臺灣博碩士論文加值系統

樟芝(學名Antrodia cinnamomea)，是一種生長在牛樟樹上的蕈類，且為台灣特有的傳統中藥。先前的研究發現樟芝發酵液對雌激素依賴性乳癌細胞株MCF-7具有生長抑制且能誘導之細胞凋亡的作用，因此根據這些數據，我們將在本研究中探討有關樟芝發酵液(AC-10)對多種乳癌細胞株之細胞毒性及其可能作用之機制。首先我們先檢測各種乳癌細胞株(MCF-7, MDA-MB-231, AU565 and BT474)之細胞存活率，結果發現以樟芝發酵液(AC-10)處理之乳癌細胞MCF-7與MDA-MB-231具有很強之細胞毒性，兩者之存活率都隨著AC-10作用的時間和濃度增加而下降，尤其是MDA-MB-231，其IC50 值約為136μg/ml，毒性較其他乳癌細胞株強，因此探討其可能的機制。為了確認AC-10對MDA-MB-231之影響，利用倒立式螢光顯微鏡、單細胞凝膠電泳法(Comet assay)及DNA梯狀圖譜進行分析，結果顯示AC-10可以誘導MDA-MB-231細胞凋亡，其特徵包括細胞皺縮(cell shrinkage)、DNA之損傷及斷裂(DNA fragmentation)，且隨著AC-10處理濃度及作用時間增加而增加。之後，我們又以西方點墨法(Western blotting)的方式偵測AC-10對MDA-MB-231細胞之細胞凋亡相關蛋白的影響，結果發現AC-10對MDA-MB-231誘導之細胞凋亡機制，會伴隨著細胞質中cytochrome c的增加，Bax/Bcl-2比率之上升，以及 procaspase-8, procaspase-9, procaspase-3 and poly(ADP)- ribosylpolymerase (PARP)之斷裂。另外，研究也發現在AC-10處理後之環氧化酶-2(cyclooxygenase-2；COX-2)蛋白表現及其產物PGE2 的生成都顯著的減少。然而，從西方點墨法與電泳遷移率變動實驗（Electrophoretic mobility shift assay， EMSA）中得知，AC-10會誘使MDA-MB-231細胞質中IκB-α 蛋白被降解進而導致 NF-κB 的持續性活化而時間可長達24小時。因此，這些數據說明樟之發酵液(AC-10)對MDA-MB-231乳癌細胞株之細胞毒性的機制，可能是以誘導其細胞凋亡的路徑所致，而研究中發現NF-κB長時間持續性活化可能會導致AC-10調控MDA-MB-231細胞以其他路徑調控細胞凋亡。最後本實驗提供的證據顯示，樟芝發酵液(AC-10) 可能具有其潛力發展成抑制乳癌細胞生長之藥劑的能力。

Antrodia cinnamomea (A. cinnamomea) is well known in Taiwan as a traditional Chinese medicine, and has been reported growth inhibition on estrogen-positive MCF-7 breast cancer cells through apoptosis induction. According to the previous data, we also analyzed its cytotoxic effect on estrogen-negative MDA-MB-231 breast cancer cells and underlying mechanisms. First, we investigated the effect of the fermented culture broth of A. cinnamomea (AC-10) on cell viability of breast cancer cell lines (MCF-7, MDA-MB-231, AU565 and BT474), and the induction of apoptosis in MDA-MB-231 cells. After 24 hours of treatment with AC-10, both of dose- and time dependent decreases in the viability of MCF-7 and MDA-MB-231 cells were observed. The cytotoxic effect of AC-10 on MCF-7 cells was less significant than that on MDA-MB-231 cells after 24 hours of treatment. In order to confirm the effect of AC-10 on MDA-MB-231 cells, we analyzed DNA laddering, Comet assay and western blotting of apoptotic related protein. Induction of apoptosis in AC- treated MDA-MB-231 cells was accompanied by an increase of cytosolic cytochrome c, an up-regulation of Bax/Bcl-2 ratio, and cleavages of procaspase-8, procaspase-9, procaspase-3 and poly(ADP)- ribosylpolymerase (PARP). After treatment of AC-10 in MDA-MB-231, protein expression of COX-2(cyclooxygenase-2；COX-2) and its products PGE2 were markedly reduced. Furthermore, IκB-α degradation and NF-κB activity of MDA-MB-231 cells induced by AC-10 were activated continuously for 24 hours . Therefore, these results suggested that fermented culture broth of A. cinnamomea (AC-10) may exert antiproliferative action and induce apoptosis on MDA-MB-231 breast cancer cells through another mechanism which may relate to the lasting activated effect of NF-κB. And this study provide evidences that A. cinnamomea may be a potential anti-cancer agent for the treatment of breast cancer.

