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系統識別號 U0026-0102201014340300
論文名稱(中文) 合併處理放射線與三氧化二砷對骨肉瘤的細胞毒性機制探討
論文名稱(英文) Study on the cytotoxic mechanism of irradiation combined with arsenic trioxide in osteosarcoma
校院名稱 成功大學
系所名稱(中) 環境醫學研究所
系所名稱(英) Institute of Environmental and Occupational Health
學年度 98
學期 1
出版年 99
研究生(中文) 林葦
研究生(英文) Wei Lin
學號 S7696107
學位類別 碩士
語文別 中文
論文頁數 89頁
口試委員 口試委員-何元順
口試委員-潘敏雄
口試委員-何聖佑
指導教授-王應然
中文關鍵字 骨肉瘤  放射線  三氧化二砷  G2/M期停滯  協同效應  細胞凋亡  細胞自體吞噬 
英文關鍵字 Osteosarcoma  Ionizing radiation  arsenic trioxide  Synergistic effect  G2/M arrest  apoptosis  autophagy 
學科別分類
中文摘要 骨肉瘤(osteosarcoma)是一種最常見的惡性骨腫瘤,大約佔所有惡性骨腫瘤的45%。將患肢的部位做截肢是骨肉瘤的主要治療方式,但是手術後的存活率還是相當低。在臨床上大約50~60%的癌症病人都選擇放射線治療。但是骨肉瘤細胞會對放射線產生抗性,因此要尋找一種能有效增加放射線療效的藥劑便顯得重要且迫切。臨床上已經證實對於治療急性骨髓白血病,三氧化二砷是一個很有效的抗癌藥劑。本篇研究最主要的目的在探討當合併放射線與三氧化二砷治療時,是否會增加對骨肉瘤細胞的毒性及其毒性機制探討。研究設計選取人類骨肉瘤細胞株-HOS,分別給予暴露不同劑量的三氧化二砷以及放射線。實驗中,首先評估細胞經處理後的細胞存活率,利用Trypan blue 和clonogenic assay來做測試;利用劑量效應分析軟體分析協同效應;使用流式細胞儀分析細胞週期、有絲分裂指數(mitotic index)、活性氧化物含量、粒線體膜電位變化,及藉由annexin V-FITC 來分析早期細胞凋亡的現象;Comet assay、DAPI以及Giemsa染色用來分析DNA受損程度、細胞凋亡的微核現象以及觀察mitotic cells的比例;利用acridine orange染色觀察細胞自體吞噬的酸性小泡(AVO)產生。使用穿透式電子顯微鏡觀察HOS細胞的超顯微結構。以西方墨點法探討細胞週期、細胞凋亡及自體吞噬相關蛋白的表現。由實驗結果發現,相較於單獨處理放射線或是三氧化二砷的組別,合併處理的組別效果更好。三氧化二砷可以增加HOS細胞的放射線敏感性以及與放射線合併使用會有協同效應。合併處理會造成更高的細胞毒性以及造成細胞週期G2/M期的停滯,特別是使細胞停滯在M期。此外,相較於單獨處理放射線或是三氧化二砷的組別,合併處理後會降低細胞的粒線體膜電位、延緩細胞DNA的修復、增加活性氧化物的釋放以及早期細胞凋亡和自體吞噬的百分比。在蛋白質表現方面,合併處理的組別其p21、cyclin B1、LC3 II、p62、cleaved-PARP、cleaved-caspase 3、cytochrome c及p-GSK3β表現量增加,而p-ERK, p-Akt and p-PDK1表現量下降。p-cdc2 and p-cdc25c在單獨處理放射線的組別中表現量增加,但是合併處理後表現量下降。我們也發現三氧化二砷會增加Bcl-2 and Bcl-xL蛋白的磷酸化現象。此外,前處理Z-VAD-fmk(caspase的抑制劑)以及3MA(自體吞噬的抑制劑)都會降低合併處理所引起的細胞凋亡及自體吞噬的比例。此研究證明,三氧化二砷能使對放射線有抗性的HOS細胞株更具有細胞毒性(cytotoxic)效果。且合併處理之後所誘發的死亡方式包含了細胞凋亡及自體吞噬。
英文摘要 Osteosarcma is the most common malignant neoplasm of bone. The standard treatment for osteosarcoma was amputation of the affected limb. However, the chance of recovering has been pretty low. In clinical treatment, about 50 to 60 percent of cancer patients are treated with radiation for their cancers but it is not equally effective in all cancer cells. Some cancer cells such as osteosarcoma are resistant to radiotherapy, thus prompted a search for agents which can enhance IR efficacy is urgent and important. Clinical data demonstrated that arsenic trioxide (ATO) is an effective anticancer agent for the treatment of acute promyelocytic leukemia (APL). The aim of this study is to investigate the cytotoxic mechanism of irradiation combined with arsenic trioxide (ATO) in the treatment of osteosarcoma. HOS cell lines (human osteosarcoma cells) were used in this study and they were treated with different doses of ATO and/or IR (Ionizing radiation), respectively. First, cell viability was detected by trypan blue and clonogenic assay, and the combination effect was analyzed by using CalcuSyn software;Cell cycle distribution, mitotic index, ROS, membrane potential and early apoptosis with annexin V-FITC apoptosis detection kit were detected by flow cytometry. DNA damage was detected by comet assay. Microphotograph of micronuclei formation in HOS cells by DAPI-stained were performed. We examined the mitotic arrest of HOS cells after IR and/or ATO treatment using the Giemsa staining procedure. In order to observe the expression of acidic vesicular organelle (AVO) which is an important characteristic of autophagy, cells were stained with acridine orange. Ultrastructure of HOS cells was anayzed by electron microscopy. Western blotting was performed to analyze the expression of protein related to cell cycle, apoptosis and autophagy. The results indicated that the effect of combined treatment is more significant then irradiation or arsenic alone. Arsenic trioxide enhanced radiosensitivity in HOS cells