臺灣博碩士論文加值系統

CONTENTS
Acknowledgements................................................................................... II
Abbreviations............................................................................................ III
Contents..................................................................................................... V
Contents of Figures ................................................................................. VII
Abstract (Chinese).................................................................................... IX
Abstract (English).................................................................................... X
1. Introduction.......................................................................................... 1
1.1 Taiwanofungus camphoratus (Antrodia camphorata) ............ 1
1.2 Amphotericin B........................................................................ 5
1.3 Programmed cell death............................................................ 7
1.4 Cell cycle regulation................................................................ 9
1.5 Complementary medicine......................................................... 10
1.6 Melanoma................................................................................. 11
1.7 Osteosarcoma........................................................................... 12
1.8 Colon cancer............................................................................ 13
1.9 Specific aims............................................................................ 14
2. Materials and Methods........................................................................ 16
2.1 AmB and TCEE preparation.................................................... 16
2.2 Cell lines and culture............................................................... 16
2.3 MTT assay................................................................................ 17
2.4 Western blot analysis............................................................... 18
2.5 Annexin V-PI staining.............................................................. 19
2.6 Cell cycle analysis.................................................................... 20
2.7 Mitochondrial membrane potential analysis........................... 21
2.8 HT29-derived xenografts animal model.................................. 22
2.9 Statistical analysis................................................................... 23
3. Results................................................................................................... 24
3.1 Cytotoxicity of AmB and TCEE................................................ 25
3.2 Induction of apoptosis by combined treatment with AmB and
TCEE........................................................................................26
3.3 Cell cycle arrest induced by combined treatment with AmB
and TCEE..............................................................................27
3.4 Inhibitory effect of TCEE and AmB on xenograft growth in
nude mice...............................................................................28
4. Discussion.............................................................................................. 30
5. Conclusion and Perspective................................................................. 37
6. References............................................................................................. 39
7. Figures................................................................................................... 50
8. Appendix (List of publications)........................................................... 69
Contents of Figures
Figure 1. The cytotoxic effect of AmB.................................................. 50
Figure 2. The cytotoxic effect of TCEE................................................. 51
Figure 3. The cytotoxic effects of AmB and TCEE used in
combination............................................................................52
Figure 4. AmB does not induce apoptosis in the HT29 and
RPMI7951 cells.................................................................53
Figure 5. AmB does not cause cell death in the HT29 and RPMI7951
cells....................................................................................54
Figure 6. The apoptotic effects of TCEE and AmB............................... 55
Figure 7. Combined treatment of AmB and TCEE results in cell
death....................................................................................56
Figure 8. Combined treatment of AmB and TCEE induces
apoptosis..............................................................................57
Figure 9. Loss of the mitochondrial membrane potential in cells
treated with a combination of TCEE and AmB....................58
Figure 10. Combined treatment of TCEE and AmB induces loss of the
mitochondrial membrane potential in a dose-dependent
manner.................................................................................59
Figure 11. The expression of anti-apoptotic and pro-apoptotic proteins
in cells treated with a combination of TCEE and AmB..........60
Figure 12. Increased levels of anti-apoptotic proteins and decreased
levels of pro-apoptotic proteins in cells treated with a
combination of TCEE and AmB............................................. 61
Figure 13. The Bax-to-Bcl-2 ratio increases in cells treated with a
combination of TCEE and AmB...........................................62
Figure 14. The combined treatment of TCEE and AmB alters cell cycle
distribution............................................................................63
Figure 15. Induction of G2/M arrest in HT29 cells treated with TCEE
and AmB.............................................................................64
Figure 16. The expression of cell-cycle associated proteins in cells
treated with a combination of TCEE and AmB.....................65
Figure 17. Alteration of cell cycle associated protein under TCEE and
AmB treatment......................................................................66
Figure 18. The therapeutic effects of TCEE and AmB in nude mice
bearing human HT29 xenografts..........................................67
Figure 19. Inhibition of HT29 xenografts by a combination of TCEE
and AmB...............................................................................68

1. Chang, T. T., Antrodia cinnamomea sp. Nov. on Cinnamomum
kanehirai in Taiwan. Mycol Res 1995, 99, 756-758.
2. Wu, S. H., Antrodia camphorata (“niu-chang-chih”), new
combination of a medicinal fungus in Taiwan. Bot Bull Acad Sin 1997, 38, 273-275.
3. Chang, T. T.; Chou, W. N., Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. Mycol Res 1995, 99, (6),
756-758.
4. Chen, J. C.; Lin, W. H.; Chen, C. N.; Sheu, S. J.; Huang, S. J.; Chen, Y. L., Development of Antrodia camphorata mycelium with submerged culture. Fungal Sciences 2001, 16, (7-22).
