Please use this identifier to cite or link to this item:
標題: Gefitinib誘發神經膠細胞瘤細胞凋亡機制探討
Molecular mechanisms of gefitinib-induced apoptosis in experimental glioma cells
作者: 張正一
Chang, Cheng-Yi
關鍵字: Gefitinib;Gefitinib;glioma cell;apoptosis;神經膠細胞瘤;細胞凋亡
出版社: 生命科學系所
引用: 1. Louis, D.N., Ohgaki, H., Wiestler, O.D.(2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 113(2): 97-109 2. Nagane M, Huang HJ, Cavenee WK (1997) Advances in the molecular genetics of gliomas. Curr Opin Oncol 9:215-222 3. Soni D, King JA, Kaye AH, Hovens CM (2005) Genetics of glioblastoma multiforme: mitogenic signaling and cell cycle pathway converge. J Clin Neurosci 12:1-5 4. Mendelsohn J, Baselga J (2000) The EGF receptor family as targets for cancer therapy. Oncogene, 56:6550-6565. 5. Frederick L, Wang XY, Eley G (2000) James CD: Diversity and frequency of epidermal growth factor receptor mutations in human glioblastomas. Cancer Res, 60:1383-1387. 6. Shinojima N, Tada K, Shiraishi S (2003) Prognostic value of epidermal growth factor receptor in patients with glioblastoma multiforme. Cancer Res ,63:6962-6970 7. Citri A, Yarden Y.(2006) EGF-ERBB signalling: towards the systems level. Nat Rev Mol Cell Biol ;7: 505-16. 8. Hynes NH, Lane HA (2005). ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer ;5:341-54 9. Ciardiello F. Tortora G (2008). EGFR Antagonists in Cancer Treatment (Review) N Engl J Med 358;11, 1160-1174 10. Aldape KD, Ballman K, Furth A(2004) Immunohistochemical detection of EGFRvIII in high malignancy grade astrocytomas and evaluation of prognostic significance. J Neuropathol Exp Neurol ;63:700-7 11. Sugawa N, Ekstrand AJ, James CD (1990). Identical splicing of aberrant epidermal growth factor receptor transcripts from amplified rearranged genes in human glioblastomas. Proc Natl Acad Sci U S A 87:8602-6. 12. Kim ES, Khuri FR, Herbst RS. (2001)Epidermal growth factor receptor biology (IMC-C225). Curr Opin Oncol 13: 506-513 13. Foltz I, King C, Liang M . Panitumumab induces internalization of the epidermal growth facor (EGFr). Presented at: AAACR-NCI-EORTC International Conference on Molecular Target and Cancer Therapeutics; November 14-18 , 2005; Pholadelphia, PA. Abstract B43 14. Jiang Z, Zheng X, Rich KM. (2003) Down-regulation of Bcl-2 and Bcl-xL expression with bispecific antisense treatment in glioblastoma cell lines induce cell death. J Neurochem. Jan; 84(2):273-81. 15. Ichinose M, Liu XH, Hagihara N (2002) Extracellular Bad fused to toxin transport domains induces apoptosis. Cancer Res. 1;62(5):1433-8. 16. Cartron PF, Oliver L, Martin S (2002) The expression of a new variant of the pro-apoptotic molecule Bax, Baxpsi, is correlated with an increased survival of glioblastoma multiforme patients. Hum Mol Genet. 15;11(6):675-87 17. Busser B, Sancey L, Josserand V (2010) Amphiregulin promotes BAX inhibition and resistance to gefitinib in non-small-cell lung cancers. Mol Ther.;18(3):528-35. 18. Turkson J, Bowman T, Garcia Chao DT.