中文摘要--------------------------------1
英文摘要--------------------------------3
第一章、前言----------------------------7
第一節、研究背景----------------------8
圖一---------------------------10
表一---------------------------12
表二---------------------------14
圖二---------------------------20
表三---------------------------24
圖三---------------------------25
表四---------------------------26
圖四---------------------------28
圖五---------------------------33
圖六---------------------------34
圖七、八------------------------36
第二節、研究動機----------------------38
弟二章、研究方法------------------------39
第一節、研究器材----------------------40
第二節、研究設計與方法----------------43
第三節、實驗方法----------------------45
第三章、研究結果與圖表------------------56
Figure 1-1 ----------------------63
Figure 1-2 ----------------------64
Figure 2 ------------------------65
Figure 3-1 ----------------------67
Figure 3-2 ----------------------68
Figure 3-3 ----------------------69
Figure 4-1 ----------------------72
Figure 4-2 ----------------------73
Figure 5-1 ----------------------75
Figure 5-2 ----------------------76
Summary-1 -----------------------77
Figure 6-1 ----------------------79
Figure 6-2 ----------------------80
Figure 7 ------------------------82
Figure 8 ------------------------84
Summary-2 -----------------------85
第四章、討論----------------------------86
第五章、結論----------------------------91
第六章、參考文獻------------------------93

1.陳勁初以及呂鋒洲. 靈芝之王:臺灣樟芝, 2001.
2.Zang M. and Su C.H. Ganoderma comphoratum, a new taxon in genus Ganoderma From Taiwan, China. Acta Botanical Yunnanical,12: 395-396, 1990.
3.Chang T. T. and Wang W.R. Basidiomal formation of Antrodia cinnamomea on artificial agar media. Bot. Bull. Acad. Sin. 46：151-154, 2005.
4.Chang T. T. and Chou W. N. Antrodia cinnamomea reconsidered and A. salmonea sp. nov. on Cunninghamia konishii in Taiwan.Bot. Bull. Acad. Sin. 45:347-352, 2004.
5.陳勁初、林文鑫、陳清農、許勝傑、黃仕政以及陳炎鍊.台灣特有真菌-樟芝菌絲體之開發. Fungal Science 16卷: 7-22, 2001
6. 張東柱. 牛樟之病害. 牛樟生物學及育林技術研討會論文集. 林業叢刊 第72號:127-131, 1997.
7. 許勝傑、陳清農以及陳勁初. 樟芝宿主專一性之探討. 台灣農業化學與食品科學 38: 533-539, 2000.
8. 黃鈴娟. 樟芝與姬松茸之抗氧化性質及其多醣組成分析. 國立中興大學 食品科學系, 2000.
9. Chang H. L., Chao G. R., Chen C. C. and Mau J. L. Non-volatile taste components of Agaricus blazei, Antrodia camphorata and Cordyceps militaris mycelia. Food Chem. 74: 203-207, 2001.
10. Hirotani M. and Furuya T. Changes of the triterpenoid patterns during formation of the fruit body in Ganoderma. Phytochem. 29: 3767-37711, 1990.
11. Su C. H., Yang Y. Z., Ho H. O., Hu C. H. and Sheu M. T. High- performance liquid chromatographic analysis for the characterization of triterpenoids from Ganoderma. J. Chromato. Sci. 39: 93-100, 2001.
12. Kim H. W. and Kim B. K. Biomedicinal Triterpenoids of Ganoderma lucidum (Aphyllophoromycetideae). Inter. J. Med. Mushrooms 1: 121-138, 1999.
13. Toth J. O., Luu B. and Ourisson G. Cytotoxic triterpenes from Ganoderma lucidum. Tetrahedron Lett. 24: 1081-1084, 1983.
14. Gan K. H., Fann Y. F., Hsu S. H., Kuo K. W. and Lin C. N. Mediation of the cytotoxicity of lanostanoids and steroids of Ganoderma tsugae through apoptosis and cell cycle. J. Nat. Prod. 61: 485-487, 1998.
15. Wang S. Y., Hsu M. L., Hsu H. C., Tzeng C. H., Lee S. S., Shiao M. S. and Ho C. K. The anti-tumor effect of Ganoderma lucidum is mediated by cytokines released from activated macrophages. Int. J. Cancer 70: 699-705, 1997.
16. Kohda H., Tokumoto W., Sakamoto K., Fujii M., Hirai Y., Yamasaki K., Komoda Y., Nakamura H., Ishihara S. and Uchida M. The biologically active constituents of Ganoderma lucidum (FR.) KARST. Histamine release- inhibitory Triterpenes. Chem. Pharm. Bull. 33: 1367-1374, 1985.
17. Chen D. H., Shiou W. Y., Wang K. C., Huang S. Y., Shie Y. T., Tsai C. M., Shi, J. F. and Chen K. D. Chemotaxonomy of triterpenoid pattern of HPLC of Ganoderma lucidum and Ganoderma tsugae. J. Chinese Chem. Soc. 46: 47-51, 1999.
18. Cherng I. H. and Chiang H. C. Three new triterpenoids from Antrodia cinnamomea. J. Natural Products 58: 365-371, 1995.
