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系統識別號 U0026-2807201415251200
論文名稱(中文) 新穎α-咔吧啉衍生物TJY-16誘導惡性神經膠質瘤死亡機轉之探討
論文名稱(英文) Induction of Malignant Glioma Cell Death by a Novel Synthesized α-carboline Derivative TJY-16
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 102
學期 2
出版年 103
研究生(中文) 黃筱杰
研究生(英文) Hsiao-Chieh Huang
學號 S26014065
學位類別 碩士
語文別 英文
論文頁數 50頁
口試委員 指導教授-簡伯武
口試委員-陳柏熹
口試委員-張雋曦
中文關鍵字 惡性神經膠質瘤  咔吧啉衍生物  細胞凋亡 
英文關鍵字 malignant glioma  carboline derivatives  apoptosis 
學科別分類
中文摘要 惡性神經膠質瘤為中樞神經系統中最致命且好發於成人的原發性腦瘤,這些腫瘤會無法控制地增生和侵略正常組織,儘管發生率比肺癌低,但是病患經由手術切除、放射線或化學治療,預後仍不佳且復發率相當高,平均存活率約14.6個月,因此研發新療法仍是迫切而重要的。α-咔吧啉衍生物主要存在於海洋動物,先前已研究出此類化合物具有抗瘧原蟲、抗病毒甚至抗癌的作用,因此本研究將探討以化學合成方式合成出的新穎α-咔吧啉衍生物TJY,是否具有治療惡性神經膠質瘤之能力,並選出最有效的TJY,再進一步探討其中相關的機制。首先,我們篩選6個TJY藥物對3株人類神經膠質瘤細胞株U87、U251、T98G及1株大白鼠神經膠質瘤C6的抗癌作用,WST-1 assay 結果顯示TJY-16具有最強抑制細胞存活率的能力。進一步再利用trypan blue exclusion assay觀察細胞生長情形及死亡率,發現TJY-16對於三株神經膠質瘤細胞具有生長抑制作用,且可增加細胞的死亡率。而經由流式細胞儀分析的結果顯示,在給予TJY-16後,神經膠質瘤細胞會產生G2/M phase停滯的現象,且伴隨著sub-G1的增加。此外更利用Hoechst 33342染色發現TJY-16也會造成細胞核濃染現象的產生。西方墨點法則顯示cleaved caspase-3、cleaved caspase-8表現量明顯增加,再加上利用流式細胞儀分析得知粒線體膜電位會下降。綜合以上結果,我們推測TJY-16會導致神經膠質瘤細胞走向細胞凋亡的途徑。而在皮下動物模式中,也證實了TJY-16可以抑制神經膠質瘤生長,且不影響動物的體重以及其器官。這些結果皆顯示了TJY-16可以有效的抑制神經膠質瘤,希望在未來可以為治療腦瘤提供一個新的契機。
英文摘要 Malignant gliomas are the most lethal primary brain tumors of the central nervous system. Although the incidence of brain tumor is lower than lung cancer, brain tumor is rarely curable in spite of receiving surgery, chemotherapy and radiotherapy. The median survival is only 14.6 months. Therefore, it is important and urgently needed to develop a potential anti-malignant glioma drug. The α-carboline (pyrido[2,3-b]indole) derivatives were abundant in the marine animals. Previous studies have focused on their biological activities such as anti-plasmodial, anti-virus and anti-tumor. We aim to investigate the anti-tumor activity and mechanisms of novel synthesized α-carboline derivatives TJY against malignant gliomas. After screening 6 TJY compounds, we found that TJY-16 (6-acetyl-9- (3,4,5-trimethoxybenzyl)-9H-pyrido[2,3-b]indole) had the most powerful anti-glioma effects. TJY-16 decreased cell viability of C6, U87, T98G and U251 glioma cell lines in a dose- and time-dependent manner without affecting normal glia cell line SVGP12. Trypan blue exclusion assay further showed the reduction of cell viability was due to both cell growth inhibition and cell death induction after TJY-16 treatment in U87, T98G and U251 glioma cells. Flow cytometric analysis indicated that both G2/M phase and sub-G1 phase increased after treatment with TJY-16. Using Hoechst staining of nucleus, we found that TJY-16 induced chromatin condensation and DNA fragmentation, typical features of apoptosis. Western blotting showed the activation of caspase-8, a crucial upstream mediator of extrinsic apoptotic pathway. Furthermore, JC-1 staining demonstrated that depolarization of mitochondrial membrane potential (ΔΨm) occurred in cells treated with TJY-16 in a time-dependent manner. It suggested that both extrinsic and intrinsic apoptotic pathway were involved in TJY-16-induced apoptosis. Finally, the subcutaneous xenograft model showed that the growth rate of tumor in TJY-16-treated group was slower than control group. The body weight of nude mice and H&E staining for organ histology revealed that there was no obvious difference between control and TJY-16-treated group. It indicated that TJY-16 effectively inhibited glioma in vitro and in vivo. In summary, we suggested that TJY-16 seemed to be a promising agent in the treatment of malignant gliomas.
論文目次 中文摘要(Abstract in Chinese) I
英文摘要(Abstract in English) III
誌謝 VI
目錄(Contents) VII
圖表索引(List of Figures and Tables) VIII
縮寫檢索表(Abbreviations) X
Chapter 1 Introduction 1
Chapter 2 Specific aims 13
Chapter 3 Materials and Methods 15
Chapter 4 Results 22
Chapter 5 Discussion 28
References 33
參考文獻 Abbott, N.J., Ronnback, L., and Hansson, E. (2006). Astrocyte-endothelial interactions at the blood-brain barrier. Nature reviews Neuroscience 7, 41-53.

