MC, et al (2013). Emodin and Aloe emodin suppress breast cancer cell proliferation through ERa inhibition. Evid Based Complement Alternat Med, 1-12.
  21. Igata T, Jinnin M, Makino T, et al (2010). Up-regulated type I collagen expression by the inhibition of Rac1 signaling pathway in human dermal fibroblasts. Biochem Biophys Res Commun, 393, 101-5. https://doi.org/10.1016/j.bbrc.2010.01.090
  22. Jang GY, Jeon JH, Cho SY, et al (2010). Transglutaminase 2 suppresses apoptosis by modulating caspase 3 and NF-kappaB activity in hypoxix tumor cells. Oncogene, 29, 356-67. https://doi.org/10.1038/onc.2009.342
  23. Jin W, Wu J, Zhunag Z, et al (2007). Gene expression profiling in apoptotic K562 treated by homoharringtonine. Acta Biochimica et Biophysica Sinica, 39, 982-91. https://doi.org/10.1111/j.1745-7270.2007.00364.x
  24. Jung J, Kim JM, Park B, et al (2010). Newly identified tumor-associated role of human Sharpin. Mol Cell Biochem, 340, 161-7. https://doi.org/10.1007/s11010-010-0413-x
  25. Kuo PL, Lin TC, Lin CC (2002). The antiproliferative activity of Aloe emodin is through p53-dependent and p-21 dependent apoptotic pathway in human hepatoma cell lines. Life Sci, 71, 1879-92. https://doi.org/10.1016/S0024-3205(02)01900-8
  26. Lee HZ (2001). Effects and mechanism of aloe-emodin on cell death in human lung squamous cell carcinoma. Eur J Pharmcol, 431, 287-95. https://doi.org/10.1016/S0014-2999(01)01467-4
  27. Lee YJ, Jeong SY, Karbowski CL, Youle, RJ (2004). Roles of mammalian mitochondrial fission mediators Fis1, Drp1 and Opa1 in apoptosis. Mol Biol Cell, 15, 5001-11. https://doi.org/10.1091/mbc.E04-04-0294
  28. LingYi D, KeWei J, YanBin Z, et al (2011). BAP31 is frequently overexpressed in patients with primary colorecteral cancer and correlates with better prognosis. Chinese Science Bulletin, 56, 2444-9. https://doi.org/10.1007/s11434-011-4610-0
  29. Levy DE, Darnell JE (2002). STATs: Transcriptional control and biological impact. Nav Rev Mol Cell Biol, 3, 651-62. https://doi.org/10.1038/nrm909
  30. Liang Y, Sundberg JP (2011). SHARPIN regulates mitochondria-dependent apoptosis in keratinocytes. J Dermatol Sci, 63, 148-53. https://doi.org/10.1016/j.jdermsci.2011.04.012
  31. Lim S, Sala C, Yoon J, et al (2001). Sharpin, a novel postsynaptic density protein that directly interacts with the shank family of proteins. Mol Cell Neurosci, 17, 385-97. https://doi.org/10.1006/mcne.2000.0940
  32. Lu GD, Shen HM, Ong CN, Chung MC (2007). Anticancer effects of aloe-emodin on HepG2 cells: Cellular and proteomic studies. Proteomics Clin Appl, 1, 410-9. https://doi.org/10.1002/prca.200600798
  33. Mai SKM, Auburger G, Bereiter-Hahn J, Jendrach M (2010). Decreased expression of Drp1 anf Fis1 mediates mitochondrial elongation in senescent cells and enhances resistance to oxidative stress through PINK1. J Cell Sci, 123, 917-26. https://doi.org/10.1242/jcs.059246
  34. Mandal C, Chandra S, Schauer R (2012). Regulation of O-acetylation of sialic acids by sialate-O-acetyltransferase and sialate-O-acetylesterase activities in childhood acute lymphoblastic leukemia. Glycobiology, 22, 70-83. https://doi.org/10.1093/glycob/cwr106
  35. Marin-Valencia I, Good LB, Ma Q, Malloy CR, Pascual JM (2013). Heptanoate as a neural fuel: energetic and neurotransmitter precursors in normal and glucose transporter I-deficient (G1D) brain. J Cereb Blood Flow Metab, 33, 175-82. https://doi.org/10.1038/jcbfm.2012.151
  36. McGill GG, Horstmann M, Widlund HR, et al (2002). Bcl2 regulation by the melanocyte master regulator mitf modulates lineage survival and melanoma cell viability. Cell, 109, 707-18. https://doi.org/10.1016/S0092-8674(02)00762-6
  37. Pecere T, Gazzola MT, Mucignat C, et al (2000). Aloe-emodin is a new type of anticancer agent with selective activity against neuroectodermal tumors. Cancer Res, 60, 2800-4.