19. Cherng I. H., Wu D. P. and Chiang H. C. Triterpenoids from Antrodia cinnamomea. Phytochemistry 41: 263-267, 1996.
20. 吳德鵬. 樟芝微量成分的研究. 國立台灣師範大學化學研究所碩士論文. 1995
21. 簡秋源、姜宏哲以及陳淑貞. 牛樟菇培養性狀及其三帖類成分分析之研究，牛樟生物學及育林技術研討會論文集. 林業叢刊第72號：133-137, 1997.
22. Chen C. H. and Yang S. W. New steroid acids from Antrodia cinnamomea a fungal parasite of Cinnamomum micranthum. J. Natural Products 58: 1655-1661. 1995.
23. Sone Y., Okuda R., Wada N., Kishida E. and Misaki A. Structures and antitumor activaties of the polysaccharides isolated from fruiting body and the growing culture of mycelium of Ganoderma lucidum. Agric. Biol. Chem. 49: 2641-53, 1985.
24. Wang G., Zhang J., Mizuno, T., Zhuang C., Ito H., Mayuzumi H., Okamoto H. and Li J. Antitumor active polysaccharides from the Chinese mushroom Songshan Lingzhi, the fruiting body of Ganoderma tsugae. Biosci. Biotech. Biochem. 57: 894-900, 1993.
25. Wasser S. P. and Weis A. L. Medicinal properties of substances occurring in higher basidomycetes mushrooms: current perspectives (review). Intern. J. Med. Mushrooms. 1:31-62, 1999.
26. Wasser S. P. and Weis A. L. Therapeutic effects of substances occurring in higher basidiomycetes mushrooms: a modern perspective. Critical ReviewsTM in Immunology. 19: 65-96, 1999.
27. Wang S. Y., Hsu M. L., Hsu H. C., Tzeng C. H., Lee S. S., Shiao M. S. and Ho C. K. The anti-tumor effect of Ganoderma lucidum is mediated by cytokines released from activated macrophages. Int. J. Cancer 70: 699-705, 1997.
28. Eo S. K., Kim Y. S., Lee C. K. and Han S. S. Antiherpetic activities of various protein bound polysaccharide isolated from Ganoderma lucidum. J Ethnopharma. 72: 475-481, 1999.
29. Mizuno T. The extraction and development of antitumor-active polysaccharides from medicinal mushrooms in Japan (Review). Inter. J. Med. Mushrooms. 1: 9-29, 1999.
30. Lee I. H., Huang R. L., Chen C. T., Chen H. C., Hsu W. C. and Lu M. K. Antrodia camphorate polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiol. Letters 209: 63-67, 2002.
31. 江素瑛、吳焜裕、王秋萍、魏逸傑、施睿宏、高尚德以及謝慶良. 樟芝之致基因毒性與抗基因毒性評估. 台灣保健食品協會 第二屆第二次年會 大會議程及論文摘要. p.48, 2002.
32. 郭淑卿. 2002. 樟芝發酵液對大鼠肝臟纖維化及胃腸功能之
改善作用. 中國醫藥學院 中國藥學研究所藥學碩士論文.
33. 林文鑫、陳俊憲、陳勁初以及呂鋒州. 樟芝液態發酵萃取物對腫瘤細胞株之毒殺性分析. 中華民國食品科學技術學會 第三十次(第十五屆第二次)會員大會手冊 p. 254, 2000.
34. 江素瑛. 靈芝與樟芝之安全與毒理. 「保健食品」跨部會整合推動委員會 p. 64-72, 2001.
35. 沈立言. 牛樟芝菌絲體發酵萃取液對肝臟生理機能性之影響. 89年度保健食品研究開發計畫心得發表會 p.191-198, 2000.
36. Huang L. C., Huang S. J., Chen C. C. and Mau J. L. Antioxidant properties of Antrodia camphorata. Proceedings of 3rd International Conference on Mushroom Biology & Mushroom Products. p275-283, 1999.
37. Hseu Y. C., Chang W. C., Hseu Y. T., Lee C. Y. Yech Y. J., Chen P. C., Chen J. Y. and Yang H. L. Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sci. 71: 469-482, 2002.
38. 劉翠玲. 樟芝對倉鼠體內脂質代謝與抗氧化狀態之影響. 輔仁大學 食品營養學系碩士論文, 2002.
39. 戴宇昀. 樟芝菌絲體與子實體對四氯化碳及酒精誘導之慢性及急性肝損傷之保肝功能評估. 國立中興大學 食品科學系研究所碩士論文, 2001.
40. 李炫璋、莊正宏、蔡金川、黃仕政、陳勁初以及胡淼琳. 中華民國食品科學技術學會. 第三十二次(第十六屆第二次)會員大會手冊 p. 339, 2002.
41. 陳怡欣. 牛樟芝發酵過濾液對大白鼠肝臟生理機能之影響. 中國醫藥學院 營養研究所碩士論文, 2002.
