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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄭光成(Kuan-Chen Cheng) | |
| dc.contributor.author | Yen-Yi Lin | en |
| dc.contributor.author | 林彥沂 | zh_TW |
| dc.date.accessioned | 2021-06-08T02:50:19Z | - |
| dc.date.copyright | 2017-08-24 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-08-16 | |
| dc.identifier.citation | 香港癌症基金會 (2015) 認識癌症:前列腺癌.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/20484 | - |
| dc.description.abstract | 本研究透過醱酵的方式來生產臺灣特有種靈芝，臺灣紫芝 (Ganoderma formosanum) 的菌絲體，並先以水以及酒精兩種溶劑對菌絲體進行萃取，並測試其萃取物對於前列腺癌細胞的細胞毒性。結果顯示，臺灣紫芝菌絲體酒精萃取物 (Ganoderma formosanum ethanolic extract, GF-E) 能夠展現出比水萃取物 (Ganoderma formosanum aquatic extract, GF-A) 更為有效的抑制作用，進一步以四個不同極性的有機溶劑將GF-E以液相分配 (liquid-liquid partition) 的方式進行進一步的純化，結果顯示GF-E在以己烷 (GF-E hexane partitioned fraction, GF-EH) 以及丁醇 (GF-E butanol partitioned fraction,GF-EB) 進行純化後得到的萃取層GF-EH以及GF-EB對於癌細胞增生的抑制效果最佳。接著以即時聚合酶連鎖反應、西方墨點法以及流式細胞儀進行分析後發現，GF-EH能抑制前列腺癌細胞DU145的細胞週期，使其停滯在G1期；GF-EB則能誘導DU145，發生細胞凋亡，分別透過兩種不同的機制來抑制前列腺癌細胞的增生。結果顯示臺灣紫芝菌絲體的萃取物，具應用在治療前列腺癌的潛力。 | zh_TW |
| dc.description.abstract | In this study, fermentation was employed to produce Ganoderma formosanum mycelia. Ganoderma formosanum is an endemic Ganoderma species of Taiwan. Then bioactive metabolites with anti-proliferation effect on prostate cancer cells in mycelia was isolated through extraction and tested the cytotoxic of the extracts. Results demonstrated that extracts from G. formosanum ethanolic extract(GF-E) exhibited superior cytotoxicity to cancer cells compared with aquatic extract(GF-H). After further purification, the results showed that hexane faction(GF-EH) and butanol fraction(GF-EB) exerted a significant cytotoxicity on DU145 cells. Furthermore, it was found that these fractions may exert anti-proliferation on prostate cancer cells via inhibiting cell cycle process, GF-EH arrested cells in G1 phase. On the other hand, GF-EB upregulated Caspase-3 responsible for apoptosis, leading to cell apoptosis. The present study indicated that the mycelia extracts of G. formosanum exerted anti-proliferation effect on prostate cancer cells, suggesting G. formosanum is a potential candidate to treat prostate cancer. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T02:50:19Z (GMT). No. of bitstreams: 1 ntu-106-R04642008-1.pdf: 3774093 bytes, checksum: f02e5cfe1f288eb4c83ea7c3734c2e8f (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 謝誌 i
