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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40830完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林淑萍 | |
| dc.contributor.author | I-Lin Lee | en |
| dc.contributor.author | 李依霖 | zh_TW |
| dc.date.accessioned | 2021-06-14T17:02:23Z | - |
| dc.date.available | 2010-08-08 | |
| dc.date.copyright | 2008-08-08 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-29 | |
| dc.identifier.citation | Ashkenazi, A. (2008) Targeting the extrinsic apoptosis pathway in cancer. Cytokine Growth Factor Rev. 19, 325-331
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40830 | - |
| dc.description.abstract | 本研究主要探討靈芝三萜類Ganoderic acid T (GAT)的抗癌活性，使用GAT針對不同的癌細胞進行研究，觀察是否能促使細胞凋亡或細胞老化。發現GAT可抑制人類血癌細胞株HL-60、肝癌細胞株Hep-G2和Huh7細胞的生長，其IC50分別為35.2，20.3及75.7，而對人類正常週邊血液單核球細胞沒有影響。在HL-60細胞的研究中，藉由劉氏染色法和TUNEL實驗、粒線體膜電位分析、細胞週期分析、7-AAD染色及流式細胞分析，caspase-8、caspase-9及caspase-3活性分析這一系列和細胞凋亡有關的實驗，在Liu’s stain中發現有apoptotic bodies，在TUNEL實驗中有發現DNA斷裂，在粒線體膜電位分析中發現粒線體膜電位的下降，在7-AAD實驗看到細胞膜的完整性降低導致7-AAD螢光染色程度升高，在caspase-9和caspase-3實驗看到caspases活性的上升，顯示GAT有誘導HL-60細胞凋亡的能力。在Huh7 細胞發現GAT會使細胞的粒腺體膜電位下降、細胞週期停滯在G2期。在HepG2的研究，初步觀察到GAT會使HepG2細胞產生老化的細胞型態。除此之外，將加過GAT的細胞培養液的回收，萃取其中的物質，藉由高效液相層析儀，再配合四級柱質譜儀分析後，發現仍可找到結構沒改變的GAT，可見GAT在細胞培養液中的穩定性。由以上結果顯示GAT有發展為抗癌藥物的可能性。 | zh_TW |
| dc.description.abstract | Ganoderic acid T (GAT), is a lanostanoid triterpene isolated from Ganoderma lucidum (Leyss.exFr.) Karst., and the aim of this study was to study the effects of GAT on some cancer cell lines. GAT was found to inhibit the growths of human leukemia cell line, HL-60 and hepatoma cell lines, HepG2 and Huh7, and the IC50 are 35.2, 20.3 and 75.7 μM respectively, while GAT showed no effect on non-proliferating human peripheral blood mononuclear cells. In HL-60 cells, Liu’s stain, TUNEL assay, MMP, cell cycle, 7-AAD, caspase-8, caspase-9 were performed. Apoptotic bodies were shown by Liu’s stain, TUNEL assay positive cells were observed in TUNEL assay, and 7-AAD positive cells were observed. Mitochrondrial membrane potential was decreased, and caspase-9 and 3 activities were increased in GAT treatment. In conclusion, GAT was found to induce HL-60 cell apoptosis. In Huh 7 cells, GAT was found to decrease mitochondrial membrane potential and induce cell cycle arrest in G2 phase. In HepG2 cells, preliminary data showed that GAT seemed to induce premature senescence of HepG2 cell. GAT was recovered from cultured medium, and HPLC and Q-TOF mass spectrometry was used to analyze GAT and the recovered fraction. The result showed that GAT is stable in cell culture. In the future, GAT might be recovered from culture medium. The results above indicate that GAT could be a potential anticancer drug. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-14T17:02:23Z (GMT). No. of bitstreams: 1 ntu-97-R95424011-1.pdf: 1937161 bytes, checksum: dee8c1f27315e4858c2671a552369a91 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 總 目 錄