Abe, A., Yamada, H., Moriya, S., and Miyazawa, K. (2011). The beta-carboline alkaloid harmol induces cell death via autophagy but not apoptosis in human non-small cell lung cancer A549 cells. Biological & pharmaceutical bulletin 34, 1264-1272.

Aguirre-Ghiso, J.A. (2007). Models, mechanisms and clinical evidence for cancer dormancy. Nature reviews Cancer 7, 834-846.

Ashok, P., Ganguly, S., and Murugesan, S. (2013). Review on in-vitro anti-malarial activity of natural beta-carboline alkaloids. Mini reviews in medicinal chemistry 13, 1778-1791.

Brandes, A.A., Tosoni, A., Franceschi, E., Sotti, G., Frezza, G., Amista, P., Morandi, L., Spagnolli, F., and Ermani, M. (2009). Recurrence pattern after temozolomide concomitant with and adjuvant to radiotherapy in newly diagnosed patients with glioblastoma: correlation With MGMT promoter methylation status. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 27, 1275-1279.

Cancer Genome Atlas Research Network (2008). Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455, 1061-1068.

Castedo, M., Perfettini, J.L., Roumier, T., Andreau, K., Medema, R., and Kroemer, G. (2004). Cell death by mitotic catastrophe: a molecular definition. Oncogene 23, 2825-2837.

Chauhan, S.S., Singh, A.K., Meena, S., Lohani, M., Singh, A., Arya, R.K., Cheruvu, S.H., Sarkar, J., Gayen, J.R., Datta, D., et al. (2014). Synthesis of novel beta-carboline based chalcones with high cytotoxic activity against breast cancer cells. Bioorganic & medicinal chemistry letters 24, 2820-2824.

Cory, S., and Adams, J.M. (2002). The Bcl2 family: regulators of the cellular life-or-death switch. Nature reviews Cancer 2, 647-656.

Daoud, A., Song, J., Xiao, F., and Shang, J. (2014). B-9-3, a novel beta-carboline derivative exhibits anti-cancer activity via induction of apoptosis and inhibition of cell migration in vitro. European journal of pharmacology 724, 219-230.

DeAngelis, L.M. (2001). Brain tumors. The New England journal of medicine 344, 114-123.

Deeken, J.F., and Loscher, W. (2007). The blood-brain barrier and cancer: transporters, treatment, and Trojan horses. Clinical cancer research : an official journal of the American Association for Cancer Research 13, 1663-1674.

Drucker, G., Raikoff, K., Neafsey, E.J., and Collins, M.A. (1990). Dopamine uptake inhibitory capacities of beta-carboline and 3,4-dihydro-beta-carboline analogs of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) oxidation products. Brain research 509, 125-133.

Elmore, S. (2007). Apoptosis: a review of programmed cell death. Toxicologic pathology 35, 495-516.

Esposti, M.D. (2002). The roles of Bid. Apoptosis : an international journal on programmed cell death 7, 433-440.

Esposti, M.D., Erler, J.T., Hickman, J.A., and Dive, C. (2001). Bid, a widely expressed proapoptotic protein of the Bcl-2 family, displays lipid transfer activity. Molecular and cellular biology 21, 7268-7276.

Fiszer-Maliszewska, L., and Mordarski, M. (1987). Antineoplastic activity of azacarbazoles. II. Effect of alpha-carboline and its derivatives on transplantable animal neoplasms. Archivum immunologiae et therapiae experimentalis 35, 221-224.