and showed synergistic effect with irradiation. The combined treatment showed a higher cytotoxic effects and G2 phase and mitotic arrest than treated alone. Furthermore, compared with the alone treatment of IR and ATO, the combined treatment could reduce mitochondrial membrane potential (ΔΨm), delayed the DNA repair and enhance the percentage of intracellular ROS, apoptosis and autophagy. The expression level of p21, cyclin B1, LC3 II, p62, cleaved-PARP, cleaved-caspase 3, cytochrome c, p-GSK3β were increased. p-ERK, p-Akt and p-PDK1 expression were decreased. p-cdc2 and p-cdc25c were increased in the alone treatment of IR but they were decreased in the combined treatment. We also observed that arsenic trioxide induced phosphorylation of Bcl-2 and Bcl-xL. In addition, pretreatment of Z-VAD-fmk (a caspase inhibitor) and 3MA (an autophagy inhibitor) decreased the combination-induced apoptosis and autophagy. Taken together, this study demonstrated that ATO increase the therapeutic efficacy of radiotherapy against HOS cells. The cell death mechanisms induced by the combined treatment include apoptosis and autophagy.
論文目次 第一章、序論............................................. 1
第二章、文獻回顧......................................... 2
第一節、惡性骨肉瘤(osteosarcoma)以及其治療方式.......... 2
第二節、放射線治療.................................... 3
第三節、放射線敏感性(Radiosensitivity)與阻抗性(Radioresistant)....................................... 4
第四節、三氧化二砷(As2O3).............................. 5
第五節、放射治療合併化學治療........................... 8
第六節、細胞週期(cell cycle).......................... 8
第七節、細胞凋亡(Apoptosis)與自體吞噬(Autophagy)........ 12
第三章、研究目的......................................... 16
第四章、研究架構......................................... 17
第五章、研究材料與方法.................................... 18
第一節 研究材料....................................... 18
(一) 細胞株........................................ 18
(二) 儀器.......................................... 18
(三) 試劑.......................................... 19
(四) 溶液.......................................... 20
第二節 研究方法....................................... 22
(一) 人類骨肉瘤細胞(HOS)培養液(medium)製備........... 22
(二) 細胞解凍...................................... 22
(三) HOS 細胞培養.................................. 22
(四) 細胞冷凍...................................... 23
(五) Trypan blue dye exclusion assay : 細胞計數.... 24
(六) clonogenic assay............................. 24
(七) 細胞週期(cell cycle)分析....................... 26
(八) 活性氧物種(ROS)分析............................ 27
(九) 粒線體膜電位 membrane potential(△Ψm)分析....... 27
(十) 彗星試驗(Comet assay)......................... 28
(十一) 螢光顯微鏡分析細胞微核現象(micronuclei formation)............................................. 30
(十二) 細胞凋亡(apoptosis)分析...................... 31
(十三) 細胞自體吞噬(Autophagy)分析................... 31
(十四) 有絲分裂指數(Mitotic index)分析............... 32
(十五) 螢光顯微鏡分析自體吞噬(Autophagy)現象.......... 33
(十六) Giemsa stain................................ 33
(十七) 電子顯微鏡................................... 34
(十八) Western blot................................ 35
(十九) 協同效應分析(Synergistic interaction)........ 40
(二十) 統計分析..................................... 40
第六章、實驗結果......................................... 41
第一節 放射線和三氧化二砷對HOS細胞的劑量及時間效應........ 41
第二節 合併放射線與三氧化二砷對HOS細胞的放射線敏感性之影響. 42
第三節 放射線合併三氧化二砷對HOS細胞的細胞週期變化........ 43
第四節 探討放射線與三氧化二砷對HOS細胞凋亡現象............ 44
第五節 分析HOS細胞其Autophagy的表現.................... 46
第六節 放射線合併三氧化二砷對HOS細胞的協同作用分析........ 49
第七節 細胞訊息傳遞路徑之相關蛋白表現.................... 49
第八節 抑制HOS cell細胞凋亡(Apoptosis)或細胞自體吞噬(Autophagy)的影響....................................... 50
第七章、討論............................................. 51
第八章、結論與建議 ........................................58
第九章、參考文獻......................................... 75
參考文獻 Adams JM, Cory S. 1998. The Bcl-2 protein family: arbiters of cell survival. Science 281: 1322-1326.