5. Shen, C. C., New ergostane and lanostane from Antrodia
camphorata. J Chin Med 2003, 14, 247-258.
6. Chen, C. J.; Su, C. H.; Lan, M. H., Study on solid cultivation and bioactivity of Antrodia camphorata. Fungal Sciences 2001, 16, 65-72.
7. Zang, M.; Su, Q. H., Ganoderma comphoratum, a new taxon in genus Ganoderma from Taiwan, China,. Acta Botanica Yunnanica 1990, 12, 395-396.
8. Chen, J. J.; Lin, W. J.; Liao, C. H.; Shieh, P. C., Anti-inflammatory benzenoids from Antrodia camphorata. J Nat Prod 2007, 70, (6),
989-92.
9. Hsiao, G.; Shen, M. Y.; Lin, K. H.; Lan, M. H.; Wu, L. Y.; Chou, D. S.; Lin, C. H.; Su, C. H.; Sheu, J. R., Antioxidative and hepatoprotective effects of Antrodia camphorata extract. J Agric Food Chem 2003, 51, (11), 3302-8.
10. Rao, Y. K.; Fang, S. H.; Tzeng, Y. M., Evaluation of the
anti-inflammatory and anti-proliferation tumoral cells activities of Antrodia camphorata, Cordyceps sinensis, and Cinnamomum osmophloeum bark extracts. J Ethnopharmacol 2007, 114, (1), 78-85.
11. Liu, J. J.; Huang, T. S.; Hsu, M. L.; Chen, C. C.; Lin, W. S.; Lu, F. J.; Chang, W. H., Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. Toxicol Appl Pharmacol 2004, 201, (2), 186-93.
12. Geethangili, M.; Tzeng, Y. M., Review of Pharmacological Effects of Antrodia camphorata and its Bioactive Compounds. Evid Based Complement Alternat Med 2009.
13. Bishayee, A.; Ahmed, S.; Brankov, N.; Perloff, M., Triterpenoids as potential agents for the chemoprevention and therapy of breast cancer. Front Biosci 2011, 16, 980-96.
14. Yeh, C. T.; Rao, Y. K.; Yao, C. J.; Yeh, C. F.; Li, C. H.; Chuang, S. E.; Luong, J. H.; Lai, G. M.; Tzeng, Y. M., Cytotoxic triterpenes from Antrodia camphorata and their mode of action in HT-29 human colon cancer cells. Cancer Lett 2009, 285, (1), 73-9.
15. Zuco, V.; Supino, R.; Righetti, S. C.; Cleris, L.; Marchesi, E.; Gambacorti-Passerini, C.; Formelli, F., Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells. Cancer Lett 2002, 175, (1), 17-25.
16. Chen, Y.-C.; Ho, H.-O.; Su, C.-H.; Sheu, M.-T., Anticancer Effects of Taiwanofungusnext termprevious termcamphoratusnext term Extracts, Isolated Compounds and its Combinational use. 2010, 2, (6), 274-81.
17. Ho, C. M.; Huang, C. C.; Huang, C. J.; Cheng, J. S.; Chen, I. S.; Tsai, J. Y.; Jiann, B. P.; Tseng, P. L.; Kuo, S. J.; Jan, C. R., Effects of antrodia camphorata on viability, apoptosis, and [Ca2+]i in PC3 human prostate cancer cells. Chin J Physiol 2008, 51, (2), 78-84.
18. Yang, H. L.; Chen, C. S.; Chang, W. H.; Lu, F. J.; Lai, Y. C.; Chen, C. C.; Hseu, T. H.; Kuo, C. T.; Hseu, Y. C., Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by Antrodia camphorata. Cancer Lett 2006, 231, (2), 215-27.
19. Lu, Z. M.; Tao, W. Y.; Zou, X. L.; Fu, H. Z.; Ao, Z. H., Protective effects of mycelia of Antrodia camphorata and Armillariella tabescens in submerged culture against ethanol-induced hepatic toxicity in rats. J Ethnopharmacol 2007, 110, (1), 160-4.
20. Liu, D. Z.; Liang, Y. C.; Lin, S. Y.; Lin, Y. S.; Wu, W. C.; Hou, W. C.; Su, C. H., Antihypertensive activities of a solid-state culture of Taiwanofungus camphoratus (Chang-chih) in spontaneously hypertensive rats. Biosci Biotechnol Biochem 2007, 71, (1), 23-30.
21. Song, T. Y.; Yen, G. C., Antioxidant properties of Antrodia camphorata in submerged culture. J Agric Food Chem 2002, 50, (11), 3322-7.