  38. Mehta K, Fok J, Miller FR, Koul D, Sahin AA (2004). Prognostic significance of tissue transglutaminase in drug resistant and metastatic breast cancer. Clin Cancer Res, 10, 8068-76. https://doi.org/10.1158/1078-0432.CCR-04-1107
  39. Milakovic T, Tucholski J, McCoy E, Johnson GV (2004). Intracellular localization and activity state of tissue transglutaminase differentially impacts cell death. J Biol Chem, 279, 8715-22. https://doi.org/10.1074/jbc.M308479200
  40. Moniz S, Martinho O, Pinto F, et al (2013). Loss of WNK2 expression by promoter gene methylation occurs in adult gliomas and triggers Rac1-mediated tumour cell invasiveness. Hum Mol Genet, 22, 84-95. https://doi.org/10.1093/hmg/dds405
  41. Pfaffl MW, Horgan GW, Dempfle L (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res, 29, 2002-7.
  42. Qin HB, Zhu M, Du TK, et al (2010). Effects of aloe-emodin on proliferation and migration of human gastric cancer cell line BGC-823. Acta Anatomica Sinica, 41, 909-11.
  43. Rich JN, Hans C, Jones B, et al (2005). Gene expression profiling and genetic markers in glioblastoma survival. Cancer Res, 65, 4051-58. https://doi.org/10.1158/0008-5472.CAN-04-3936
  44. Sharma SM, Bronisz A, Hu R, et al (2007). MITF and PU.1 recruit p38 MAPK and NFATc1 to target genes during osteoclast differentiation. J Biol Chem, 282, 15921-29. https://doi.org/10.1074/jbc.M609723200
  45. Suboj P, Babykutty S, Srinivas P, Gopala S (2012) (I). Aloe emodin induces G2/M cell cycle arrest and apoptosis via activation of caspase-6 in human colon cancer cells. Pharmacology, 89, 91-8. https://doi.org/10.1159/000335659
  46. Tachibana M, Takeda K, Nobukuni Y, et al (1996). Ectopic expression of MITF, a gene for Waardenburg syndrome type 2, converts fibroblasts to cells with melanocyte characteristics. Nat Genet, 14, 50-4. https://doi.org/10.1038/ng0996-50
  47. Suboj P, Babykutty S, Valiyaparambil Gopi DR, et al (2012) (II). Aloe emodin inhibits colon cancer cell migration/angiogenesis by downregulating MMP-2/9, RhoB and VEGF via reduced DNA binding activity of NF-${\kappa}B$. Eur J Pharm Sci, 45, 581-91. https://doi.org/10.1016/j.ejps.2011.12.012
  48. Sun D, Xu D, Zhang B (2006). Rac signaling in tumorigenesis and as target for anticancer drug development. Drug Resist Update, 9, 274-87. https://doi.org/10.1016/j.drup.2006.12.001
  49. Tabolacci C, Oliverio S, Lentini A, et al (2011). Aloe-emodin as antiproliferative and differentiating agent on human U937 monoblastic leukemia cells. Life Sci, 89, 812-20. https://doi.org/10.1016/j.lfs.2011.09.008
  50. Tavian D, Missaglia S, Redaelli C, et al (2012). Contribution of novel ATGL missense mutations to the clinical phenotype of NLSD-M: a strikingly low amount of lipase activity may preserve cardiac function. Hum Mol Genet, 21, 5318-28. https://doi.org/10.1093/hmg/dds388
  51. Wakana Y, Takai S, Nakajima K, et al (2008). Bap31 is an itirent protein that moves between the peripheral endoplasmic reticulum (ER) and a juxtanuclear compartment related to ER-associated degradation. Mol Cell Biol, 19, 1825-36. https://doi.org/10.1091/mbc.E07-08-0781
  52. Wasserman L, Avigad S, Beery E, Nordenberg J, Fening E (2002). The effect of Aloe emodin on the proliferation of new merkel carcinoma cell line. Am J Dermatopathol, 24, 17-22. https://doi.org/10.1097/00000372-200202000-00003
  53. Williams KJ, Argus JP, Zhu Y, et al (2013). An essential requirement for the SCAP/SREBP signaling axis to protect cancer cells from lipotoxicity. Cancer Res, 73, 2850-62. https://doi.org/10.1158/0008-5472.CAN-13-0382-T
  54. Zhao RZ (2012). Targeting Effect of Traditional Chinese Medicine. In 'Recent Advances in Theories and Practice of Chinese Medicine', Eds Kuang H. InTech Publisher, New York pp 313-36.
  55. Wintner LM, Giesinger JM, Holzner B (2011). Patient-Reported Outcome Monitoring in Brain Tumour Patients: Benefits and Requirements. In 'Diagnostics Tehniques and Surgical Management of Brain Tumors' Eds, Abujamra AL. InTech Publisher, New York pp 3-26.
  56. Yoshida T, Zhang Y, Leslie A, et al (2010). Blockade of Rac1 activity induces G1 cell cycle arrest or apoptosis in breast cancer cells through downregulation of Cyclin D1, Survivin and X-linked inhibitor of apoptosis protein. Mol Cancer Ther, 9, 1657-68. https://doi.org/10.1158/1535-7163.MCT-09-0906
  57. Zhang Y, Chan DC (2007). Structural basis for recruitment of mitochondrial fission complexes by Fis1. Proc Natl Acad Sci USA, 104, 18526-30. https://doi.org/10.1073/pnas.0706441104

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