Afford S, Randhawa S. 2000. Apoptosis. Mol Pathol 53: 55-63.
Anderson. P, Aguilera. D, Pearson. M, Woo. S. 2008. Outpatient chemotherapy plus radiotherapy in sarcomas: improving cancer control with radiosensitizing agents. Cancer Control 15: 38-46.
Antman KH. 2001. Introduction: the history of arsenic trioxide in cancer therapy. Oncologist 6 Suppl 2: 1-2.
Ashby J, Tinwell H, Lefevre PA, Browne MA. 1995. The single cell gel electrophoresis assay for induced DNA damage (comet assay): measurement of tail length and moment. Mutagenesis 10: 85-90.
Baehrecke EH. 2003. Autophagic programmed cell death in Drosophila. Cell Death Differ 10: 940-945.
Barkanov AI. 1978. [Radiation therapy of malignant bone tumors]. Med Radiol (Mosk) 23: 69-77.
Basu A, Haldar S. 2003. Identification of a novel Bcl-xL phosphorylation site regulating the sensitivity of taxol- or 2-methoxyestradiol-induced apoptosis. FEBS Lett 538: 41-47.
Bauman GS, Wara WM, Ciricillo SF, Davis RL, Zoger S, Edwards MS. 1997. Primary intracerebral osteosarcoma: a case report. J Neurooncol 32: 209-213.
Bellamy COC, Malcolmson RDG, Harrison DJ, Wyllie AH. 1995. Cell death in health and disease: the biology and regulation of apoptosis. Seminars in Cancer Biology 6: 3-16.
Bielack SS, Carrle D, Hardes J, Schuck A, Paulussen M, Bielack SS, et al. 2008. Bone tumors in adolescents and young adults. Curr Treat Options Oncol 9: 67-80.
Biermann JS, Adkins D. 2007. Bone cancer. Journal of the National Comprehensive Cancer Network 5: 420-437.
Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, Overvatn A, et al. 2005. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 171: 603-614.
Bommareddy A, Hahm ER, Xiao D, Powolny AA, Fisher AL, Jiang Y, et al. 2009. Atg5 regulates phenethyl isothiocyanate-induced autophagic and apoptotic cell death in human prostate cancer cells. Cancer Res 69: 3704-3712.
Broyn. 1975. Kinetics of cell proliferation and cell loss in the peripheral and central parts of Walker tumours growing in rats and nude mice. Virchows Arch B Cell Pathol 18: 181-191.
Bucher N, Britten CD. 2008. G2 checkpoint abrogation and checkpoint kinase-1 targeting in the treatment of cancer. Br J Cancer 98: 523-528.
Bulavin DV, Amundson SA, Fornace AJ. 2002. p38 and Chk1 kinases: different conductors for the G(2)/M checkpoint symphony. Curr Opin Genet Dev 12: 92-97.
Cavigelli M, Li WW, Lin A, Su B, Yoshioka K, Karin M. 1996. The tumor promoter arsenite stimulates AP-1 activity by inhibiting a JNK phosphatase. Embo J 15: 6269-6279.
Chalmers AJ, Ruff EM, Martindale C, Lovegrove N, Short SC. 2009. Cytotoxic effects of temozolomide and radiation are additive- and schedule-dependent. Int J Radiat Oncol Biol Phys 75: 1511-1519.
Cheng JQ, Lindsley CW, Cheng GZ, Yang H, Nicosia SV. 2005. The Akt/PKB pathway: molecular target for cancer drug discovery. Oncogene 24: 7482-7492.
Cheng Y, Qiu F, Ikejima T. 2009. Molecular mechanisms of oridonin-induced apoptosis and autophagy in murine fibrosarcoma L929 cells. Autophagy 5: 430-431.
Chiarugi V, Magnelli L, Cinelli M. 1998. Role of p53 mutations in the radiosensitivity status of tumor cells. Tumori 84: 517-520.
Chiu HW, Ho SY, Guo HR, Wang YJ. 2009. Combination treatment with arsenic trioxide and irradiation enhances autophagic effects in U118-MG cells through increased mitotic arrest and regulation of PI3K/Akt and ERK1/2 signaling pathways. 472-483.
