最近的研究指出，放射線能有效的治療癌症是由於它能藉由產生活性氧物種進而誘使癌細胞走向細胞凋亡。儘管，放射線能誘發細胞凋亡的發生，但它並不是對所有的癌症細胞都具有相同的治療效果，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。近來有證據指出三氧化二砷能增加癌細胞對三氧化二砷的敏感度。因此本實驗的目的是想探討在對放射線具有抗性的HL-60細胞中，三氧化二砷是否能增加放射線誘發細胞凋亡的能力。本實驗將利用Soft agar assay和flow cytometry來估量細胞經過處理後的存活情形及細胞是否進行細胞凋亡，再利用In Vitro Caspase like activity assay和Western blotting來分析細胞中訊號傳遞路徑改變情況。結果顯示三氧化二砷合併放謝線處理細胞後能減少細胞的存活率並且增加細胞凋亡的發生。在細胞中也能發現粒線體膜電位差的下降，Bax、PARP、cytochrome C、 caspase 3、9等蛋白被活化的現象。最重要的是合併處理放射線及三氧化二砷能增加細胞內活性氧物種的產生。細胞內活性氧物種在放射線合併三氧化二砷治療癌症的機制中似乎扮演著重要的角色。本實驗的結果指出三氧化二砷能幫助放射線的治療效果並且有希望能更廣泛的被應用在血癌的治療上。

Recent reports indicated a broad spectrum of anticancer activity of ionizing radiation (IR) due to its ability to induce apoptosis via intracellular production of intracellular reactive oxygen species (ROS). Despite its potent apoptotic effect, IR is not equally effective in all cancer cells, which has prompted a search for agents enhancing IR efficacy. Recently, evidence has been gathered that arsenic trioxide (As2O3) may sensitize tumor cells to IR treatment. The aim of our investigation was to evaluate whether As2O3enhances IR-mediated apoptosis in IR-resistant HL-60 cells. Soft agar assay and flow cytometry were used to assess cell viability and apoptosis. In Vitro Caspase like activity assay, and Western blotting were performed to analyze signaling alteration. Combined treatment with IR and As2O3 reduced cells viability with a concomitant increase of apoptosis. Apoptotic cell death was preceded by collapse of the mitochondrial membrane potential, increased expression of Bax, PARP, cytochrome C, caspase 3, caspase 9, activation. Importantly, the combined effect of IR and As2O3 was associated with increased production of intracellular ROS and was abolished by antioxidant N-Acetyl-Cysteine (NAC). Intracellular ROS as assumed as the key mediators contributing to the combined effect of IR and As2O3. These results indicate that As2O3 may facilitate the extention of therapeutic spectrum of IR and suggest that the combination of IR and As2O3 could be more broadly applied in leukemia therapy.

Akao Y. Mizoguchi H. Kojima S. Naoe T. Ohishi N. Yagi K. Arsenic induces apoptosis in B-cell leukaemic cell lines in vitro: activation of caspases and down-regulation of Bcl-2 protein. British Journal of Haematology. 102(4):1055-60, 1998 Sep.

An J. Kimura-Kuroda J. Hirabayashi Y. Yasui K. Development of a novel mouse model for dengue virus infection. Virology. 59(17):4369-74, 1999.

Bachleitner-Hofmann T. Kees M. Gisslinger H. Arsenic trioxide: acute promyelocytic leukemia and beyond. [Review] [34 refs] Review. Review, Tutorial] Leukemia & Lymphoma. 43(8):1535-40, 2002 Aug.

Bosma GC. Custer RP. Bosma MJ. A servere combined immunodeficiency mutation in the mouse. Nature. 301(5900):527-30, 1983.

Charley MR. Tharp M. Locker J. Deng JS. Goslen JB. Mauro T. McCoy P. Abell E. Jegasothy B. Establishment of a human cutaneous T-cell lymphoma in C.B-17 SCID mice. J Invest Dermatol. 94(3):381-384, 1990.

Chen Z. Brand NJ. Chen A. Chen SJ. Tong JH. Wang ZY. Waxman S. Zelent A. Fusion between a novel Kruppel-like zinc finger gene and the retinoic acid receptor-alpha locus due to a variant t(11;17) translocation associated with acute promyelocytic leukaemia. EMBO Journal. 12(3):1161-7, 1993 Mar.

Chhuttani PN. Chawla LS. Sharma TD. Arsenical neuropathy. Neurology. 17(3):269-74, 1967 Mar.

Chun YJ. Park IC. Park MJ. Woo SH. Hong SI. Chung HY. Kim TH. Lee YS. Rhee CH. Lee SJ. Enhancement of radiation response in human cervical cancer cells in vitro and in vivo by arsenic trioxide (As2O3). FEBS Letters. 519(1-3):195-200, 2002 May 22.