42. 陳秀雯、陳清農、黃仕政、許勝傑、曾虹萍、洪鳴遠、傅麒玲以及陳勁初. 樟芝菌絲體之免疫調節功能評估. 中華民國食品科學技術學會. 第三十二次(第十六屆第二次)會員大會手冊 p. 326, 2002.
43. 賴宏亮、呂美津、陳武元以及莊秀琪. 樟芝免疫調節活性之研究. 台灣保健食品學會. 第二屆第二次年會. 大會議程及壁報論文摘要. p. 49, 2002 .
44. 張中姿、陳俊憲、林文鑫、陳勁初以及呂鋒洲. 中華民國食品科學技術學會. 第三十二次(第十六屆第一次)會員大會手冊 p. 265, 2001.
45. 呂鋒洲. 牛樟芝各抗腫瘤有效成分的分離、純化，誘發腫瘤凋亡及其作用機轉的研究. 九十一年度保健食品研究開發計畫成果發表會. p.13-16, 2002.
46. 林文川. 樟芝之藥理學研究. 九十年度保健食品研究開發計畫成果摘要. p.55-57, 2001.
47. 中華民國 行政院衛生署之2004年台灣女性癌症死亡率統計資料。資料來源http://www.doh.gov.tw/statistic/index.htm
48.Hseu Y.C., Wu F.Y., Wu J.J., Chen J.Y., Chang W.H., Lu F.J., Lai Y.C. and Yang H.L. Anti-inflammatory potential of Antrodia Camphorata through inhibition of iNOS, COX-2 and cytokines via the NF-kappaB pathway.Int Immunopharmacol. 5 (13-14): 1914-25, 2005.
49.Hseu Y.C., Yang H.L., Lai Y.C., Lin J.G., Chen G.W. and Chang Y.H. Induction of apoptosis by Antrodia camphorata in human premyelocytic leukemia HL-60 cells. Nutr Cancer. 48(2):189-97, 2004.
50. Yang H.L., Chen C.S., Chang W.H., Lu F.J., Lai Y.C., Chen C.C., Hseu T.H., Kuo C.T. and Hseu Y.C. Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by Antrodia camphorata. Cancer Lett. 231(2):215-27, 2006.
51. Yang H.L., Hseu Y.C., Chen J.Y., Yech Y.J., Lu F.J., Wang H.H., Lin P.S. and Wang B.C. Antrodia camphorata in submerged culture protects low density lipoproteins against oxidative modification.Am J Chin Med. 34(2):217-31, 2006.
52.Robert E. Coleman Current and Future Status of Adjuvant Therapy for Breast cancer. Cancer 97: 880-886, 2003.
53. Intra M., Gatti G. and Luini A. Surgical technique of intraoperative radiotherapy in conservative treatment of limited-stage breast cancer.Arch Surg 137: 737-740, 2002.
54. Coates A.S., Gelber R.D. and Goldhirsch A. Subsets within the chemotherapy overview: International Breast Cancer Study Group. Lancet 352: 1783-1784, 1998.
55. Coates A.S., Goldhirsch A. and Gelber R.D. Overhauling the breast cancer overview: Are subsets subversive? Lancet Oncol 3: 525-526, 2002.
56. Bartelink H. Radiotherapy to the conserved breast, chest wall, and regional nodes: Is there a standard? Breast 12: 475-482, 2003.
57. Beatson C.T. On treatment of inoperable cases of carcinoma of the mamma: suggestions for a new method of treatment with illustrative cases. Lancet 2:104–7, 1896
58. Hayashi A., Silver S., van der Westhuizen N., Donald J., Parker C. and Fraser S. Treatment of invasive breast carcinoma with ultrasound guided radiofrequency ablation. Am J Surg 185(5):429-435, 2003.
59. Kuiper G.G.J.M., Enmark E., Pelto-Hukko M., Nilsson S.and Gustafsson J.A. Cloning of a novel estrogen receptor expressed in rat prostate and ovary. PNAS 93 : 5925-5930, 1996.
60. Omoto Y., Kobayashi S., Inoue S., Ogawa S., Toyama T., Yamashita H., Muramatsu M., Gustafsson J.A. and Iwase H. Evaluation of oestrogen receptor β wild-type and variant protein expression, and relationship with clinicopathological factors in breast cancers. Eur J Cancer 38 : 380-386, 2002.
61. Omoto Y., Kobayashi Y., Nishida K., Tsuchiya E., Eguchi H., Nakagawa K., Ishikawa Y., Yamori T., Iwase H., Fujii Y., Warner M., Gustafsson J.A. and Hayashi S.I. Expression, function and clinical implications of estrogen receptor β in human lung cancer. Biochem Biophys Res Commun 285 : 340-347, 2001.
62. Matsuyama S., Ohkura Y., Eguchi H., Kobayashi Y., Akagi K., Uchida K., Nakachi K., Gustafsson J.A. and Hayashi S.I. Estrogen receptor β (ERβ) is expressed in human stomach adenocarcinomas. J Cancer Res Clin Oncol 128: 319-324, 2002.