Chun YJ, Park IC, Park MJ, Woo SH, Hong SI, Chung HY, et al. 2002. Enhancement of radiation response in human cervical cancer cells in vitro and in vivo by arsenic trioxide (As2O3). FEBS Lett 519: 195-200.
Colosetti P, Puissant A, Robert G, Luciano F, Jacquel A, Gounon P, et al. 2009. Autophagy is an important event for megakaryocytic differentiation of the chronic myelogenous leukemia K562 cell line. Autophagy 5: 1092-1098.
Coustan-Smith E, Kitanaka A, Pui CH, McNinch L, Evans WE, Raimondi SC, et al. 1996. Clinical relevance of BCL-2 overexpression in childhood acute lymphoblastic leukemia. Blood 87: 1140-1146.
Davison K, Mann KK, Miller WH, Jr. 2002. Arsenic trioxide: mechanisms of action. Semin Hematol 39: 3-7.
de Bruin EC, Medema JP. 2008. Apoptosis and non-apoptotic deaths in cancer development and treatment response. Cancer Treat Rev 34: 737-749.
Douer D, Tallman MS. 2005. Arsenic trioxide: new clinical experience with an old medication in hematologic malignancies. J Clin Oncol 23: 2396-2410.
Du L, Lyle CS, Chambers TC. 2005. Characterization of vinblastine-induced Bcl-xL and Bcl-2 phosphorylation: evidence for a novel protein kinase and a coordinated phosphorylation/dephosphorylation cycle associated with apoptosis induction. Oncogene 24: 107-117.
Dudek H, Datta SR, Franke TF, Birnbaum MJ, Yao R, Cooper GM, et al. 1997. Regulation of neuronal survival by the serine-threonine protein kinase Akt. Science 275: 661-665.
Edinger AL, Thompson CB. 2004. Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol 16: 663-669.
Eisenberg-Lerner A, Kimchi A. 2009. The paradox of autophagy and its implication in cancer etiology and therapy. Apoptosis 14: 376-391.
Elledge SJ. 1996. Cell cycle checkpoints: preventing an identity crisis. Science 274: 1664-1672.
Eymin B, Leduc C, Coll JL, Brambilla E, Gazzeri S. 2003. p14ARF induces G2 arrest and apoptosis independently of p53 leading to regression of tumours established in nude mice. Oncogene 22: 1822-1835.
Fairbairn DW, Olive PL, O'Neill KL. 1995. The comet assay: a comprehensive review. Mutat Res 339: 37-59.
Fukuda K, Sakakura C, Miyagawa K, Kuriu Y, Kin S, Nakase Y, et al. 2004. Differential gene expression profiles of radioresistant oesophageal cancer cell lines established by continuous fractionated irradiation. Br J Cancer 91: 1543-1550.
Gaitan-Yanguas M. 1981. A study of the response of osteogenic sarcoma and adjacent normal tissues to radiation. Int J Radiat Oncol Biol Phys 7: 593-595.
Gewirtz DA, Hilliker ML, Wilson EN. 2009. Promotion of autophagy as a mechanism for radiation sensitization of breast tumor cells. Radiother Oncol 92: 323-328.
Girdhani S, Bhosle SM, Thulsidas SA, Kumar A, Mishra KP. 2005. Potential of radiosensitizing agents in cancer chemo-radiotherapy. J Cancer Res Ther 1: 129-131.
Gonzalez-Polo RA, Boya P, Pauleau AL, Jalil A, Larochette N, Souquere S, et al. 2005. The apoptosis/autophagy paradox: autophagic vacuolization before apoptotic death. J Cell Sci 118: 3091-3102.
Haimovitz-Friedman A. 1998. Radiation-induced signal transduction and stress response. Radiat Res 150: S102-108.
Hartwell LH, Weinert TA. 1989. Checkpoints: controls that ensure the order of cell cycle events. Science 246: 629-634.
Haykal J, Fernainy P, Itani W, Haddadin M, Geara F, Smith C, et al. 2008. Radiosensitization of EMT6 mammary carcinoma cells by 2-benzoyl-3-phenyl-6,7-dichloroquinoxaline 1,4-dioxide. Radiother Oncol 86: 412-418.
Hengartner MO. 2000. The biochemistry of apoptosis. Nature 407: 770-776.
Ho SY, Chen WC, Chiu HW, Lai CS, Guo HR, Wang YJ, et al. 2009. Combination treatment with arsenic trioxide and irradiation enhances apoptotic effects in U937 cells through increased mitotic arrest and ROS generation. Chem Biol Interact 179: 304-313.
Ho SY, Huang PC, Guo HR, Chang WH, Chen RJ, Wei BL, et al. 2006. Mechanisms of apoptosis induction and cell cycle regulation in irradiated leukemia U937 cells and enhancement by arsenic trioxide. Radiat Res 165: 390-399.