Csapo Z. Keszler G. Safrany G. Spasokoukotskaja T. Talianidis I. Staub M. Sasvari-Szekely M. Activation of deoxycytidine kinase by gamma-irradiation and inactivation by hyperosmotic shock in human lymphocytes. Biochemical Pharmacology. 65(12):2031-9, 2003 Jun 15.

Dai J. Weinberg RS. Waxman S. Jing Y. Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system. Blood. 93(1):268-77, 1999 Jan 1.

Davison K. Cote S. Mader S. Miller WH. Glutathione depletion overcomes resistance to arsenic trioxide in arsenic-resistant cell lines. Leukemia. 17(5):931-40, 2003 May.

Davison K. Mann KK. Waxman S. Miller WH Jr. JNK activation is a mediator of arsenic trioxide-induced apoptosis in acute promyelocytic leukemia cells. Blood. 103(9):3496-502, 2004 May 1.

Deng X. Hofmann ER. Villanueva A. Hobert O. Capodieci P. Veach DR. Yin X. Campodonico L. Glekas A. Cordon-Cardo C. Clarkson B. Bornmann WG. Fuks Z. Hengartner MO. Kolesnick R. Caenorhabditis elegans ABL-1 antagonizes p53-mediated germline apoptosis after ionizing irradiation. Nature Genetics. 36(8):906-12, 2004 Aug.

Dreher D. Junod AF. Role of oxygen free radicals in cancer development. European Journal of Cancer. 32( 1):30-8, 1996.

Estus S. Zaks WJ. Freeman RS. Gruda M. Bravo R. Johnson EM Jr. Altered gene expression in neurons during programmed cell death: identification of c-jun as necessary for neuronal apoptosis. Journal of Cell Biology. 127(6 Pt 1):1717-27, 1994 Dec.

Flores ER. Tsai KY. Crowley D. Sengupta S. Yang A. McKeon F. Jacks T. p63 and p73 are required for p53-dependent apoptosis in response to DNA damage. Nature. 416(6880):560-4, 2002 Apr 4.

Fuchs J. Hufljet M. Rothfuss L. Wilson DS. Carcamo G. Packer L. Impairment of enzymatic and non-enzymatic antioxidants in skin by UVB irradiation. J Invest Dermatol. 93:769–73, 1989.

Fulop GM. Phillips RA. The scid mutation in mice cause a general defect in DNA repair. Nature. 347(6292):479-482, 1990.

Glantz MJ. Burger PC. Friedman AH. Radtke RA. Massey EW. Schold SC Jr. Treatment of radiation-induced nervous system injury with heparin and warfarin.[see comment]. Neurology. 44(11):2020-7, 1994 Nov.

Gupta S. Yel L. Kim D. Kim C. Chiplunkar S. Gollapudi S. Arsenic trioxide induces apoptosis in peripheral blood T lymphocyte subsets by inducing oxidative stress: a role of Bcl-2. Molecular Cancer Therapeutics. 2(8):711-9, 2003 Aug.

Hengartner MO. The biochemistry of apoptosis.[see comment]. [Review] [75 refs] Nature. 407(6805):770-6, 2000 Oct 12.

Huang XJ. Wiernik PH. Klein RS. Gallagher RE. Arsenic trioxide induces apoptosis of myeloid leukemia cells by activation of caspases. Medical Oncology. 16(1):58-64, 1999 Apr.

Huang SY. Chang CS. Tang JL. Tien HF. Kuo TL. Huang SF. Yao YT. Chou WC. Chung CY. Wang CH. Shen MC. Chen YC. Acute and chronic arsenic poisoning associated with treatment of acute promyelocytic leukaemia. British Journal of Haematology. 103(4):1092-5, 1998 Dec.

Imahayashi S. Ichiyoshi Y. Yoshino I. Eifuku R. Takenoyama M. Yasumoto K. Tumor-infiltrating B-cell-derived IgG recognizes tumor components in human lung cancer. Cancer Invest. 18(6):530-536, 2000.

Jing Y. Dai J. Chalmers-Redman RM. Tatton WG. Waxman S. Arsenic trioxide selectively induces acute promyelocytic leukemia cell apoptosis via a hydrogen peroxide-dependent pathway. Blood. 94(6):2102-11, 1999 Sep 15.

Kaghad M. Bonnet H. Yang A. Creancier L. Biscan JC. Valent A. Minty A. Chalon P. Lelias JM. Dumont X. Ferrara P. McKeon F. Caput D. Monoallelically expressed gene related to p53 at 1p36, a region frequently deleted in neuroblastoma and other human cancers. Cell. 90(4):809-19, 1997.