22. Chen, C. C.; Shiao, Y. J.; Lin, R. D.; Shao, Y. Y.; Lai, M. N.; Lin, C. C.; Ng, L. T.; Kuo, Y. H., Neuroprotective diterpenes from the fruiting body of Antrodia camphorata. J Nat Prod 2006, 69, (4), 689-91.
23. Aydemir, G., Research on Nutrition and Cancer: The Importance of the Standardized Dietary Assessments. Asian Pac J Cancer Prev 2002, 3, (2), 177-180.
24. Gallis, H. A.; Drew, R. H.; Pickard, W. W., Amphotericin B: 30 years of clinical experience. Rev Infect Dis 1990, 12, (2), 308-29.
25. Bodey, G. P.; Anaissie, E. J.; Elting, L. S.; Estey, E.; O''Brien, S.; Kantarjian, H., Antifungal prophylaxis during remission induction therapy for acute leukemia fluconazole versus intravenous amphotericin B. Cancer 1994, 73, (8), 2099-106.
26. Pienaar, E. D.; Young, T.; Holmes, H., Interventions for the prevention and management of oropharyngeal candidiasis associated with HIV infection in adults and children. Cochrane Database Syst Rev 2010, (11), CD003940.
27. Tollemar, J.; Hockerstedt, K.; Ericzon, B. G.; Jalanko, H.; Ringden, O., Liposomal amphotericin B prevents invasive fungal infections in liver transplant recipients. A randomized, placebo-controlled study. Transplantation 1995, 59, (1), 45-50.
28. Dentinger, P. J.; Swenson, C. F.; Anaizi, N. H., Stability of amphotericin B in an extemporaneously compounded oral suspension. Am J Health Syst Pharm 2001, 58, (11), 1021-4.
29. Bortner, C. D.; Hughes, F. M., Jr.; Cidlowski, J. A., A primary role for K+ and Na+ efflux in the activation of apoptosis. J Biol Chem 1997, 272, (51), 32436-42.
30. Cohen, B. E., Amphotericin B membrane action: role for two types of ion channels in eliciting cell survival and lethal effects. J Membr Biol 2010, 238, (1-3), 1-20.
31. Beketic-Oreskovic, L.; Osmak, M., Modulation of resistance to cisplatin by amphotericin B and aphidicolin in human larynx carcinoma cells. Cancer Chemother Pharmacol 1995, 35, (4), 327-33.
32. Kikkawa, F.; Kojima, M.; Oguchi, H.; Maeda, O.; Ishikawa, H.; Tamakoshi, K.; Mizuno, K.; Kawai, M.; Suganuma, N.; Tomoda, Y., Potentiating effect of amphotericin B on five platinum anticancer drugs in human cis-diamminedichloroplatinum (II) sensitive and resistant ovarian carcinoma cells. Anticancer Res 1993, 13, (4), 891-6.
33. Feigin, A. M., Selective modification of sterol composition of hepatomas: new opportunities for chemotherapy. Med Hypotheses 1999, 52, (5), 383-8.
34. Meier, P.; Finch, A.; Evan, G., Apoptosis in development. Nature 2000, 407, (6805), 796-801.
35. Wyllie, A. H.; Kerr, J. F.; Currie, A. R., Cell death: the significance of apoptosis. Int Rev Cytol 1980, 68, 251-306.
36. Wyllie, A. H., Apoptosis: an overview. Br Med Bull 1997, 53, (3), 451-65.
37. Nicholson, D. W., From bench to clinic with apoptosis-based therapeutic agents. Nature 2000, 407, (6805), 810-6.
38. Chiang, C. T.; Way, T. D.; Lin, J. K., Sensitizing
HER2-overexpressing cancer cells to luteolin-induced apoptosis through suppressing p21(WAF1/CIP1) expression with rapamycin. Mol Cancer Ther 2007, 6, (7), 2127-38.
39. Bold, R. J.; Termuhlen, P. M.; McConkey, D. J., Apoptosis, cancer and cancer therapy. Surg Oncol 1997, 6, (3), 133-42.
40. Ashkenazi, A.; Dixit, V. M., Apoptosis control by death and decoy receptors. Curr Opin Cell Biol 1999, 11, (2), 255-60.
41. Green, D. R.; Reed, J. C., Mitochondria and apoptosis. Science 1998, 281, (5381), 1309-12.
42. Sheridan, J. P.; Marsters, S. A.; Pitti, R. M.; Gurney, A.; Skubatch, M.; Baldwin, D.; Ramakrishnan, L.; Gray, C. L.; Baker, K.; Wood, W. I.; Goddard, A. D.; Godowski, P.; Ashkenazi, A., Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 1997, 277, (5327), 818-21.