Hu H, Chai Y, Wang L, Zhang J, Lee HJ, Kim SH, et al. 2009. Pentagalloylglucose induces autophagy and caspase-independent programmed deaths in human PC-3 and mouse TRAMP-C2 prostate cancer cells. Mol Cancer Ther 8: 2833-2843.
Hung JY, Hsu YL, Li CT, Ko YC, Ni WC, Huang MS, et al. 2009. 6-Shogaol, an Active Constituent of Dietary Ginger, Induces Autophagy by Inhibiting the AKT/mTOR Pathway in Human Non-Small Cell Lung Cancer A549 Cells. J Agric Food Chem.
Hwang A, Muschel RJ. 1998. Radiation and the G2 phase of the cell cycle. Radiat Res 150: S52-59.
Iwama K, Nakajo S, Aiuchi T, Nakaya K. 2001. Apoptosis induced by arsenic trioxide in leukemia U937 cells is dependent on activation of p38, inactivation of ERK and the Ca2+-dependent production of superoxide. Int J Cancer 92: 518-526.
Jackson SP. 2002. Sensing and repairing DNA double-strand breaks. Carcinogenesis 23: 687-696.
Jenkin RD, Allt WE, Fitzpatrick PJ. 1972. Osteosarcoma. An assessment of management with particular reference to primary irradiation and selective delayed amputation. Cancer 30: 393-400.
Jiang JD, Davis AS, Middleton K, Ling YH, Perez-Soler R, Holland JF, et al. 1998. 3-(Iodoacetamido)-benzoylurea: a novel cancericidal tubulin ligand that inhibits microtubule polymerization, phosphorylates bcl-2, and induces apoptosis in tumor cells. Cancer Res 58: 5389-5395.
Jin HO, Seo SK, Woo SH, Lee HC, Kim ES, Yoo DH, et al. 2008. A combination of sulindac and arsenic trioxide synergistically induces apoptosis in human lung cancer H1299 cells via c-Jun NH2-terminal kinase-dependent Bcl-xL phosphorylation. Lung Cancer 61: 317-327.
Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, et al. 2000. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J 19: 5720-5728.
Kang YH, Lee SJ. 2008. Role of p38 MAPK and JNK in enhanced cervical cancer cell killing by the combination of arsenic trioxide and ionizing radiation. Oncol Rep 20: 637-643.
Kanzawa T, Zhang L, Xiao L, Germano IM, Kondo Y, Kondo S. 2005. Arsenic trioxide induces autophagic cell death in malignant glioma cells by upregulation of mitochondrial cell death protein BNIP3. Oncogene 24: 980-991.
Kerr JF, Wyllie AH, Currie AR. 1972. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26: 239-257.
Kharbanda S, Saxena S, Yoshida K, Pandey P, Kaneki M, Wang Q, et al. 2000. Translocation of SAPK/JNK to mitochondria and interaction with Bcl-x(L) in response to DNA damage. J Biol Chem 275: 322-327.
Kil WJ, Cerna D, Burgan WE, Beam K, Carter D, Steeg PS, et al. 2008. In vitro and in vivo radiosensitization induced by the DNA methylating agent temozolomide. Clin Cancer Res 14: 931-938.
Kim JH, Choi YW, Park C, Jin CY, Lee YJ, Park DJ, et al. 2009. Apoptosis induction of human leukemia U937 cells by gomisin N, a dibenzocyclooctadiene lignan, isolated from Schizandra chinensis Baill. Food Chem Toxicol.
Klionsky DJ, Emr SD. 2000. Autophagy as a regulated pathway of cellular degradation. Science 290: 1717-1721.
Ko H, Kim YJ, Park JS, Park JH, Yang HO. 2009. Autophagy inhibition enhances apoptosis induced by ginsenoside Rk1 in hepatocellular carcinoma cells. Biosci Biotechnol Biochem 73: 2183-2189.
Kondo Y, Kondo S. 2006. Autophagy and cancer therapy. Autophagy 2: 85-90.
Kurz EU, Lees-Miller SP. 2004. DNA damage-induced activation of ATM and ATM-dependent signaling pathways. DNA Repair (Amst) 3: 889-900.
Leist M, Jaattela M. 2001. Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2: 589-598.
Levine EL, Renehan A, Gossiel R, Davidson SE, Roberts SA, Chadwick C, et al. 1995. Apoptosis, intrinsic radiosensitivity and prediction of radiotherapy response in cervical carcinoma. Radiother Oncol 37: 1-9.
Li J, Qin Z, Liang Z. 2009. The prosurvival role of autophagy in Resveratrol-induced cytotoxicity in human U251 glioma cells. BMC Cancer 9: 215.
Li Y, Qu X, Qu J, Zhang Y, Liu J, Teng Y, et al. 2009. Arsenic trioxide induces apoptosis and G2/M phase arrest by inducing Cbl to inhibit PI3K/Akt signaling and thereby regulate p53 activation. Cancer Lett 284: 208-215.
