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研究生:陳玲儀
研究生(外文):Ling-Yi Chen
論文名稱:前處理牛樟芝之乙醇萃取物能加強Amphotericin B細胞毒性之反應研究
論文名稱(外文):Pretreatment with an ethanolic extract ofTaiwanofungus camphoratus (Antrodia camphorata)enhances the cytotoxicity of Amphotericin B
指導教授:蘇慶華蘇慶華引用關係
指導教授(外文):Ching-Hua Su
學位類別:博士
校院名稱:臺北醫學大學
系所名稱:醫學人文研究所
學門:人文學門
學類:其他人文學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:79
中文關鍵詞:樟芝細胞凋亡兩性黴素細胞週期抗黴菌藥
外文關鍵詞:Taiwanofungus camphoratusapoptosisamphotericin Bcell cycleantifungal agents
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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

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