Friedman, H.S., Kerby, T., and Calvert, H. (2000). Temozolomide and treatment of malignant glioma. Clinical cancer research : an official journal of the American Association for Cancer Research 6, 2585-2597.

Gao, P., Tao, N., Ma, Q., Fan, W.X., Ni, C., Wang, H., and Qin, Z.H. (2014). DH332, a Synthetic beta-Carboline Alkaloid, Inhibits B Cell Lymphoma Growth by Activation of the Caspase Family. Asian Pacific journal of cancer prevention : APJCP 15, 3901-3906.

Hardy, K. (1999). Apoptosis in the human embryo. Reviews of reproduction 4, 125-134.

Helbecque, N., Moquin, C., Bernier, J.L., Morel, E., Guyot, M., and Henichart, J.P. (1987). Grossularine-1 and grossularine-2, alpha carbolines from Dendrodoa grossularia, as possible intercalative agents. Cancer biochemistry biophysics 9, 271-279.

Jackson, R.J., Fuller, G.N., Abi-Said, D., Lang, F.F., Gokaslan, Z.L., Shi, W.M., Wildrick, D.M., and Sawaya, R. (2001). Limitations of stereotactic biopsy in the initial management of gliomas. Neuro-oncology 3, 193-200.

Jaqaman, K., King, E.M., Amaro, A.C., Winter, J.R., Dorn, J.F., Elliott, H.L., McHedlishvili, N., McClelland, S.E., Porter, I.M., Posch, M., et al. (2010). Kinetochore alignment within the metaphase plate is regulated by centromere stiffness and microtubule depolymerases. The Journal of cell biology 188, 665-679.

Kalinina, J., Peng, J., Ritchie, J.C., and Van Meir, E.G. (2011). Proteomics of gliomas: initial biomarker discovery and evolution of technology. Neuro-oncology 13, 926-942.

Kastan, M.B., and Bartek, J. (2004). Cell-cycle checkpoints and cancer. Nature 432, 316-323.

Kerr, J.F., Wyllie, A.H., and Currie, A.R. (1972). Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. British journal of cancer 26, 239-257.

Kroemer, G., Galluzzi, L., and Brenner, C. (2007). Mitochondrial membrane permeabilization in cell death. Physiological reviews 87, 99-163.

Krysko, D.V., Vanden Berghe, T., D'Herde, K., and Vandenabeele, P. (2008). Apoptosis and necrosis: detection, discrimination and phagocytosis. Methods (San Diego, Calif) 44, 205-221.

Li, H., Zhu, H., Xu, C.J., and Yuan, J. (1998). Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94, 491-501.

Lin, Y., Xia, X., Yao, R., Ni, L., Hu, J., Guo, W., and Zhu, B. (2014). Synthesis and in vitro biological evaluation of hybrids from tetrahydro-beta-carboline and hydroxylcinnamic acid as antitumor carcinoma agents. Chemical & pharmaceutical bulletin 62, 343-349.

Louis, D.N., Ohgaki, H., Wiestler, O.D., Cavenee, W.K., Burger, P.C., Jouvet, A., Scheithauer, B.W., and Kleihues, P. (2007). The 2007 WHO classification of tumours of the central nervous system. Acta neuropathologica 114, 97-109.

Ly, J.D., Grubb, D.R., and Lawen, A. (2003). The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update. Apoptosis : an international journal on programmed cell death 8, 115-128.

Maddika, S., Ande, S.R., Panigrahi, S., Paranjothy, T., Weglarczyk, K., Zuse, A., Eshraghi, M., Manda, K.D., Wiechec, E., and Los, M. (2007). Cell survival, cell death and cell cycle pathways are interconnected: implications for cancer therapy. Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy 10, 13-29.

Malumbres, M., and Barbacid, M. (2001). To cycle or not to cycle: a critical decision in cancer. Nature reviews Cancer 1, 222-231.

Martin, A., Odajima, J., Hunt, S.L., Dubus, P., Ortega, S., Malumbres, M., and Barbacid, M. (2005). Cdk2 is dispensable for cell cycle inhibition and tumor suppression mediated by p27(Kip1) and p21(Cip1). Cancer cell 7, 591-598.

Muller, P.A., and Vousden, K.H. (2013). p53 mutations in cancer. Nature cell biology 15, 2-8.

Oda, T., Lee, J.S., Sato, Y., Kabe, Y., Sakamoto, S., Handa, H., Mangindaan, R.E., and Namikoshi, M. (2009). Inhibitory effect of N,N-didesmethylgrossularine-1 on inflammatory cytokine production in lipopolysaccharide-stimulated RAW 264.7 cells. Marine drugs 7, 589-599.