63. Fisher B., Costantino J.P., Wickerham D.L., Redmond C.K., Kavanah M., Cronin W.M., Vogel V., Robidoux A., Dimitrov N., Atkins J., Daly M., Wieand S., Tan-Chiu E., Ford L. and Wolmark N. Tamoxifen for prevention of breast cancer: Report of the National Surgical Adjuvant Breast and Bowel Project P-1 Study. J Natl Cancer Inst 90: 1371-1388, 1998.
64. Love R.R., Duc N.B., Allred D.C., Binh N.C., Dinh N.V., Kha N.N., Thuan T.V., Mohsin S.K., Roanh le D., Khang H.X., Tran T.L., Quy T.T., Thuy N.V., The P.N., Cau T.T., Tung N.D., Huong D.T., Quang le M., Hien N.N., Thuong L., Shen T.Z., Xin Y., Zhang Q., Havighurst T.C., Yang Y.F., Hillner B.E. and DeMets D.L. Oophorectomy and tamoxifen adjuvant therapy in premenopausal Vietnamese and Chinese women with operable breast cancer. J Clin Oncol 20: 2559-2566, 2002.
65. Cuzick J., Forbes J. and Howell A. Tamoxifen for breast-cancer prevention. Lancet 361:177-178, 2003.
66. Winer E.P., Hudis C., Burstein H.J., Chlebowski R.T., Ingle J.N., Edge S.B., Mamounas E.P., Gralow J., Goldstein L.J., Pritchard K.I., Braun S., Cobleigh M.A., Langer A.S., Perotti J., Powles T.J., Whelan T.J. and Browman G.P. American Society of Clinical Oncology technology assessment on the use of aromatase inhibitors as adjuvant therapy for women with hormone receptor-positive breast cancer: Status report. J Clin Oncol 20:3317-3327, 2002.
67. Loprinzi C.L., Michalak J.C., Quella S.K., O''Fallon J.R., Hatfield A.K., Nelimark R.A., Dose A.M., Fischer T., Johnson C., KlattN.E., Bate W.W., Rospond R.M. and Oesterling J.E. Megestrol acetate for the prevention of hot .ashes. N Engl J Med 331: 347-352, 1994.
68. Quella S.K., Loprinzi C.L., Sloan J.A., Vaught N.L., DeKrey W.L., Fischer T., Finck G., Pierson N. and Pisansky T. Long term use of megestrol acetate for the treatment of hot flashes in cancer survivors. Cancer; 82:1784-1788, 1998.
69. Jonat W., Kaufmann M. and Sauerbrei W. Goserelin versus cyclophosphamide, methotrexate, and fluorouracil as adjuvant therapy in premenopausal patients with node-positive breast cancer: The Zoladex Early Breast Cancer Research Association Study. J Clin Oncol 20: 4628-4635, 2002.
70. Greenspan E.M. Combination cytotoxic chemotherapy in advanced disseminated breast carcinoma. J Mt Sinai Hosp NY: 1-27, 1965.
71. Schiff P.B., Fant J. and Horwitz S.B.Promotion of microtubule assembly in vitro by Taxol. Nature 277: 665-667, 1979.
72.Leist M. and Nicotera, P. Apoptosis, excitotoxicity, and neuropathology. Exp.Cell Res. 239 (2): 183-201, 1998.
73. Shivapurkar N., Reddy J., Chaudhary P.M., Gazdar A.F. Apoptosis and lung cancer: A review. Journal of Cellular Biochemistry, 88:885–898, 2003.
74. Kerr J.F., Wyllie A.H. and Currie A.R. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26: 239-257, 1972.
75. Zimmermann K.C., Bonzon C. and Green D.R. The machinery of programmed cell death. Pharmacol Ther. 92: 57–70, 2001.
76. Wyllie A. H. Glucocorticoid-induced thmocyte apoptosis is associatedwith endogenous endonuclease activiation. Nature 284: 555-556, 1980.
77. Staunton M.J. and Gaffney E.F. Apoptosis: basic concepts and potential significance in human cancer. Archives of pathology & laboratory medicine. 122(4):310-9, 1998.
78. Alnemri, E.S., Livingston, D.J., Nicholson, D.W., Salvesen, G., Thornberry, N.A., Wong, W.W. and Yuan, J. Human ICE/CED-3 protease nomenclature. Cell 87: 171, 1996.
79. Nicholson D.W, Ali A., Thornberry N.A., Vaillancourt J.P., Ding C.K., Gallant M., Gareau Y., Griffin P.R., Labelle M. and Lazebnik Y.A. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 376: 37-43, 1995.
80. Bortner C.D. and Cidlowski J.A. Cellular mechanisms for the repression of apoptosis. Annu Rev Pharmacol Toxicol 42: 259-281, 2002.
81. Nicholson D. W. and Thornberry N. A. Apoptosis. Life and death decisions. Science: 299-306, 1997.
82. Wolf B.B. and Green D.R. Suicidal tendencies: apoptotic cell death by caspase family proteinases. J Biol Chem 274: 20049-20052, 1999.