Li YM, Broome JD. 1999. Arsenic Targets Tubulins to Induce Apoptosis in Myeloid Leukemia Cells. Cancer Res 59: 776-780.
Ling YH, Jiang JD, Holland JF, Perez-Soler R. 2002. Arsenic trioxide produces polymerization of microtubules and mitotic arrest before apoptosis in human tumor cell lines. Mol Pharmacol 62: 529-538.
Liu B, Cheng Y, Zhang B, Bian HJ, Bao JK. 2009. Polygonatum cyrtonema lectin induces apoptosis and autophagy in human melanoma A375 cells through a mitochondria-mediated ROS-p38-p53 pathway. Cancer Lett 275: 54-60.
Lockshin RA, Zakeri Z. 2001. Programmed cell death and apoptosis: origins of the theory. Nat Rev Mol Cell Biol 2: 545-550.
Lockshin RA, Zakeri Z. 2004. Apoptosis, autophagy, and more. Int J Biochem Cell Biol 36: 2405-2419.
Marina N, Gebhardt M, Teot L, Gorlick R, Marina N, Gebhardt M, et al. 2004. Biology and therapeutic advances for pediatric osteosarcoma. Oncologist 9: 422-441.
Marulanda GA, Henderson ER, Johnson DA, Letson GD, Cheong D, Marulanda GA, et al. 2008. Orthopedic surgery options for the treatment of primary osteosarcoma. Cancer Control 15: 13-20.
McLean K, VanDeVen NA, Sorenson DR, Daudi S, Liu JR. 2009. The HIV protease inhibitor saquinavir induces endoplasmic reticulum stress, autophagy, and apoptosis in ovarian cancer cells. Gynecol Oncol 112: 623-630.
McNair FI, Marples B, West CM, Moore JV. 1997. A comet assay of DNA damage and repair in K562 cells after photodynamic therapy using haematoporphyrin derivative, methylene blue and meso-tetrahydroxyphenylchlorin. Br J Cancer 75: 1721-1729.
Mervis J. 1996. Ancient remedy performs new tricks. Science 273: 578.
Miller WH, Jr., Schipper HM, Lee JS, Singer J, Waxman S, Miller WH, Jr., et al. 2002. Mechanisms of action of arsenic trioxide. Cancer Research 62: 3893-3903.
Mishra KP. 2004. Cell membrane oxidative damage induced by gamma-radiation and apoptotic sensitivity. J Environ Pathol Toxicol Oncol 23: 61-66.
Miyata H, Doki Y, Yamamoto H, Kishi K, Takemoto H, Fujiwara Y, et al. 2001. Overexpression of CDC25B overrides radiation-induced G2-M arrest and results in increased apoptosis in esophageal cancer cells. Cancer Research 61: 3188-3193.
Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, et al. 2001. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol 152: 657-668.
Mizushima N, Yoshimori T. 2007. How to interpret LC3 immunoblotting. Autophagy 3: 542-545.
Murray D, McEwan AJ. 2007. Radiobiology of systemic radiation therapy. Cancer Biother Radiopharm 22: 1-23.
Nakamura Y, Yogosawa S, Izutani Y, Watanabe H, Otsuji E, Sakai T. 2009. A combination of indol-3-carbinol and genistein synergistically induces apoptosis in human colon cancer HT-29 cells by inhibiting Akt phosphorylation and progression of autophagy. Mol Cancer 8: 100.
Ning S, Knox SJ. 2006. Optimization of combination therapy of arsenic trioxide and fractionated radiotherapy for malignant glioma. Int J Radiat Oncol Biol Phys 65: 493-498.
Olive PL. 1998. The role of DNA single- and double-strand breaks in cell killing by ionizing radiation. Radiat Res 150: S42-51.
Olive PL, Banath JP, Durand RE. 1990. Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the "comet" assay. Radiat Res 122: 86-94.
Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, et al. 2007. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 282: 24131-24145.
Park. J-W, Choi. Y-J, Jang. MA, Baek. S-H, Lim. JH, Passaniti. T, et al. 2001. Arsenic Trioxide Induces G2/M Growth Arrest and Apoptosis after Caspase-3 Activation and Bcl-2 Phosphorylation in Promonocytic U937 Cells. . Biochemical and Biophysical Research Communications 286: 726-734.
Peng PL, Kuo WH, Tseng HC, Chou FP. 2008. Synergistic tumor-killing effect of radiation and berberine combined treatment in lung cancer: the contribution of autophagic cell death. Int J Radiat Oncol Biol Phys 70: 529-542.
Pietenpol JA, Stewart ZA. 2002. Cell cycle checkpoint signaling: cell cycle arrest versus apoptosis. Toxicology 181-182: 475-481.
Poon RY, Chau MS, Yamashita K, Hunter T. 1997. The role of Cdc2 feedback loop control in the DNA damage checkpoint in mammalian cells. Cancer Res 57: 5168-5178.
