Responsive image
博碩士論文 etd-1101105-172551 詳細資訊
Title page for etd-1101105-172551
論文名稱
Title
熱休克與視網酸誘發惡性腦瘤細胞凋亡之探討
Induction of apoptosis in malignant brain tumor cell by heat shock and all trans retinoic acid
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
60
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2005-10-17
繳交日期
Date of Submission
2005-11-01
關鍵字
Keywords
熱休克、視網酸、惡性腦瘤、細胞凋亡
heat shock, ATRA, malignant brain tumor, apoptosis
統計
Statistics
本論文已被瀏覽 5675 次,被下載 14
The thesis/dissertation has been browsed 5675 times, has been downloaded 14 times.
中文摘要
癌症為台灣十大死因之首,雖然腦瘤在惡性腫瘤佔少數,但治癒率不佳。多形性神經膠質母細胞瘤(Glioblastoma multiforme,GBM)是成人腦部最常見惡性的腫瘤。前人利用熱休克或對式視網酸(all trans retinoic acid,ATRA)分別在其他癌細胞株證實可以引發細胞凋亡及細胞週期停滯。在惡性膠質細胞瘤並無相關研究。因此本實驗利用人類腦瘤細胞株GBM8401為研究材料,先以43℃ 處理30 min後再加入ATRA。以生長曲線分析經處理後的GBM8401,發現其生長抑制達50%。而以代謝活性分析(MTT assay)分析約降低50%。進一步以反轉錄聚合酵素連鎖反應及西方轉漬法分析細胞凋亡相關基因及蛋白質,發現GBM8401細胞置於43℃ 30分鐘後加入ATRA作用15分鐘至8小時之間,p21、p27、pro-caspase 3、phospho-JNK、phospho-p38即有增加表現的情形。以免疫細胞化學染色分析在處理8小時後,可發現在細胞核內phospho-p53過量表現的情形。實驗結果表示熱休克後ATRA處理可促進細胞凋亡的過程,達到抑制GBM8401生長的效果。
Abstract
Cancer has become the first among the 10 major death causing factors in Taiwan. Glioblastoma multiforme (GBM) is the most common malignant tumor in adult human brain tumors. Previously, heat shock or all trans retinoic acid (ATRA) treatment has been shown to be effective in inducing cell apoptosis and cell cycle arrest in several cancer cell lines. In this study, human brain tumor cell line GBM8401 was exposed to 43℃ for 30 min followed by incubation with ATRA. The treatment resulted in up to 50% inhibition of cell growth rate and 50% reduction of cell survival rate . Analysis of cell apoptosis related gene expression and protein expression with RT-PCR and Western blot has showen that p21, p27, pro-caspase 3, phospho-JNK and phospho-p38 were overexpressed after treatment of tumor cells with 43℃ for 30 min followed by addition of ATRA for 15 min to 8 hr. The immunocytochemistry assay revealed that overexpression of phospho-p53 in the nuclei after tumor cells were treated with 43℃ for 30 min followed by addition of ATRA for 8 hr. Results from this study show that treating tumor cells with heat shock before incubation with ATRA will enhance cell apoptosis and inhibit cell growth.
目次 Table of Contents
中文摘要…………………………………………………1
英文摘要…………………………………………………2
壹、前言…………………………………………………3
貳、研究目的……………………………………………15
參、材料與方法…………………………………………16
肆、實驗結果與討論……………………………………24
伍、結論…………………………………………………33
陸、參考文獻……………………………………………34
柒、圖表…………………………………………………40
參考文獻 References
林瑞雄。1995。原發性腦瘤流行病學研究 政府研究資訊系統。

腦瘤之診斷與治療共識。2004。國家衛生研究院。

Abu J, Batuwangala M, Herbert K, Symonds P.(2005)Retinoic acid and retinoid receptors: potential chemopreventive and therapeutic role in cervical cancer. Lancet Oncol. 6, 712-720.

Alsayed Y, Uddin S, Mahmud N, Lekmine F, Kalvakolanu DV, Minucci S, Bokoch G, Platanias LC.(2001)Activation of Rac1 and the p38 mitogen-activated protein kinase pathway in response to all-trans-retinoic acid. J Biol Chem 276, 4012-4019.

Andela V B, Rosier R N.(2004)The proteosome inhibitor MG132 attenuates Retinoic Acid Receptor trans-activation and enhances trans-repression of Nuclear Factor κB. Potential relevance to chemo-preventive interventions with retinoids. Mol Cancer 22, 3-8.