Ostrom, Q.T., Gittleman, H., 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 in 2006-2010. Neuro-oncology 15 Suppl 2, ii1-56.

Pardridge, W.M. (2002). Drug and gene delivery to the brain: the vascular route. Neuron 36, 555-558.

Park, W.H., Seol, J.G., Kim, E.S., Hyun, J.M., Jung, C.W., Lee, C.C., Kim, B.K., and Lee, Y.Y. (2000). Arsenic trioxide-mediated growth inhibition in MC/CAR myeloma cells via cell cycle arrest in association with induction of cyclin-dependent kinase inhibitor, p21, and apoptosis. Cancer research 60, 3065-3071.

Riedl, S.J., and Shi, Y. (2004). Molecular mechanisms of caspase regulation during apoptosis. Nature reviews Molecular cell biology 5, 897-907.

Schafer, K.A. (1998). The cell cycle: a review. Veterinary pathology 35, 461-478.

Sliwinska, M.A., Mosieniak, G., Wolanin, K., Babik, A., Piwocka, K., Magalska, A., Szczepanowska, J., Fronk, J., and Sikora, E. (2009). Induction of senescence with doxorubicin leads to increased genomic instability of HCT116 cells. Mechanisms of ageing and development 130, 24-32.

Song, H., Liu, Y., Liu, Y., Wang, L., and Wang, Q. (2014). Synthesis and antiviral and fungicidal activity evaluation of beta-carboline, dihydro-beta-carboline, tetrahydro-beta-carboline alkaloids, and their derivatives. Journal of agricultural and food chemistry 62, 1010-1018.

Strober, W. (2001). Trypan blue exclusion test of cell viability. Current protocols in immunology / edited by John E Coligan [et al] Appendix 3, Appendix 3B.

Stupp, R., Mason, W.P., van den Bent, M.J., Weller, M., Fisher, B., Taphoorn, M.J., Belanger, K., Brandes, A.A., Marosi, C., Bogdahn, U., et al. (2005). Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. The New England journal of medicine 352, 987-996.

Stupp, R., Tonn, J.C., Brada, M., and Pentheroudakis, G. (2010). High-grade malignant glioma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Annals of oncology : official journal of the European Society for Medical Oncology / ESMO 21 Suppl 5, v190-193.

Tait, S.W., and Green, D.R. (2010). Mitochondria and cell death: outer membrane permeabilization and beyond. Nature reviews Molecular cell biology 11, 621-632.

Tsai, J.Y., Hung, C.M., Bai, S.T., Huang, C.H., Chen, W.C., Chung, J.G., Kuo, S.C., Way, T.D., and Huang, L.J. (2010a). Induction of apoptosis by HAC-Y6, a novel microtubule inhibitor, through activation of the death receptor 4 signaling pathway in human hepatocellular carcinoma cells. Oncology reports 24, 1169-1178.

Tsai, J.Y., Lin, Y.C., Hsu, M.H., Kuo, S.C., and Huang, L.J. (2010b). Synthesis and cytotoxicity of 1,6,8,9-substituted alpha-carboline derivatives. The Kaohsiung journal of medical sciences 26, 593-602.

Verhoven, B., Schlegel, R.A., and Williamson, P. (1995). Mechanisms of phosphatidylserine exposure, a phagocyte recognition signal, on apoptotic T lymphocytes. The Journal of experimental medicine 182, 1597-1601.

Vermeulen, K., Van Bockstaele, D.R., and Berneman, Z.N. (2003). The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell proliferation 36, 131-149.

Vitale, I., Galluzzi, L., Castedo, M., and Kroemer, G. (2011). Mitotic catastrophe: a mechanism for avoiding genomic instability. Nature reviews Molecular cell biology 12, 385-392.

Walczak, C.E., Cai, S., and Khodjakov, A. (2010). Mechanisms of chromosome behaviour during mitosis. Nature reviews Molecular cell biology 11, 91-102.

Wang, D., and Lippard, S.J. (2005). Cellular processing of platinum anticancer drugs. Nature reviews Drug discovery 4, 307-320.

Wen, P.Y., and Kesari, S. (2008). Malignant gliomas in adults. The New England journal of medicine 359, 492-507.

Wilne, S.H., Ferris, R.C., Nathwani, A., and Kennedy, C.R. (2006). The presenting features of brain tumours: a review of 200 cases. Archives of disease in childhood 91, 502-506.

Wong, R.S. (2011). Apoptosis in cancer: from pathogenesis to treatment. Journal of experimental & clinical cancer research : CR 30, 87.
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