Poruchynsky MS, Wang EE, Rudin CM, Blagosklonny MV, Fojo T. 1998. Bcl-xL is phosphorylated in malignant cells following microtubule disruption. Cancer Res 58: 3331-3338.
Pruschy M, Resch H, Shi YQ, Aalame N, Glanzmann C, Bodis S. 1999. Ceramide triggers p53-dependent apoptosis in genetically defined fibrosarcoma tumour cells. Br J Cancer 80: 693-698.
Qian W, Liu J, Jin J, Ni W, Xu W. 2007. Arsenic trioxide induces not only apoptosis but also autophagic cell death in leukemia cell lines via up-regulation of Beclin-1. Leuk Res 31: 329-339.
Rahmani M, Reese E, Dai Y, Bauer C, Payne SG, Dent P, et al. 2005. Coadministration of histone deacetylase inhibitors and perifosine synergistically induces apoptosis in human leukemia cells through Akt and ERK1/2 inactivation and the generation of ceramide and reactive oxygen species. Cancer Res 65: 2422-2432.
Roboz GJ, Dias S, Lam G, Lane WJ, Soignet SL, Warrell RP, Jr, et al. 2000. Arsenic trioxide induces dose- and time-dependent apoptosis of endothelium and may exert an antileukemic effect via inhibition of angiogenesis. Blood 96: 1525-1530.
Roh MS, Kim CW, Park BS, Kim GC, Jeong JH, Kwon HC, et al. 2004. Mechanism of histone deacetylase inhibitor Trichostatin A induced apoptosis in human osteosarcoma cells. Apoptosis 9: 583-589.
Ruvolo PP, Deng X, May WS. 2001. Phosphorylation of Bcl2 and regulation of apoptosis. Leukemia 15: 515-522.
Sausville EA. 2003. Cyclin-dependent kinase modulators studied at the NCI: pre-clinical and clinical studies. Curr Med Chem Anticancer Agents 3: 47-56.
Scarlatti F, Sala G, Ricci C, Maioli C, Milani F, Minella M, et al. 2007. Resveratrol sensitization of DU145 prostate cancer cells to ionizing radiation is associated to ceramide increase. Cancer Lett 253: 124-130.
Schmidt M, Bastians H, Schmidt M, Bastians H. 2007. Mitotic drug targets and the development of novel anti-mitotic anticancer drugs. Drug Resist Updat 10: 162-181.
Segers K, Backhovens H, Singh SK, De Voecht J, Ramael M, Van Broeckhoven C, et al. 1995. Immunoreactivity for p53 and mdm2 and the detection of p53 mutations in human malignant mesothelioma. Virchows Arch 427: 431-436.
Sentman CL, Shutter JR, Hockenbery D, Kanagawa O, Korsmeyer SJ. 1991. bcl-2 inhibits multiple forms of apoptosis but not negative selection in thymocytes. Cell 67: 879-888.
Shah MA, Schwartz GK. 2001. Cell cycle-mediated drug resistance: an emerging concept in cancer therapy. Clin Cancer Res 7: 2168-2181.
Shen JK, Du HP, Yang M, Wang YG, Jin J. 2009. Casticin induces leukemic cell death through apoptosis and mitotic catastrophe. Ann Hematol 88: 743-752.
Shen ZX, Chen GQ, Ni JH, Li XS, Xiong SM, Qiu QY, et al. 1997. Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients. Blood 89: 3354-3360.
Shim MJ, Kim HJ, Yang SJ, Lee IS, Choi HI, Kim T. 2002. Arsenic trioxide induces apoptosis in chronic myelogenous leukemia K562 cells: possible involvement of p38 MAP kinase. J Biochem Mol Biol 35: 377-383.
Smith ML, Fornace AJ, Jr. 1996. Mammalian DNA damage-inducible genes associated with growth arrest and apoptosis. Mutat Res 340: 109-124.
Smits VA, Medema RH. 2001. Checking out the G(2)/M transition. Biochim Biophys Acta 1519: 1-12.
Soignet SL, Frankel SR, Douer D, Tallman MS, Kantarjian H, Calleja E, et al. 2001. United States multicenter study of arsenic trioxide in relapsed acute promyelocytic leukemia. J Clin Oncol 19: 3852-3860.
Song KS, Kim JS, Yun EJ, Kim YR, Seo KS, Park JH, et al. 2008. Rottlerin induces autophagy and apoptotic cell death through a PKC-delta-independent pathway in HT1080 human fibrosarcoma cells: the protective role of autophagy in apoptosis. Autophagy 4: 650-658.
Tahir SK, Han EK, Credo B, Jae HS, Pietenpol JA, Scatena CD, et al. 2001. A-204197, a new tubulin-binding agent with antimitotic activity in tumor cell lines resistant to known microtubule inhibitors. Cancer Res 61: 5480-5485.