Arce F, Gatjens-Boniche O, Vargas E, Valverde B, Diaz C.(2005)Apoptotic events induced by naturally occurring retinoids ATRA and 13-cis retinoic acid on human hepatoma cell lines Hep3B and HepG2. Cancer Lett 229, 271-281.

Azrak RG, Cao S, Slocum HK, Toth K, Durrani FA, Yin MB, Pendyala L, Zhang W, McLeod HL, Rustum YM.(2004)Therapeutic synergy between irinotecan and 5-fluorouracil against human tumor xenografts.Clin Cancer Res. 10, 1121-1129.

Bardon S, Razanamahefa L.(1998)Retinoic acid suppresses insulin-induced cell growth and cyclin D1 gene expression in human breast cancer cells. Int J Oncol. 12, 355-359.

Beere HM.(2001) Stressed to death: regulation of apoptotic signaling pathways by the heat shock proteins. Sci STKE. 31 RE1.

Bhaskara VK, Panigrahi M, Challa S, Babu PP.(2005)Comparative status of activated ERK1/2 and PARP cleavage in human gliomas. Neuropathology 25, 48-53.

Blomberg I, Hoffmann I.(1999)Ectopic expression of Cdc25A accelerates the G(1)/S transition and leads to premature activation of cyclin E- and cyclin A-dependent kinases. Mol Cell Biol.19, 6183-6194.

Callari D, Sinatra F, Paravizzini G, Libra M, Emmanuele G, Fiore B, Pezzino MF, Rasa D, Mazzarino MC, D'Alessandro N, Travali S.(2003)All trans retinoic acid sensitizes colon cancer cells to hyperthermia cytotoxic effects. Int J Oncol. 23, 181-188.

Carson DA, Ribeiro JM.(1993)Apoptosis and disease. Lancet. 341, 1251-1254.

Chen CF, Goyette P , Lohnes D.(2004)RARγ acts as a tumor suppressor in mouse keratinocytes. Oncogene 8, 5350-5359.

Cobb MH.(1999)MAP kinase pathways. Prog Biophys Mol Biol.71:479-500.

Chu PW, Cheung WM, Kwong YL.(2003) Differential effects of 9-cis, 13-cis and all-trans retinoic acids on the neuronal differentiation of human neuroblastoma cells. Neuroreport 14, 1935-1939.

Cory S, Huang DC, Adams JM.(2003)The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene. 22:8590-607.

Donovan, M , Cotter, T.G.(2004)Control of mitochondrial integrity by Bcl-2 family members and caspase-indendent cell death. Biochim Biophys.Acta.1644:133-147.

Dreyfus DH, Nagasawa M, Gelfand EW , Ghoda LY.(2005)Modulation of p53 activity by IκBα: Evidence suggesting a common phylogeny between NF-κB and p53 transcription factors. BMC Immunol 21, 6-12.

Franklin TB, Krueger-Naug AM, Clarke DB, Arrigo AP, Currie RW.(2005) The role of heat shock proteins Hsp70 and Hsp27 in cellular protection of the central nervous system. Int J Hyperthermia. 21, 379-92.

Gerschenson LE, Rotello RJ.(1992) Apoptosis: a different type of cell death. FASEB J 6, 2450-2455.

Gerl R, Vaux DL.(2005)Apoptosis in the development and treatment of cancer. Carcinogenesis 26, 263-270.

Golias CH, Charalabopoulos A, Charalabopoulos K.(2004) Cell proliferation and cell cycle control .Int J Clin Pract.58, 1134-1141.

Grimison B, Langan TA, Sclafani RA.(2000)p16Ink4a tumor suppressor function in lung cancer cells involves cyclin-dependent kinase 2 inhibition by Cip/Kip protein redistribution. Cell Growth Differ. 11, 507-515.

Gumireddy K, Sutton LN, Phillips PC, Reddy CD.(2003) All-trans-retinoic acid-induced apoptosis in human medulloblastoma: activation of caspase-3/poly(ADP-ribose) polymerase 1 pathway. Clin Cancer Res. 9, 4052-4059.

Hayashi T, Abe K. (2004) Ischemic neuronal cell death and organellae damage. Neurol Res. 26, 827-834.
Helmbrecht K, Zeise E, and Rensing L. (2000) Chaperone in cell cycle regulation and mitogenic signal transduction. Cell Prolif. 33: 341–365.

Houston A, O'Connell J.(2004) The Fas signalling pathway and its role in the pathogenesis of cancer. Curr Opin Pharmacol. 4:321-326.

Jin Z, El-Deiry WS.(2005)Overview of cell death signaling pathways. Cancer Biol Ther. 4,139-63.