Kawai K. Resetkova E. Enomoto T. Fornasier V. Volpe R. Is human leukocyte antigen-DR and intercellular adhesion molecule-1 expression on human thyrocytes constitutive in papillary thyroid cancer? Comparative studies in human thyroid xenografts in severe combined immunodeficient and nude mice. J. Clin. Endocrinol. Metab. 83(1):157-164, 1998.

Ko YC. A critical review of epidemiologic studies on black-foot disease. [Review] [10 refs] Journal of Uoeh. 8(3):339-53, 1986 Sep 1.

Kruszewski M. Wojewodzka M. Iwanenko T. Collins AR. Szumiel I. Application of the comet assay for monitoring DNA damage in workers exposed to chronic low-dose irradiation. II. Base damage. Mutation Research. 416(1-2):37-57, 1998 Aug 7.

Kurki S. Latonen L. Laiho M. Cellular stress and DNA damage invoke temporally distinct Mdm2, p53 and PML complexes and damage-specific nuclear relocalization. Journal of Cell Science. 116(Pt 19):3917-25, 2003 Oct 1.

Larochette N. Decaudin D. Jacotot E. Brenner C. Marzo I. Susin SA. Zamzami N. Xie Z. Reed J. Kroemer G. Arsenite induces apoptosis via a direct effect on the mitochondrial permeability transition pore. Experimental Cell Research. 249(2):413-21, 1999 Jun 15.

Lew YS. Kolozsvary A. Brown SL. Kim JH. Synergistic interaction with arsenic trioxide and fractionated radiation in locally advanced murine tumor. Cancer Research. 62(15):4202-5, 2002 Aug 1.

Li L. Story M. Legerski RJ. Cellular responses to ionizing radiation damage. International Journal of Radiation Oncology, Biology, Physics. 49(4):1157-62, 2001 Mar 15.

Li YM. Broome JD. Arsenic targets tubulins to induce apoptosis in myeloid leukemia cells. Cancer Research. 59(4):776-80, 1999 Feb 15.

Ling YH. Jiang JD. Holland JF. Perez-Soler R. Arsenic trioxide produces polymerization of microtubules and mitotic arrest before apoptosis in human tumor cell lines. Molecular Pharmacology. 62(3):529-38, 2002 Sep.

Lubaroff DM. Cohen MB. Schultz LD. Beamer WG. Survival of human prostate carcinoma, benign hyperplastic prostate tissues, and IL-2-activated lymphocytes in scid mice. Prostate. 27(1):32-41, 1995.

Ma BB. Bristow RG. Kim J. Siu LL. Combined-modality treatment of solid tumors using radiotherapy and molecular targeted agents. [Review] [209 refs] Journal of Clinical Oncology. 21(14):2760-76, 2003 Jul 15.

Martelli AM. Cappellini A. Tazzari PL. Billi AM. Tassi C. Ricci F. Fala F. Conte R. Casepase-9 is the upstream caspase activated by 8-methoxypsoralen and ultraviolet-A radiation treatment of Jurkat T leukemia cells and normal T lymphocytes. Haematologica. 89(4):471-9, 2004.

Miller WH Jr. Schipper HM. Lee JS. Singer J. Waxman S. Mechanisms of action of arsenic trioxide. [Review] [119 refs] Review. Review, Tutorial Cancer Research. 62(14):3893-903, 2002 Jul 15.

Montemaggi P. Costamagna G. Dobelbower RR. Cellini N. Morganti AG. Mutignani M. Perri V. Brizi G. Marano P. Intraluminal brachytherapy in the treatment of pancreas and bile duct carcinoma. International Journal of Radiation Oncology, Biology, Physics. 32(2):437-43, 1995 May 15.

Namgung U. Xia Z. Arsenite-induced apoptosis in cortical neurons is mediated by c-Jun N-terminal protein kinase 3 and p38 mitogen-activated protein kinase. Journal of Neuroscience. 20(17):6442-51, 2000 Sep 1.

Nathan CF. Root RK. Hydrogen peroxide release from mouse peritoneal macrophages: dependence on sequential activation and triggering. Journal of Experimental Medicine. 146(6):1648-62 1977.

Roboz GJ. Dias S. Lam G. Lane WJ. Soignet SL. Warrell RP Jr. Rafii S. Arsenic trioxide induces dose- and time-dependent apoptosis of endothelium and may exert an antileukemic effect via inhibition of angiogenesis. Blood. 96(4):1525-30, 2000 Aug 15.

Rojewski MT. Korper S. Thiel E. Schrezenmeier H. Depolarization of mitochondria and activation of caspases are common features of arsenic(III)-induced apoptosis in myelogenic and lymphatic cell lines. Chemical Research in Toxicology. 17(1):119-28, 2004 Jan.

Sakakibara T. Xu Y, Bumpers HL. Chen FA. Bankert RB. Arredondo MA. Edge SB. Repasky EA. Growth and Metastasis of Surgical Specimens of Human Breast Carcinomas in SCID Mice. Cancer J. Scl. Am. 2(5):291-300, 1996.