43. Kuwana, T.; Newmeyer, D. D., Bcl-2-family proteins and the role of mitochondria in apoptosis. Curr Opin Cell Biol 2003, 15, (6), 691-9.
44. Nicholson, D. W.; Ali, A.; Thornberry, N. A.; Vaillancourt, J. P.; Ding, C. K.; Gallant, M.; Gareau, Y.; Griffin, P. R.; Labelle, M.; Lazebnik, Y. A.; et al., Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 1995, 376, (6535), 37-43.
45. Li, P.; Nijhawan, D.; Budihardjo, I.; Srinivasula, S. M.; Ahmad, M.; Alnemri, E. S.; Wang, X., Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997, 91, (4), 479-89.
46. Vermeulen, K.; Van Bockstaele, D. R.; Berneman, Z. N., The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif 2003, 36, (3), 131-49.
47. Maddika, S.; Ande, S. R.; Panigrahi, S.; Paranjothy, T.; Weglarczyk, K.; Zuse, A.; Eshraghi, M.; Manda, K. D.; Wiechec, E.; Los, M., Cell survival, cell death and cell cycle pathways are interconnected: implications for cancer therapy. Drug Resist Updat 2007, 10, (1-2), 13-29.
48. Gray-Schopfer, V. C.; da Rocha Dias, S.; Marais, R., The role of B-RAF in melanoma. Cancer Metastasis Rev 2005, 24, (1), 165-83.
49. Balch, C. M.; Buzaid, A. C.; Soong, S. J.; Atkins, M. B.; Cascinelli, N.; Coit, D. G.; Fleming, I. D.; Gershenwald, J. E.; Houghton, A., Jr.; Kirkwood, J. M.; McMasters, K. M.; Mihm, M. F.; Morton, D. L.; Reintgen, D. S.; Ross, M. I.; Sober, A.; Thompson, J. A.; Thompson, J. F., Final version of the American Joint Committee on
Cancer staging system for cutaneous melanoma. J Clin Oncol 2001, 19, (16), 3635-48.
50. Soengas, M. S.; Lowe, S. W., Apoptosis and melanoma
chemoresistance. Oncogene 2003, 22, (20), 3138-51.
51. Meyers, P. A.; Gorlick, R., Osteosarcoma. Pediatr Clin North Am 1997, 44, (4), 973-89.
52. Parkin, D. M.; Bray, F.; Ferlay, J.; Pisani, P., Global cancer statistics, 2002. CA Cancer J Clin 2005, 55, (2), 74-108.
53. Allen, W. L.; Coyle, V. M.; Johnston, P. G., Predicting the outcome of chemotherapy for colorectal cancer. Curr Opin Pharmacol 2006, 6, (4), 332-6.
54. Almyroudis, N. G.; Segal, B. H., Antifungal prophylaxis and therapy in patients with hematological malignancies and
hematopoietic stem cell transplant recipients. Expert Rev Anti Infect Ther 2010, 8, (12), 1451-66.
55. Morikage, T.; Bungo, M.; Inomata, M.; Yoshida, M.; Ohmori, T.; Fujiwara, Y.; Nishio, K.; Saijo, N., Reversal of cisplatin resistance with amphotericin B in a non-small cell lung cancer cell line. Jpn J Cancer Res 1991, 82, (6), 747-51.
56. Oki, E.; Sakaguchi, Y.; Toh, Y.; Oda, S.; Maehara, Y.; Yamamoto, N.; Sugimachi, K., Induction of apoptosis in human tumour xenografts after oral administration of uracil and tegafur to nude mice bearing tumours. Br J Cancer 1998, 78, (5), 625-30.
57. Hsu, Y. L.; Kuo, Y. C.; Kuo, P. L.; Ng, L. T.; Kuo, Y. H.; Lin, C. C., Apoptotic effects of extract from Antrodia camphorata fruiting bodies in human hepatocellular carcinoma cell lines. Cancer Lett 2005, 221, (1), 77-89.
58. Fujii, G.; Chang, J. E.; Coley, T.; Steere, B., The formation of amphotericin B ion channels in lipid bilayers. Biochemistry 1997, 36, (16), 4959-68.
59. Huang, W.; Zhang, Z.; Han, X.; Tang, J.; Wang, J.; Dong, S.; Wang, E., Ion channel behavior of amphotericin B in sterol-free and cholesterol- or ergosterol-containing supported phosphatidylcholine bilayer model membranes investigated by electrochemistry and spectroscopy. Biophys J 2002, 83, (6), 3245-55.