Kaba SE, Kyritsis AP, Conrad C, Gleason MJ, Newman R, Levin VA, Yung WK.(1997) The treatment of recurrent cerebral gliomas with all-trans-retinoic acid(tretinoin). J Neurooncol.34, 145-151.

Kania J, Baranska A, Guzdek A.(2003) Cytokine action and oxidative stress response in differentiated neuroblastoma SH-SY5Y cells.Acta Biochim Pol.50, 659-66.

Komata T, Kanzawa T, Nashimoto T, Aoki H, Endo S, Nameta M, Takahashi H, Yamamoto T, Kondo S, Tanaka R.(2004) Mild heat shock induces autophagic growth arrest, but not apoptosis in U251-MG and U87-MG human malignant glioma cells. J Neurooncol. 68, 101-111.

Kuhl NM, Kunz J, Rensing L.(2000)Heat shock-induced arrests in different cell cycle phases of rat C6-glioma cells are attenuated in heat shock-primed thermotolerant cells. Cell Prolif. 33, 147-166.

Kuo PC, Liu HF, and Chao JI.(2004) Survivin and p53 modulate quercetin-induced cell growth inhibition and apoptosis in Human lung carcinoma cells. J Biol Chem. 279, 55875-55885.

Langlois A, Lee S, Kim DS, Dirks PB, Rutka JT.(2002)p16(ink4a) and retinoic acid modulate rhoA and GFAP expression during induction of a stellate phenotype in U343 MG-A astrocytoma cells. Glia. 40, 85-94.

Lania L, Majello B, Napolitano G.(1999) Transcriptional control by cell-cycle regulators. J Cell Physiol. 179,134-141.

Lu TS, Chen HW, Huang MH, Wang SJ, Yang RC.(2004) Heat shock treatment protects osmotic stress-induced dysfunction of the blood-brain barrier through preservation of tight junction proteins. Cell Stress Chaperones 9, 369-377.

Lynne S. Cox.(1997) Multiple pathways control cell growth and transformation: overlapping and independent activities of p53 and p21Cip1/WAF1/Sdi1 J Pathol. 183:134-140.

Miller WH .(1998) The emerging role of retinoids and retinoic acid metabolism blocking agents in the treatment of cancer.Cancer 83, 1471-82.

Nunez G, Benedict MA, Hu Y, Inohara N.(1998) Caspases: the proteases of the apoptotic pathway. Oncogene 17, 3237-3245.

Oliferenko S, Balasubramanian MK.(2001)Cell cycle: the Flp side of Cdc14. Curr Biol. 11, 872-874.

Ortiz MA, Lopez-Hernandez FJ, Bayon Y, Pfahl M, Piedrafita FJ.(2001) Retinoid-related molecules induce cytochrome c release and apoptosis through activation of c-Jun NH(2)-terminal kinase/p38 mitogen-activated protein kinases. Cancer Res. 61, 8504-8512.

Pardridge WM.(2003)Blood-brain barrier drug targeting: the future of brain drug development. Mol Interv. 3:90-105.

Park KH, Lee J, Yoo CG, Kim YW, Han SK, Shim YS, Kim SK, Wang KC, Cho BK, Lee CT.(2004) Application of p27 gene therapy for human malignant glioma potentiated by using mutant p27. J Neurosurg. 101, 505-510.

Parney IF, Chang SM.(2003) Current chemotherapy for glioblastoma. Cancer J. 9, 149-56.

Pirozzoli MC, Marino C, Lovisolo GA, Laconi C, Mosiello L, Negroni A.(2003) Effects of 50 Hz electromagnetic field exposure on apoptosis and differentiation in a neuroblastoma cell line. Bioelectromagnetics 24, 510-6.

Wang Q, Wieder R.(2004) All-trans retinoic acid potentiates Taxotere-induced cell death mediated by Jun N-terminal kinase in breast cancer cells. Oncogen 23, 426-433

Ran LW, Tan SS, Wang WJ, Qiu S, Lei XB, Zeng WH.(2005) Effects of all-trans-retinoic acid, acitretin and tazarotene on apoptosis and Bax/Bcl-2 expressions of human melanoma cells A375 and the significance. Di Yi Jun Yi Da Xue Xue Bao 25, 972-978.

Reynolds CP, Lemons RS.(2001)Retinoid therapy of childhood cancer. Hematol Oncol Clin North Am.15, 867-910.

Ritossa F.(1962)A new pattern induced by temperature shock and DNP in Drosophila .Experientia 13, 571-573.

Searle J, Kerr JF, Bishop CJ.(1982) Necrosis and apoptosis: distinct modes of cell death with fundamentally different significance. Pathol Annu.17, 229-59.