83. Stennicke H.R. and Salvesen G.S. Properties of the caspases. Biochim Biophys Acta 1387: 17-31 1998.
84. Earnshaw W.C., Martins L.M. and Kaufmann S.H. Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem 68: 383-424, 1999.
85. Enari M., Sakahira H., Yokoyama H., Okawa K., Iwamatsu A. and Nagata S. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43-50, 1998.
86. Kothakota S., Azuma T., Reinhard C., Klippel A., Tang J., Chu K., McGarry T.J., Kirschner M.W., Koths K., Kwiatkowski D.J. and Williams L.T. Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis. Science 278: 294-298, 1997.
87. Rudel T. and Bokoch G.M. Membrane and morphological changes in apoptotic cells regulated by caspase-mediated activation of PAK2. Science 276: 1571-1574, 1997.
88. Rao L., Perez D. and White E. Lamin proteolysis facilitates nuclear events during apoptosis. J Cell Biol 135: 1441-1455, 1996.
89. Yang J., Liu X., Bhalla K., Kim C.N., Ibrado A.M., Cai J., Peng T.I., Jones D.P. and Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 275:1129-1132, 1997.
90. Li H., Zhu H., Xu C. and Yuan. J. Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94: 491-501, 1998.
91. Lou X., Budihardjo I., Zou H.,Slaughter C. and Wang X . Bid.a Bcl-2 interacting protein. Mediates cytochrom c release from mitochondria in response to activation of cell surface death receptor. Cell 94: 481-490, 1998.
92. Gibbons G.H. and Pollman M.J. Death receptors, intimal disease, and gene therapy: are therapies that modify cell fate moving too Fas? Circ Res 86: 1009-1012, 2000.
93. Ashkenazi A. and Dixit V.M. Death receptors: signaling and modulation. Science 281: 1305-1308, 1998.
94. Eskes R., Desagher S., Antonsson B. and Martinou J.C. Bid induces the oligomerization and insertion of Bax into the outer mitochondrial membrane. Mol Cell Biol 20: 929-935, 2000.
95. Nakagawa T., Zhu H., Morishima N., Li E., Xu J., Yankner B.A. and Yuan J. Caspase-12 mediates endoplasmic –reticulum -specific apoptosis and cytotoxicity by amyloid-beta. Nature, 403: 98-103, 2000.
96. Ferrari D., Pinton P., Szabadkai G., Chami M., Campanella M., Pozzan T. and Rizzuto R. Endoplasmic reticulum, Bcl-2 and Ca(2+) handling in apoptosis. Cell Calcium 32: 413-420, 2002.
97. Antonsson B. and Martinou J.C. The Bcl-2 protein family. Exp Cell Res 256:50-57, 2000.
98. Herr I. and Debatin K.M. Cellular stress response and apoptosis in cancer therapy. Blood 98: 2603-2614, 2001.
99. Hengartner M.O. The biochemistry of apoptosis. Nature 407: 770-776, 2000.
100.Wyllie A.H.,Moris R.G., Smith A.L. and Dunlop D. Chromatin cleavage in apoptosis：association with condensed chromatin morphology and dependence and dependence on macromolecular synthesis. Journal of pathology. 142：67-77, 1984.
101. Liu X., Kim C.N., Yang J., Jemmerson R. and Wang X. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 86: 147-157, 1996.
102. Yang J., Liu X., Bhalla K., Kim C.N., Ibrado A.M., Cai J., Peng T.I., Jones D.P. and Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 275: 1129-1132, 1997.
103. Kluck R.M., Bossy-Wetzel E., Green D.R. and Newmeyer D.D. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 275: 1132-1136, 1997.
104. Cai J., Yang J. and Jones D.P. Mitochondrial control of apoptosis: the role of cytochrome c. Biochim Biophys Acta 1366: 139-149, 1998.
105. Lindahl T., Satoh M. S., Poirier G. G. and Klungland A. Post –translational modification of poly(ADP-ribose) polymerase induced by DNA strand breaks. TIBS : 405-411, 1995.
106. Nicholson D. W. and Thornberry N. A. Apoptosis. Life and death decisions. Science: 299-306, 1997.
107.Song T.Y. and Yen G.C. Antioxidant properties of Antrodia camphorata in aubmerged culture. J Agr Food Chem 50, 3322-3327, 2002.
108.Dubois R.N., Abramon S.B., Crofford L., Gupta R.A., Simon L.S., Van De Putte L.B. and LipskyP.E. Cyclooxygenase in biology and disease. FASEB 12 : 1063-1073, 1998.
109. Williams C.S., Mann M. and DuBois R.N.The role of cyclooxygenases in inflammation, cancer and development.Oncogen 8 : 7908-7946, 1999.
110. Williams C.S., Tsujii M., Reese J., Sudhansu K. and DuBois R.N. Host cyclooxygenase-2 modulates carcinoma growth. J Clin Invest 105:1589-1594, 2000.