Tait JF. 2008. Imaging of apoptosis. J Nucl Med 49: 1573-1576.
Tallman MS. 2008. The expanding role of arsenic in acute promyelocytic leukemia. Semin Hematol 45: S25-29.
Tan ML, Ooi JP, Ismail N, Moad AI, Muhammad TS. 2009. Programmed cell death pathways and current antitumor targets. Pharm Res 26: 1547-1560.
Tang N, Song WX, Luo J, Haydon RC, He TC, Tang N, et al. 2008. Osteosarcoma development and stem cell differentiation. Clin Orthop 466: 2114-2130.
Tardito S, Bussolati O, Gaccioli F, Gatti R, Guizzardi S, Uggeri J, et al. 2006. Non-apoptotic programmed cell death induced by a copper(II) complex in human fibrosarcoma cells. Histochem Cell Biol 126: 473-482.
Toyoshima M. 2009. Analysis of p53 dependent damage response in sperm-irradiated mouse embryos. J Radiat Res (Tokyo) 50: 11-17.
Tsuchihara K, Fujii S, Esumi H. 2009. Autophagy and cancer: dynamism of the metabolism of tumor cells and tissues. Cancer Lett 278: 130-138.
Uckun FM, Stewart CF, Reaman G, Chelstrom LM, Jin J, Chandan-Langlie M, et al. 1995. In vitro and in vivo activity of topotecan against human B-lineage acute lymphoblastic leukemia cells. Blood 85: 2817-2828.
Ural AU, Avcu F, Candir M, Guden M, Ozcan MA. 2006. In vitro synergistic cytoreductive effects of zoledronic acid and radiation on breast cancer cells. Breast Cancer Res 8: R52.
Vaux DL, Aguila HL, Weissman IL. 1992. Bcl-2 prevents death of factor-deprived cells but fails to prevent apoptosis in targets of cell mediated killing. Int Immunol 4: 821-824.
Vogel C, Hager C, Bastians H. 2007. Mechanisms of mitotic cell death induced by chemotherapy-mediated G2 checkpoint abrogation. Cancer Res 67: 339-345.
Wada T, Penninger JM. 2004. Mitogen-activated protein kinases in apoptosis regulation. Oncogene 23: 2838-2849.
Wang D, Kreutzer DA, Essigmann JM. 1998. Mutagenicity and repair of oxidative DNA damage: insights from studies using defined lesions. Mutat Res 400: 99-115.
Wheeler JA, Stephens LC, Tornos C, Eifel PJ, Ang KK, Milas L, et al. 1995. ASTRO Research Fellowship: apoptosis as a predictor of tumor response to radiation in stage IB cervical carcinoma. American Society for Therapeutic Radiology and Oncology. Int J Radiat Oncol Biol Phys 32: 1487-1493.
Wittig JC, Bickels J, Priebat D, Jelinek J, Kellar-Graney K, Shmookler B, et al. 2002. Osteosarcoma: a multidisciplinary approach to diagnosis and treatment. Am Fam Physician 65: 1123-1132.
Xie L, Song X, Yu J, Wei L, Song B, Wang X, et al. 2009. Fractionated irradiation induced radio-resistant esophageal cancer EC109 cells seem to be more sensitive to chemotherapeutic drugs. J Exp Clin Cancer Res 28: 68.
Yamamura H, Yoshikawa H, Tatsuta M, Akedo H, Takahashi K. 1998. Expression of the smooth muscle calponin gene in human osteosarcoma and its possible association with prognosis. Int J Cancer 79: 245-250.
Yan CH, Liang ZQ, Gu ZL, Yang YP, Reid P, Qin ZH. 2006. Contributions of autophagic and apoptotic mechanisms to CrTX-induced death of K562 cells. Toxicon 47: 521-530.
Zhang Y, Wu Y, Wu D, Tashiro S, Onodera S, Ikejima T. 2009. NF-kappab facilitates oridonin-induced apoptosis and autophagy in HT1080 cells through a p53-mediated pathway. Arch Biochem Biophys 489: 25-33.
Zhao H, Guo W, Peng C, Ji T, Lu X. 2009. Arsenic trioxide inhibits the growth of Adriamycin resistant osteosarcoma cells through inducing apoptosis. Mol Biol Rep.
Zheng YT, Shahnazari S, Brech A, Lamark T, Johansen T, Brumell JH. 2009. The adaptor protein p62/SQSTM1 targets invading bacteria to the autophagy pathway. J Immunol 183: 5909-5916.


陳建安。淺談癌症的放射線治療。敏盛醫院放射腫瘤科。台灣。
安 威、李凌松。醫學細胞生物學。合計圖書出版社。台灣。
瞿中和、王喜忠、丁明孝。細胞分子生物學。九州圖書文物有限公司。台灣。
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