Shah MA, Schwartz GK.(2001)Cell cycle-mediated drug resistance: an emerging concept in cancer therapy. Clin Cancer Res. 7, 2168-2181.

Sharpe JC, Arnoult, D, Youle, RJ.(2004) Control of mitochondrial permeability by Bcl-2 family members. Biochimica et Biophysica Acta (BBA)/Molecular Cell Research.1644, 107-113.

Simoni D, Tolomeo M.(2001)Retinoids, apoptosis and cancer. Curr Pharm Des. 7:1823-1837.

Slee EA, Adrain C, Martin SJ.(2001) Executioner caspase-3, -6, and -7 perform distinct, non-redundant roles during the demolition phase of apoptosis. J Biol Chem. 276, 7320-7326.

Steinbach JP, Weller M.(2004) Apoptosis in Gliomas: Molecular mechanisms and therapeutic implications. J Neurooncol. 70, 247-256.

Tallman MS.(1996) Differentiating therapy in acute myeloid leukemia. Leukemia 10, 1262-1268.

Tannoch VJ, Hinds PW, Tsai LH.(2000) Cell cycle control. Adv Exp Med Biol.465, 127-140.

Tissieres A, Mitchell HK, and Tracy VM. (1974) Protein synthesis in salivary gland of Drosophila melanogaster. Relation to chromosome puffs. J. Mol Biol. 84, 389-398.

Tomida A, Suzuki H, Kim HD, Tsuruo T.(1996)Glucose-regulated stresses cause decreased expression of cyclin D1 and hypophosphorylation of retinoblastoma protein in human cancer cells. Oncogene 13, 2699-2705.

Turco MC, Romano MF, Petrella A, Bisogni R, Tassone P, Venuta S.(2004) NF-kappaB/Rel-mediated regulation of apoptosis in hematologic malignancies and normal hematopoietic progenitors. Leukemia 18, 11-17.

Uesugi S, Yamashita K, Nakashima K, Ito H.(1998) Apoptotic cell death induced by local brain hyperthermia in a rat glioma model. Acta Neuropathol. 96, 351-356.

Wajant H.(2004)TRAIL and NFkappaB signaling--a complex relationship. Vitam Horm 67, 101-32.

Wang Q, Wieder R.(2004)All-trans retinoic acid potentiates Taxotere-induced cell death mediated by Jun N-terminal kinase in breast cancer cells. Oncogene 23:426-433.

Wyllie AH, Beattie GJ, Hargreaves AD.(1981) Chromatin changes in apoptosis. Histochem J. 13, 681-692.

Yenari MA, Liu J, Zheng Z, Vexler ZS, Lee JE, Giffard RG.(2005) Antiapoptotic and Anti-inflammatory Mechanisms of Heat-Shock Protein Protection. Ann N Y Acad Sci.1053, 74-83.

Yuan J, Yankner BA.(2000) Apoptosis in the nervous system. Nature 407, 802-809.

Zang C, Wachter M, Liu H, Posch MG, Fenner MH, Stadelmann C, von Deimling A, Possinger K, Black KL, Koeffler HP, Elstner E.(2003) Ligands for PPARgamma and RAR cause induction of growth inhibition and apoptosis in human glioblastomas. J Neurooncol. 65, 107-118.

Zhang H, Rosdahl I.(2004) Expression profiles of p53, p21, bax and bcl-2 proteins in all-trans-retinoic acid treated primary and metastatic melanoma cells. Int J Oncol. 25, 303-8.

Zhang WL, Tsuneishi S, Nakamura H.(2001) Induction of heat shock proteins and its effects on glial differentiation in rat C6 glioblastoma cells.Kobe J Med Sci. 47, 77-95.

Zylicz M , King F W and Wawrzynow A.(2001) Hsp70 interactions with the p53 tumour suppressor protein. EMBO 20, 4634–4638.
電子全文 Fulltext
本電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。
論文使用權限 Thesis access permission:校內一年後公開,校外永不公開 campus withheld
開放時間 Available:
校內 Campus: 已公開 available
校外 Off-campus:永不公開 not available

您的 IP(校外) 位址是 44.222.63.67
論文開放下載的時間是 校外不公開

Your IP address is 44.222.63.67
This thesis will be available to you on Indicate off-campus access is not available.

紙本論文 Printed copies
紙本論文的公開資訊在102學年度以後相對較為完整。如果需要查詢101學年度以前的紙本論文公開資訊,請聯繫圖資處紙本論文服務櫃台。如有不便之處敬請見諒。
開放時間 available 已公開 available

QR Code