111. Stephen M. and Fitzpatrick F.A. Cyclooxygenase-2 and carcinogenesis . Biochimica et Biophysica Acta 1470 : M69-78, 2000.
112. Gupta R.A. and DuBois R.N. Colorectal cancer prevention and treatment by inhibition of cyclooxygenase-2. Nat Rev Cancer 1:11-21, 2001.
113. Howe L.R., Subbaramaiah K., Brown A.M.C. and Dannenberg A.J. Cyclooxygenase-2: a target for the prevention and treatment of breast cancer. Endoc Relat Cancer 8: 97-114, 2001.
114. Singh B. and Lucci A. Role of cyclooxygenase-2 in breast
cancer. J Surg Res 108: 173-179, 2002.
115. Pai R., Nakamura T., Moon W.S. and Tarnawski A.S. Prostaglandins promote colon cancer cell invasion; signaling by cross-talk between two distinct growth factor receptors. FASEB J 17:1640–7, 2003.
116. Chang S.H., Liu C.H. and Conway R., Role of prostaglandin E2-dependent angiogenic switch in cyclooxygenase 2-induced breast cancer progression. Proc Natl Acad Sci U S A 101:591–6, 2004.
117. Fujita H., Koshida K. and Keller E.T. Cyclooxygenase-2 promotes prostate cancer progression. Prostate 53:232–40, 2002.
118. Basu G.D., Pathangey L.B., Tinder T.L., Gendler S.J. and Mukherjee P. Mechanisms underlying the growth inhibitory effects of the cyclo-oxygenase-2 inhibitor celecoxib in human breast cancer cells. Breast Cancer Res. 7(4):R422-35, 2005.
119. Basu G.D., Pathangey L.B., Tinder T.L., Lagioia M., Gendler S.J. and Mukherjee P. Cyclooxygenase-2 inhibitor induces apoptosis in breast cancer cells in an in vivo model of spontaneous metastatic breast cancer.Mol Cancer Res. 2(11):632-42, 2004.
120. Meric J.B., Rottey S., Olaussen K., Soria J.C., Khayat D., Rixe O. and Spano J.P. Cyclooxygenase-2 as a target for anticancer drug development. Crit Rev Oncol Hematol. 2006
121.Tak P.P. and Firestein G.S. NF-κB：a key role in inflammatory diseases. J Clin Invest 107：7-11, 2001.
122.Schmid R.M. and Adler G. NF-κB/Rel/IκB：implications in gastrointestinal diseases. Gastroenterology 118：1208-1228, 2000.
123.Ghosh S., May M.J. and Kopp E.B. NF-κB and Rel proteins：evolutionarily conserved mediators of immune responses Annu Rev Immunol 16：225-260, 1998.
124.Greten F.R., Weber C.K., Greten T.F., Schneider G., Wagner M., Adler G. and Schmid R.M. Stat3 and NF-kappa B activation prevents apoptosis in pancreatic carcinogenesis. Gastroenterology 123：2052-2063, 2002.
125.Zang G. and Ghosh S. Toll-Like receptor-mediated NF-κB activation：a phylogenetically conversed paradigm in innate immunity. J Clin Invest 107：13-19, 2001.
126.May M.J. and Ghosh S. Rel/NF-κB and IκB proteins：anoverview. Semin Cancer Biol 8：63-731, 1997
127.Wang Z., Castresana M.R., Detmer K. and Newman W.H. An IκB-α mutant inhibits cytokine gene expression and proliferation in human vascular cmoothmuscle cells. J Surg Res. 102：198-206, 2002.
128.Karin M. and Ben-Neriah Y. Phosphorylation meets ubiquitination：the control of NF-κB activity. Annu Rev Immunol.18：621-663, 2000.
129.May M.J. and Ghosh S. Signal transduction through NF-κB. Immunol today 19：80-88, 1998.
130.Flohe L., Brigelius-Flohe R., Saliou C., Traber M.G. and Packer L. Redox regulation of NF-κB activation. Free Radic Biol Med. 22：1115-1126, 1997.
131.Rothwarf D.M., Zandi E., Natoli G. and Karin M. IKK-γ is an essential regulatory subunit of the IκB kinase complex. Nature 395：297-300, 1998.
132.Nakano H., Shindo M., Sakon S., Nishinaka S., Mihara M., Yagita H. and Okumura K. Differential regulation of IκB kinases alpha and beta by two upatream kinases, NF-κB-inducing kinases and mitogen-activated protein kinases/ERK kinase kinase-1. Proc Natl Acand Sci USA 95：3537-3542, 1998.
133.Karin M. and Lin A. NF-κB at the crossroads of life and death. Nat. Immunol. 3：221-227, 2002.
134.Pham C. G., C. Bubici F. Zazzeroni S. Papa J. Jones K. Alvarez S. Jayawardena E. De Smaele R. Cong C. Beaumont F. M. Torti S.V. and Franzoso G. ferritin heavy chain upregulation by NF-κB inhibits TNF-α-induced apoptosis by suppressing reactive oxygen species. Cell 119：529-542, 2004.