(1998) BCL-2 family: regulators of cell death. Annu Rev Immunol 16:395-419 19. Gleichmann M, Weller M, Schulz JB, (2000) Insulin-like growth factor-1-mediated protection from neuronal apoptosis is linked to phosphorylation of the pro-apoptotic protein BAD but not to inhibition of cytochrome c translocation in rat cerebellar neurons. Neurosci Lett, 282(1-2):69-72 20. Green DR, Reed JC (1998), Mitochondria and apoptosis. Science 281(5381):1309-1312 21. Thornberry NA, Lazebnik Y,(1998) Caspases: enemies within. Science, 281(5381):1312-1316 22. Riese II, D.J., Stern, D.F., (1998). Specificity within the EGF family/ErbB receptor family signaling network. Bioessays 20, 41-48. 23. Dent, P., Reardon, D.B., Park, J.S.( 1999). Radiation-induced release of transforming growth factor alpha activates the epidermal growth factor receptor and mitogen-activated protein kinase pathway in carcinoma cells, leading to increased proliferation and protection from radiation-induced cell death. Mol. Biol. Cell 10, 2493-2506 24. Dong, J., Wiley, H.S, (2000). Trafficking and proteolytic release of epidermal growth factor receptor ligands are modulated by their membraneanchoring domains. J. Biol. Chem. 275, 557-564 25. Roudabush, F.L., Pierce, K.L., Maudsley, S (2000). Transactivation of the EGF receptor mediates IGF-1-stimulated shc phosphorylation and ERK1/2 activation in COS-7 cells. J. Biol.Chem. 275, 22583-22589. 26. Wakeling, A.E., Guy, S.P., Woodburn, J.R., (2002). ZD1839 (Iressa): an orally active inhibitor of epidermal growth factor signaling with potential for cancer therapy. Cancer Res. 62, 5749-5754. 27. Wikstrand, C.J., Reist, C.J., Archer, G.E., Zalutsky, M.R., Bigner, D.D.,(1998). The class III variant of the epidermal growth factor receptor (EGFRvIII): characterization and utilization as an immunotherapeutic target. J. Neurovirol. 4, 148-158 28. Learn, C.A., Hartzell, T.L., Wikstrand, C.J., (2004). Resistance to tyrosine kinase inhibition by mutant epidermal growth factor receptor variant III contributes to the neoplastic phenotype of glioblastoma multiforme. Clin. Cancer Res. 10, 3216-3224. 29. Li, B., Chang, C.M., Yuan, M. (2003). Resistance to small molecule inhibitors of epidermal growth factor receptor in malignant gliomas. Cancer Res. 63, 7443-7450. 30. Li, B., Yuan, M., Kim, I.A.,(2004). Mutant epidermal growth factor receptor displays increased signaling through the phosphatidylinositol-3 kinase/AKT pathway and promotes radioresistance in cells of astrocytic origin. Oncogene 23, 4594-4602. 31. Janmaat, M.L., Kruyt, F.A., Rodriguez, J.A., (2003). Response to epidermal growth factor receptor inhibitors in nonsmall cell lung cancer cells: limited antiproliferative effects and absence of apoptosis associated with persistent activity of extracellular signal-regulated kinase or Akt kinase pathways. Clin. Cancer Res. 9, 2316-2326. 32. Wymann, M.P., Pirola, L., (1998). Structure and function of phosphoinositide 3-kinases. Biochim. Biophys. Acta 1436, 127-150. 33. Kolch, W., Calder, M., Gilbert, D.,( 2005). When kinases meet mathematics: the systems biology of MAPK signalling. FEBS Lett. 579, 1891-1895. 34. Gilmore, A.P., Valentijn, A.J., Wang, P.( 2002). Activation of BAD by therapeutic inhibition of epidermal growth factor receptor and transactivation by insulin-like growth factor receptor. J. Biol. Chem. 277, 27643-27650. 35. Yakes, F.M., Chinratanalab, W., Ritter, C.A.