60. Feigin, A. M.; Gnanik, T.; Pasechnik, V. I.; Flerov, M. N.; Bogoslovskii, N. A., [Role of sterols in the interaction of polyene antibiotics with lipid membranes]. Antibiotiki 1981, 26, (7), 522-6.
61. Feigin, A. M.; Flerov, M. N.; Gianik, T.; Pasechnik, V. I.; Belousova, II, [Change of current-voltage characteristics of amphotericin B ion channels accompanying its incorporation into the membrane]. Biofizika 1982, 27, (2), 331-3.
62. Feigin, A. M.; Teeter, J. H.; Brand, J. G., The influence of sterols on the sensitivity of lipid bilayers to melittin. Biochem Biophys Res Commun 1995, 211, (1), 312-7.
63. Charbonneau, C.; Fournier, I.; Dufresne, S.; Barwicz, J.; Tancrede, P., The interactions of amphotericin B with various sterols in relation to its possible use in anticancer therapy. Biophys Chem 2001, 91, (2), 125-33.
64. Yano, T.; Itoh, Y.; Kawamura, E.; Maeda, A.; Egashira, N.; Nishida, M.; Kurose, H.; Oishi, R., Amphotericin B-induced renal tubular cell injury is mediated by Na+ Influx through ion-permeable pores and subsequent activation of mitogen-activated protein kinases and elevation of intracellular Ca2+ concentration. Antimicrob Agents
Chemother 2009, 53, (4), 1420-6.
65. Varlam, D. E.; Siddiq, M. M.; Parton, L. A.; Russmann, H., Apoptosis contributes to amphotericin B-induced nephrotoxicity. Antimicrob Agents Chemother 2001, 45, (3), 679-85.
66. Marklund, L.; Henriksson, R.; Grankvist, K., Cisplatin-induced apoptosis of mesothelioma cells is affected by potassium ion flux modulator amphotericin B and bumetanide. Int J Cancer 2001, 93, (4), 577-83.
67. Wu, H.; Pan, C. L.; Yao, Y. C.; Chang, S. S.; Li, S. L.; Wu, T. F., Proteomic analysis of the effect of Antrodia camphorata extract on human lung cancer A549 cell. Proteomics 2006, 6, (3), 826-35.
68. Li, S. L.; Huang, Z. N.; Hsieh, H. H.; Yu, W. C.; Tzeng, W. Y.; Lee, G. Y.; Chen, Y. P.; Chang, C. Y.; Chuu, J. J., The augmented anti-tumor effects of Antrodia camphorata co-fermented with Chinese medicinal herb in human hepatoma cells. Am J Chin Med 2009, 37, (4), 771-83.
69. Chang, C. Y.; Huang, Z. N.; Yu, H. H.; Chang, L. H.; Li, S. L.; Chen, Y. P.; Lee, K. Y.; Chuu, J. J., The adjuvant effects of Antrodia Camphorata extracts combined with anti-tumor agents on multidrug resistant human hepatoma cells. J Ethnopharmacol 2008, 118, (3), 387-95.
70. Chiu, A.; Shi, X. L.; Lee, W. K.; Hill, R.; Wakeman, T. P.; Katz, A.; Xu, B.; Dalal, N. S.; Robertson, J. D.; Chen, C.; Chiu, N.; Donehower, L., Review of chromium (VI) apoptosis,cell-cycle-arrest, and carcinogenesis. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 2010, 28, (3), 188-230.
71. Herman-Antosiewicz, A.; Singh, S. V., Signal transduction pathways leading to cell cycle arrest and apoptosis induction in cancer cells by Allium vegetable-derived organosulfur compounds: a review. Mutat Res 2004, 555, (1-2), 121-31.
72. Kastan, M. B.; Bartek, J., Cell-cycle checkpoints and cancer. Nature 2004, 432, (7015), 316-23.
73. Coqueret, O., New roles for p21 and p27 cell-cycle inhibitors: a function for each cell compartment? Trends Cell Biol 2003, 13, (2), 65-70.
74. Li, Y. H.; Chung, H. C.; Liu, S. L.; Chao, T. H.; Chen, J. C., A novel inhibitory effect of Antrodia camphorata extract on vascular smooth muscle cell migration and neointima formation in mice. Int Heart J 2009, 50, (2), 207-20.
75. Ferguson, P. J.; Currie, C.; Vincent, M. D., Enhancement of platinum-drug cytotoxicity in a human head and neck squamous cell carcinoma line and its platinum-resistant variant by liposomal amphotericin B and phospholipase A2-II. Drug Metab Dispos 1999, 27, (12), 1399-405.