中文摘要 5 英文摘要 6 縮寫表 7 一、序論 8 1.1 靈芝和靈芝酸的成分 8 1.2 細胞凋亡 9 1.3 細胞週期 10 1.4 細胞老化 11 1.5 研究目的 11 二、材料與方法 13 2.1 使用細胞株 13 2.2 GAT的來源、保存、定性及定量 13 2.3 細胞培養 13 2.4 細胞活性測定 14 2.5 人類週邊血單核球細胞 (PBMC) 的分離 14 2.6 劉式細胞染色 (Liu’s stain) 14 2.7 細胞粒線體膜電位的測定 15 2.8 細胞染色7-AAD的測定 15 2.9 TUNEL實驗 15 2.10 細胞內Caspases活性的測定 16 2.11 細胞週期分析 16 2.12 細胞內RNA的萃取 16 2.13 半定量反轉錄聚合酵素連鎖反應 17 2.14 衰老細胞的β-半乳糖苷酶染色 17 2.15 GAT在細胞培養液中的穩定性探討 18 三、結果 19 3.1 GAT的吸光圖譜 19 3.2 GAT對HL-60細胞具有抑制生長的效果，對PBMC則沒有影響。 19 3.3 GAT對HL-60細胞的影響。 19 3.3a GAT處理可觀察到HL-60細胞產生類似凋亡小體的細胞核濃染。 19 3.3b GAT處理可使HL-60細胞生長週期停滯於S期，並促進HL-60細胞的subG1百分比上升。 19 3.3c GAT處理可引發HL-60細胞DNA斷裂。 20 3.3d GAT處理可引發HL-60細胞膜通透性上升。 20 3.3e GAT處理可引發HL-60細胞粒線體膜電位的下降。 20 3.3f GAT處理可引發caspase-9的活化，不活化caspase-8。 21 3.3g GAT處理可引發caspase-3的活化。 21 3.3h GAT無法引發Bcl-2量的下降及E2F量的上升。 21 3.4 GAT對HepG2和Huh7細胞具有抑制生長的效果。 22 3.5 GAT會使Huh7細胞粒線體膜電位下降及細胞週期停滯在G2期。 22 3.6 GAT可能會引發HepG2細胞老化。 22 3.7 GAT在細胞培養液中的穩定性的探討。 23 四、討論 24 五、未來發展 27 六、參考文獻 28 藥品清單 48 圖 表 目 錄 圖1. (A) Ganoderic acid T (GAT)的化學結構式。 32 圖2. GAT的吸光光譜。 33 圖3. GAT對HL-60細胞的生長抑制作用。 34 圖4. GAT使HL-60細胞產生類似凋亡小體。 35 圖5. GAT使HL-60細胞週期停滯在S期，並促進HL-60細胞的subG1百分比上升。 36 圖6. GAT引發HL-60細胞DNA斷裂。 37 圖7. GAT增加HL-60細胞的細胞膜通透性。 38 圖8. GAT引發HL-60細胞粒線體膜電位的下降。 39 圖9. GAT引發HL-60細胞caspase-9的活化，而非caspase-8的活化。 40 圖10. GAT引發HL-60細胞caspase-3的活化。 41 圖11. GAT對HL-60細胞凋亡和生長相關分子基因表現的影響。 42 圖12. GAT對Huh7細胞的生長抑制作用。 43 圖13. GAT使Huh7細胞粒線體膜電位下降。 44 圖14. GAT對HepG2細胞的生長抑制作用。 45 圖15. GAT引發HepG2細胞的快速老化。 46 圖16. GAT在細胞培養液中具有穩定性。 47 | |
| dc.language.iso | zh-TW | |
| dc.subject | Huh7 | zh_TW |
| dc.subject | HepG2 | zh_TW |
| dc.subject | GAT | zh_TW |
| dc.subject | 細胞凋亡 | zh_TW |
| dc.subject | 細胞週期停滯 | zh_TW |
| dc.subject | 細胞老化 | zh_TW |
| dc.subject | 靈芝酸 | zh_TW |
| dc.subject | HL-60 | zh_TW |
| dc.subject | 赤芝 | zh_TW |
| dc.subject | Ganoderma | en |
| dc.subject | Huh7 | en |
| dc.subject | HepG2 | en |
| dc.subject | HL-60 | en |
| dc.subject | senescence | en |
| dc.subject | cell cycle arrest | en |
| dc.subject | apoptosis | en |
| dc.subject | GAT | en |
| dc.subject | Ganoderic acid | en |
| dc.title | 靈芝酸GAT的抗癌活性研究 | zh_TW |
| dc.title | Studies on anticancer activity of ganoderic acid T | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 歐樂君,胡忠怡,邱義源 | |
| dc.subject.keyword | 赤芝,靈芝酸,GAT,細胞凋亡,細胞週期停滯,細胞老化,HL-60,HepG2,Huh7, | zh_TW |
| dc.subject.keyword | Ganoderma,Ganoderic acid,GAT,apoptosis,cell cycle arrest,senescence,HL-60,HepG2,Huh7, | en |
| dc.relation.page | 48 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-07-29 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
| 顯示於系所單位： | 醫學檢驗暨生物技術學系 | |
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