( 2002). Herceptin-induced inhibition of phosphatidylinositol-3 kinase and Akt Is required for antibody-mediated effects on p27, cyclin D1, and antitumor action. Cancer Res. 62, 4132-4141 36. Jones, H.E., Goddard, L., Gee, J.M.(2004). Insulin-like growth factor-I receptor signalling and acquired resistance to gefitinib (ZD1839; Iressa) in human breast and prostate cancer cells. Endocr. Relat. Cancer 11, 793-814 37. Chakravarti, A., Loeffler, J.S., Dyson, N.J., (2002). Insulin-like growth factor receptor I mediates resistance to anti-epidermal growth factor receptor therapy in primary human glioblastoma cells through continued activation of phosphoinositide 3-kinase signaling. Cancer Res. 62, 200-207 38. Stegmaier K, Corsello SM, Ross KN, (2005) Gefitinib induces myeloid differentiation of acute myeloid leukemia. Blood. 15;106(8):2841-8 39. Okubo S, Kurebayashi J, Otsuki T(2004) Additive antitumour effect of the epidermal growth factor receptor tyrosine kinase inhibitor gefitinib (Iressa,ZD1839) and the antioestrogen fulvestrant in breast cancer cells. Br J Cancer 90:236-244, 2004 40. Rich JN, Reardon DA, Peery T, Dowell JM, Quinn JA, Penne KL, Wikstrand CJ, Van Duyn LB, Dancey JE, McLendon RE, Kao JC, Stenzel TT, Ahmed Rasheed BK, Tourt-Uhlig SE, Herndon JE 2nd, Vredenburgh JJ, Sampson JH, Friedman AH, Bigner DD, Friedman HS (2004) Phase II trial of gefitinib in recurrent glioblastoma. J Clin Oncol 22:133-142 41. Guillamo JS, de Boüard S, Valable S (2009). Molecular Mechanisms Underlying Effects of Epidermal Growth Factor Receptor Inhibition on Invasion, Proliferation, and Angiogenesis in Experimental Glioma. Clin Cancer Res 2009; 15 (11) June 1, (Published Online ) 42. Chang GC, Hsu SL, Tsai JR, Liang FP, Lin SY, Sheu GT, Chen CY (2004) Molecular mechanisms of ZD1839-induced G1-cell cycle arrest and apoptosis in human lung adenocarcinoma A549 cells. Biochem Pharmacol 68:1453-1464 43. Chang GC, Yu CT, Tsai CH, Tsai JR, Chen JC, Wu CC, Wu WJ, Hsu SL (2008) An epidermal growth factor inhibitor, gefitinib, induces apoptosis through a p53-dependent upregulation of pro-apoptotic molecules and downregulation of anti-apoptotic molecules in human lung adenocarcinoma A549 cells. Eur J Pharmacol 600:37-44 44. Piechocki MP, Yoo GH, Dibbley SK, Amjad EH, Lonardo F (2006) Iressa induces cytostasis and augments Fas-mediated apoptosis in acinic cell adenocarcinoma overexpressing HER2/neu. Int J Cancer 119:441-454 45. Rho JK, Choi YJ, Ryoo BY, Na II, Yang SH, Kim CH, Lee JC (2007) p53 enhances gefitinib-induced growth inhibition and apoptosis by regulation of Fas in non-small cell lung cancer. Cancer Res 67:1163-1169 46. Ariyama H, Qin B, Baba E, Tanaka R, Mitsugi K, Harada M, Nakano S (2006) Gefitinib, a selective EGFR tyrosine kinase inhibitor, induces apoptosis through activation of Bax in human gallbladder adenocarcinoma cells. J Cell Biochem 97:724-734 47. Cragg MS, Kuroda J, Puthalakath H, Huang DCS, Strasser A (2007) Gefitinib-induced killing of NSCLC cell lines expressing mutant EGFR requires BIM and can be enhanced by BH3 mimetics. PLoS Med 4:e316 48. Sun Q, Ming L, Thomas SM, Wang Y, Chen ZG, Ferris RL, Grandis JR, Zhang L, Yu J. (2009) PUMA mediates EGFR tyrosine kinase inhibitor-induced