Shivelman E. Namikawa R. Species-specific metastasis of human tumor cells in the severe combined immunodeficiency mouse engrafted with human tissue. Proc. Natl. Acad. Sci. U.S.A. 92(10):4661-4665, 1995.

Smith AH. Arroyo AP. Mazumder DN. Kosnett MJ. Hernandez AL. Beeris M. Smith MM. Moore LE. Arsenic-induced skin lesions among Atacameno people in Northern Chile despite good nutrition and centuries of exposure. Environmental Health Perspectives. 108(7):617-20, 2000 Jul.

Soignet SL. Frankel SR. Douer D. Tallman MS. Kantarjian H. Calleja E. Stone RM. Kalaycio M. Scheinberg DA. Steinherz P. Sievers EL. Coutre S. Dahlberg S. Ellison R. Warrell RP Jr. United States multicenter study of arsenic trioxide in relapsed acute promyelocytic leukemia. [Clinical Trial. Journal Article. Multicenter Study] Journal of Clinical Oncology. 19(18):3852-60, 2001 Sep 15.

Stadtman ER. Berlett BS. Fenton chemistry. Amino acid oxidation. Journal of Biological Chemistry. 266(26): 17201-11, 1991.

Stern JE. Li Y. Zhang W. Nitric oxide: a local signalling molecule controlling the activity of pre-autonomic neurones in the paraventricular nucleus of the hypothalamus. Acta Physiologica Scandinavica..177(1):37-42, 2003.

Sturlan S. Baumgartner M. Roth E. Bachleitner-Hofmann T. Docosahexaenoic acid enhances arsenic trioxide-mediated apoptosis in arsenic trioxide-resistant HL-60 cells. Blood. 101(12):4990-7, 2003 Jun 15.

Suzuki YJ. Forman HJ. Sevanian A. Oxidants as stimulators of signal transduction. Free Radical Biology & Medicine. 22(1-2): 269-85, 1997.

Tanaka-Kagawa T. Hanioka N. Yoshida H. Jinno H. Ando M. Arsenite and arsenate activate extracellular signal-regulated kinases 1/2 by an epidermal growth factor receptor-mediated pathway in normal human keratinocytes. British Journal of Dermatology. 149(6):1116-27, 2003 Dec.

Urashima M. Chen BP. Chen S. Pinkus GS. Bronson RT. Dedera DA. Hoshi Y. Teoh G. Ogata A. Treon SP. Chauhan D. Anderson KC. The development of a model for the homing of multiple myeloma cells to human bone marrow. Blood. 90(2):754-765, 1997.

Wells G. Koh WJ. Pelton J. Russell K. Griffin B. Laramore G. Griffin T. Parker R. Peters LJ. Davis L. Fast neutron teletherapy in advanced epidermoid head and neck cancer. A review. American Journal of Clinical Oncology. 12(4):295-300, 1989 Aug.

Winter WE. Maxwell GL. Tian C. Sobel E. Rose GS. Thomas G. Carlson JW. Association of hemoglobin level with survival in cervical carcinoma patients treated with concurrent cisplatin and radiotherapy: a Gynecologic Oncology Group Study. Gynecologic Oncology. 94(2):495-501, 2004 Aug.

Wu Q. Kirschmeier P. Hockenberry T. Yang TY. Brassard DL. Wang L. McClanahan T. Black S. Rizzi G. Transcriptional regulation during
p21/WAF1/Cip1-induced apoptosis in human ovarian cancer cells. J.
Biol. Chem. 277, 36329–36337.

Yi J. Yang J. He R. Gao F. Sang H. Tang X. Ye RD. Emodin enhances arsenic trioxide-induced apoptosis via generation of reactive oxygen species and inhibition of survival signaling. Cancer Research. 64(1):108-16, 2004 Jan 1.

Zaika A. Irwin M. Sansome C. Moll UM. Oncogenes induce and activate endogenous p73 protein. Journal of Biological Chemistry. 276(14):11310-6, 2001 Apr 6.

Zheng Y. Shi Y. Tian C. Jiang C. Jin H. Chen J. Almasan A. Tang H. Chen Q. Essential role of the voltage-dependent anion channel (VDAC) in mitochondrial permeability transition pore opening and cytochrome c release induced by arsenic trioxide. Oncogene. 23(6):1239-47, 2004 Feb 12.

Zhu J. Koken MH. Quignon F. Chelbi-Alix MK. Degos L. Wang ZY. Chen Z. de The H. Arsenic-induced PML targeting onto nuclear bodies: implications for the treatment of acute promyelocytic leukemia. Proceedings of the National Academy of Sciences of the United States of America. 94(8):3978-83, 1997 Apr 15.