135.Shakhov A. N., Collart M. A., Vassalli P., Nedospasov S. A. and Jongeneel C.V. Kappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factoralpha genen in primary macrophages. J Exp. Med. 171：35-47, 1990.
136.Fujioka S., Schmidt C., Sclabas G. M., Li Z., Pelicano H., Peng B., Yao A., Niu J., Zang W., Evans D. B., Abbruzzese J. L., Huang P., and Chiao P. J. Stabilization of p53 is a novel mechanism for proapoptotic function of NF-κB. J. Biol. Chem. 279：27549-27559, 2004.
137.Luo, J.L., Kamata H. and Karin M. IKK/NF-kappaB signaling：balancing life and death – a new approach to cancer therapy. J. Clin. Invest. 115：2625-2632, 2005.
138. Sheng H., Shao J. and Morrow J.D. Modulation of apoptosis and Bcl-2 expression by prostaglandin E2 in human colon cancer cells. Cancer Res 58:362–6, 1998.
139.Lamkanfi M., Declercq W., Vanden Berghe T. and Vandenabeele P. Caspases leave the beaten track: caspase-mediated activation of NF-kappaB. J Cell Biol. Apr 24;173(2):165-71, 2006.
140. Brozovic S., Sahoo R., Barve S., Shiba H., Uriarte S., Blumberg R.S and Kinane D.F. Porphyromonas gingivalis enhances FasL expression via up-regulation of NFkappaB-mediated gene transcription and induces apoptotic cell death in human gingival epithelial cells. Microbiology. 152(Pt 3):797-806, 2006.
141.Kashfi K. and Rigas B. Is COX-2 a ‘collateral’ target in cancer prevention? Biochem. Soc. Trans. 33:724–727, 2005.
142.Suzuki T. and Tsukamoto I. Arsenite induces apoptosis in hepatocytes through an enhancement of the activation of Jun N-terminal kinase and p38 mitogen-activated protein kinase caused by partial hepatectomy.Toxicol Lett. 2006
143. Johnson G.L. and Lapadat R. Mitogen-activated protein kinase pathwaysmediated by ERK, JNK and p38 protein kinases. Science (Wash DC) 298:1911–1912, 2002
144. Park M.T., Choi J.A., Kim M.J., Um H.D., Bae S., Kang C.M., Cho C.K., Kang S., Chung H.Y., Lee Y.S., and Lee S.J. Suppression of extracellular signal-related kinase and activation of p38 MAPK are two critical events leading to caspase-8- and mitochondria-mediated cell death in phytosphingosine - treated human cancer cells.J Biol Chem 278:50624–50634, 2003.
145. Olson J.M. and Hallahan A.R. p38 MAP kinase: a convergence point in cancer therapy. Trends Mol Med 10:125–129, 2004.
146. Bacus S.S., Gudkov A.V., Lowe M., Lyass L., Yung Y., Komarov A.P., Keyomarsi K., Yarden Y. and Seger R. Taxol-induced apoptosis depends on MAP kinase pathways (ERK and p38) and is independent of p53. Oncogene 20:147–155, 2001.
147. She Q.B., Bode A.M., Ma W.Y., Chen N.Y. and Dong Z. Resveratrol-induced activation of p53 and apoptosis is mediated by extracellular-signal-regulated protein kinases and p38 kinase. Cancer Res 61:1604–1610, 2001
148. Nguyen T.T., Tran E., Nguyen T.H., Do P.T., Huynh T.H., and Huynh H. The role of activated MEK-ERK pathway in quercetin-induced growth inhibition and apoptosis in A549 lung cancer cells. Carcinogenesis 25:647–659, 2004.
149. Yu W., Liao Q.Y., Hantash F.M., Sanders B.G. and Kline K. Activation of extracellular signal-regulated kinase and c-Jun-NH(2)-terminal kinase but not p38 mitogen-activated protein kinases is required for RRR-alpha-tocopheryl succinateinduced apoptosis of human breast cancer cells. Cancer Res 61:6569–6576, 2001.
150.Lohr D., Tatchell K. and Van Holde K.E.On the occurrence of nucleosome phasing in chromatin. Cell. 12(3):829-36, 1977.
151.Detrick-Hooks B., Borsos T. and Rapp H.J. Quantitative comparison of techniques used to measure complement-mediated cytotoxicity of nucleated cells. J Immunol. 114:287-90, 1975.
152.Ostling O., Johanson K.J., Blomquist E. and Hagelqvist E. DNA damage in clinical radiation therapy studied by microelectrophoresis in single tumour cells. A preliminary report. Acta Oncol. 26(1):45-8, 1987.
153. Burnette W.N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 112(2):195-203, 1981.
154.Benotmane A.M., Hoylaerts M.F., Collen D. and Belayew A. Nonisotopic quantitative analysis of protein-DNA interactions at equilibrium.Anal Biochem. 250(2):181-5, 1997.