apoptosis in head and neck cancer cells. Oncogene 28:2348-2357 49. Youle RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47-59 50. Kroemer G, Galluzzi L, Brenner C (2007) Mitochondrial membrane permeabilization in cell death. Physiol Rev 87:99-163 51. Harada H, Becknell B, Wilm M, Mann M, Huang LJ, Taylor SS, Scott JD, Korsmeyer SJ (1999) Phosphorylation and inactivation of BAD by mitochondria-anchored protein kinase A. Mol Cell 3:413-422 52. Danial NN (2009) BAD: undertaker by night, candyman by day. Oncogene 27:553-570 53. Zha J, Harada H, Yang E, Jocker J, Korsmeyer SJ (1996) Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell 87:619-628 54. Chiang CW, Kanies C, Kim KW, Fang WB, Parkhurst C, Xie M, Henry T, Yang E. (2003) Protein phosphatase 2A dephosphorylation of phosphoserine 112 plays the gatekeeper role for BAD-mediated apoptosis. Mol Cell Biol 23:6350-6362 55. Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91:231-241 56. Shimamura A, Ballif BA, Richards SA, Blenis J (2000) Rsk1 mediates a MEK-MAP kinase cell survival signal. Curr Biol 10:127-135 57. Tortora G, Caputo R, Damiano V, Fontanini G, Melisi D, Veneziani BM, Zunino F, Bianco AR, Ciardiello F (2001) Oral administration of a novel taxane, an antisense oligonucleotide targeting protein kinase A, and the epidermal growth factor receptor inhibitor Iressa causes cooperative antitumor and antiangiogenesis activity. Clin Cancer Res 7:4156-4163 58. Cohen P, Holmes CFB, Tsukitani Y (1990) Okadaic acid: a new probe for the study of cellular regulation. Trends Biochem Sci 15:98-102 59. Xin M, Deng X (2006) Protein phosphatase 2A enhances the proapoptotic function of Bax through dephosphorylation. J Biol Chem 281:18859-18867 60. Garibal J, Hollville É, Renouf B, Tétaud C, Wiels J (2010) Caspase-8-mediated cleavage of Bid and protein phosphatase 2A-mediated activation of Bax are necessary for verotoxin-1-induced apoptosis in Burkitt's lymphoma cells. Cell Signaling 22:467-475 61. Mellinghoff IK, Wang MY, Vivanco I, Haas-Kogan DA, Zhu S, Dia EQ, Lu KV, Yoshimoto K, Huang JH, Chute DJ, Riggs BL, Horvath S, Liau LM, Cavenee WK, Rao PN, Beroukhim R, Peck TC, Lee JC, Sellers WR, Stokoe D, Prados M, Cloughesy TF, Sawyers CL, Mischel PS (2005). Molecular determinants of the response of glioblastomas to EGFR kinase inhibitors. N Engl J Med 353:2012-2024 62. Krex D, Mohr B, Hauses M, Ehninger G, Schackert HK, Schackert G (2001) Identification of uncommon chromosomal aberrations in the neuroglioma cell line H4 by spectral karyotyping. J Neurooncol 52:119-128 63. Rubenstein M, Shaw M, Mirochnik Y, Slobodskoy L, Glick R, Lichtor T, Chou P, Guinan P (1999) In vivo establishment of T98G human glioblastoma. Methods Find Exp Clin Pharmacol 21:391-393 64. Zhang R, Banik NL, Ray SK (2007) Combination of all-trans retinoic acid and interferon-gamma suppressed PI3K/Akt survival pathway in glioblastoma T98G cells whereas NF-kB survival signaling in glioblastoma U87MG cells for induction of apoptosis. Neurochem Res 32:2194-2202 65. Lal B, Goodwin CR, Sang Y, Foss CA, Cornet K, Muzamil S, Pomper MG, Kim J, Laterra J (2009) EGFRvIII and c-Met pathway inhibitors synergize against PTEN-null/EGFRvIII+ glioblastoma xenografts. Mol Cancer Ther 8:1751-1760 66. Premkumar DR, Arnold B, Pollack IF (2006) Cooperative inhibitory effect of ZD1839 (Iressa) in combination with 17-AAG on glioma cell growth. Mol Carcinogenesis 45:288-301 67. Tamura S, Hosoi H, Kuwahara Y, Kikuchi K, Otabe O, Izumi M, Tsuchiya K, Iehara T, Gotoh T, Sugimoto T (2007) Induction of apoptosis by an inhibitor of EGFR in neuroblastoma cells. Biochem Biophys Res Commun 358:226-232 68. Nakagawa K, Tamura T, Negoro S, Kudoh S, Yamamoto N, Yamamoto N, Takeda K, Swaisland H, Nakatani I, Hirose M, Dong RP, Fukuoka M (2003) Phase I pharmacokinetic trial of the inhibitor gefitinib (Iressa, ZD1839) in Japanese patients with solid malignant tumors. Ann Oncol 14:922-930 69. Cemeus C, Zhao TT, Barrett GM, Lorimer IA, Dimitroulakos J (2008) Lovastatin enhances gefitinib activity in glioblastoma cells irrespective of EGFRvIII and PTEN status. J Neurooncol 90:9-17 70. Doherty L, Gigas DC, Kesari S, Drappatz J, Kim R, Zimmerman J, Ostrowsky L, Wen PY (2006) Pilot study of the combination of EGFR and mTOR inhibitors in recurrent malignant gliomas. Neurology 67:156-158 71. Geoerger B, Gaspar N, Opolon P, Morizet J, Devanz P, Lecluse Y, Valent A, Lacroix L, Grill J, Vassal G (2008) EGFR tyrosine kinase inhibition radiosensitizes and induces apoptosis in malignant glioma and childhood ependymoma xenografts. Int J Cancer 123:209-216 72. Gustafson DL, Frederick B, Merz AL, Raben D (2008) Dose scheduling of the dual VEGFR and EGFR tyrosine kinase inhibitor vandetanib (ZD6474, Zactima) in combination with radiotherapy in EGFR-positive and EGFR-null human head and neck tumor xenografts. Cancer Chemother Pharmacol 61:179-188 73. Hegi ME, Diserens AC, Bady P, Kamoshima Y, Kouwenhoven MCM, Delorenzi M, Lambiv WL, Hamou MF, Matter MS, Koch A, Heppner FL, Yonekawa Y, Merlo A, Frei K, Mariani L, Hofer S (2011) Pathway analysis of glioblastoma tissue after preoperative treatment with the EGFR tyrosine kinase inhibitor gefitinib - a phase II trial. Mol Cancer Ther DOI: 10.1158/1535-7163. MCT-11-0048 74. Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science. 2002 Dec 6;298(5600):1911-2 75. Teiji Wada and Josef M Penninger Mitogen-activated protein kinases in apoptosis regulation Oncogene (2004) 23, 2838-2849 76. Kenji Takeuchi, Tomohiro Shin-ya, Kazuto Nishio and Fumiaki Ito Mitogen-activated protein kinase phosphatase-1 modulated JNK activation is critical for apoptosis induced by inhibitor of epidermal growth factor receptor-tyrosine kinase FEBS Journal 276 (2009) 1255-1265 77. Choe G, Horvath S, Cloughesy TF, Crosby K, Seligson D, Palotie A, Inge L, Smith BL, Sawyers CL, Mischel PS..Analysis of the phosphatidylinositol 3''-kinase signaling pathway in glioblastoma patients in vivo. Cancer Res 2003;63:2742-6. 78. Chen TC, Hinton DR, Zidovetzki R, Hofman FM: Up-regulation of the cAMP/PKA pathway inhibits proliferation, induces differentiation, and leads to apoptosis in malignant gliomas. Lab Invest 1998; 78: 165-174. 79. Helmbrecht K, Rensing L: Different constitutive heat shock protein 70 expression during proliferation and differentiation of rat C6 glioma cells. Neurochem Res 1999; 24: 1293-1299. 80. Farias CB, Lima RC, Lima LO, Flores DG, Meurer L, Brunetto AL, Schwartsmann G, Roesler R. Stimulation of proliferation of U138-MG glioblastoma cells by gastrin-releasing peptide in combination with agents that enhance cAMP signaling Oncology 2008;75:27-31
Gefitinib 是一種選擇性作用於上皮生長因子接受器的酪氨酸激酶抑制劑,目前仍嘗試於臨床上治療癌症病患,包括神經膠細胞瘤患者。然而目前對Gefitinib於神經膠細胞瘤所產生抑制癌細胞的詳細分子作用機轉仍不甚明瞭。Gefitinib可抑制神經膠細胞的生長,並誘使細胞產生細胞凋亡。於實驗中可發現Gefitinib經由內在細胞凋亡路徑誘使H4細胞死亡,其中作用的過程包括Bax蛋白於粒線體的位置改變,粒線體外膜的通透性改變,細胞色素C釋放至細胞質,caspase-3及caspase-9的活化。Gefitinib造成的神經膠細胞瘤凋亡的現象,可經由加入Bax siRNA及Bax蛋白通道抑制劑的方式抑制Bax的表現來緩和細胞凋亡,此現象說明Bax在此細胞凋亡過程扮演重要角色。Gefitinib造成Bad蛋白去磷酸化,特別是在Ser-112的位置上,此過程使Bad蛋白與Bcl-2及Bcl-xL結合的狀況增加。而Gefitinib造成Bad蛋白去磷酸化會伴隨著cAMP含量減少及protein kinase A的活性降低。Adenylyl cyclase 活化劑forskolin 可減緩Gefitinib造成的Bad蛋白去磷酸化,Bax蛋白於粒線體膜上位置的改變,caspase-3及caspase-9的活化,及細胞活性喪失等現象,而蛋白激酶A抑制劑H89則具有與forskolin相反的效果。而較無法解釋的是加入非選擇性蛋白磷酸酶抑制劑okadaic acid,可減緩Gefitinib所造成的影響,卻不改變Bad蛋白去磷酸化的程度。類似的情形在不同細胞株U87及T98G亦能觀察到,而U87本身具有較高的蛋白激酶A的活性。相對於H4細胞,U87對Gefitinib的反應較為遲緩且抗藥性較高。降低蛋白激酶A的活性可使H4,T98G及U87對Gefitinib毒性的敏感度增加,Bad於Serine-112位置去磷酸化程度,及caspase-3/caspase-9活化程度增加。本實驗發現gefitinib造成的神經膠細胞瘤凋亡的過程與Bad/Bax蛋白訊息傳導路徑有關。蛋白激酶A去活化在促使Bad蛋白引發細胞凋亡的過程中扮演著一定角色。

Gefitinib, a selectively epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, is under clinical test and use in cancer patients, including glioma. However, the molecular mechanisms involved in gefitinib-mediated anticancer effects against glioma remain largely uncharacterized. Gefitinib inhibited cell growth and induced apoptosis in human glioma cells. Gefitinib induced death of H4 cells with characteristics of intrinsic apoptotic pathway, including Bax mitochondrial translocation, mitochondrial outer membrane permeabilization, cytochrome c cytosolic release, and caspase-9/caspase-3 activation. The importance of Bax in mediating gefitinib-induced apoptosis was confirmed by the attenuation of apoptosis by Bax siRNA and Bax channel blocker. Gefitinib caused Bad dephosphorylation, particularly in serine-112, and increased its binding preference to Bcl-2 and Bcl-xL. The dephosphorylation of Bad in gefitinib-treated cells was accompanied by decreased intracellular cyclic AMP content and protein kinase A (PKA) activity. Adenylyl cyclase activator forskolin attenuated, but PKA inhibitor H89 augmented, gefitinib-induced Bad dephosphorylation, Bax mitochondrial translocation, caspase-9/caspase-3 activation, and viability loss. Intriguingly, a nonselective protein phosphatase inhibitor okadaic acid alleviated gefitinib-induced alterations, except Bad dephosphorylation. In parallel with the higher basal PKA activity, U87 cells showed a delayed and relatively resistant response to gefitinib treatment than H4 and T98G cells. The inactivation of PKA sensitized H4, T98G, and U87 cells towards gefitinib cytotoxicity, Bad dephosphorylation in serine-112, and caspase-9/caspase-3 activation. Our findings suggest the involvement of the Bad/Bax signaling pathway in gefitinib-induced glioma apoptosis. Furthermore, the inactivation of PKA was shown to play a role in triggering the proapoptotic function of Bad.
其他識別: U0005-2007201110320100
Appears in Collections:生命科學系所

Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.