摘要 ii Abstract iii 目錄 iv 圖目錄 viii 表目錄 x 壹、 前言 1 貳、 文獻回顧 2 第一節、 靈芝 2 1.1 靈芝分類 4 1.2靈芝培養 6 1.3生物活性 6 1.4生物活性成分 6 1.4.1靈芝三萜類 6 1.4.2靈芝多醣體 9 1.5臺灣紫芝 12 第二節、前列腺癌 14 2.1 前列腺癌 14 2.2 病患及死亡人數分布 15 2.3 前列腺癌的檢測以及治療方法 18 第三節、 靈芝抑癌機制 20 3.1 In vitro 試驗 20 3.1.1細胞週期 20 3.1.2細胞凋亡 22 3.1.3 NFkB，AP-1 24 3.2 In vivo 試驗 24 3.2.1免疫調節 25 3.2.2血管新生 25 參、 材料與方法 27 第一節、菌絲體培養 28 1.1 菌株 28 1.2 儀器設備 28 1.3 實驗藥品 29 1.4 培養方法 29 1.4.1平板培養 29 1.4.2液態培養 30 1.4.2.1種菌培養 30 1.4.2.2搖瓶培養 30 1.4.2.3生物反應器培養 30 1.5 培養基配置 31 第二節、 菌絲體萃取 32 2.1 儀器及藥品 32 2.2菌絲體乾燥 33 2.3菌絲體萃取 33 2.3.1 水萃取 33 2.3.2 酒精萃取 33 2.3.3液相分配 33 2.3.4 LH-20管柱純化 34 2.3.4.1 LH-20管柱填充 34 2.3.4.2 樣品純化 34 2.3.4.2 TLC分析 34 第三節、 細胞實驗 36 3.1 細胞株 36 3.2儀器及材料 36 3.3細胞培養 39 3.3.1 DU145細胞培養 39 3.3.2 LNCaP細胞培養 39 3.3.3 Fibroblast細胞培養 39 3.4細胞計數 39 3.5細胞毒性測試 40 3.6流式細胞儀分析細胞週期 40 3.6.1細胞加藥與染色 40 3.6.2細胞群圈選與DNA含量分析 41 3.7反轉錄聚合酶連鎖反應分析mRNA表現量變化 42 3.7.1抽RNA 42 3.7.2 RNA濃度定量 42 3.7.3 反轉錄cDNA 42 3.7.4 qRT-PCR 43 3.7.5瓊脂凝膠電泳 43 3.8西方墨點法分析蛋白質表現量變化 44 3.8.1萃取蛋白質 44 3.8.2蛋白質定量 44 3.8.3 SDS PAGE 44 3.8.3.1 膠體配置 44 3.8.3.2 鑄膠 45 3.8.3轉印 46 3.8.5免疫染色 46 3.8.6 抗體剝離 46 第四節、分析方法 47 肆、 結果與討論 48 第一節、 菌絲體液態深層培養 48 1.1 菌絲體培養 48 第二節、 菌絲體萃取物抑制前列腺癌細胞增生能力評估 50 2.1 萃取物對癌細胞DU145毒性評估 50 2.2 萃取溫度對於細胞毒性影響評估 58 2.3萃取物對癌細胞LNCaP毒性評估 61 2.4 萃取物對癌細胞Fibroblast毒性評估 64 第三節、細胞生長抑制路徑分析 67 3.1 DNA電泳結果分析DNA片斷化 67 3.2週期素以及凋亡蛋白酶表現變化分析 69 3.3流式細胞儀分析細胞週期 72 第四節、LH-20 純化結果 73 伍、結論與未來展望 77 陸、參考文獻 79 | |
| dc.language.iso | zh-TW | |
| dc.title | 臺灣紫芝萃取物抑制前列腺癌細胞增殖之活性評估 | zh_TW |
| dc.title | The Anti-Proliferation Activity of Ganoderma formosanum Extracts on Prostate Cancer Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 沈偉強(Wei-Chiang Shen),劉?睿(Je-Ruei Liu),劉啟德(Chi-Te Liu),陳宏彰(Hong-Jhang Chen) | |
| dc.subject.keyword | 臺灣紫芝,深層液態培養,萃取,細胞週期,前列腺癌, | zh_TW |
| dc.subject.keyword | Ganoderma formosanum,extracts,anti-proliferation,prostate cancer,cell cycle,apoptosis, | en |
| dc.relation.page | 116 | |
| dc.identifier.doi | 10.6342/NTU201703326 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2017-08-16 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物科技研究所 | zh_TW |
| 顯示於系所單位： | 生物科技研究所 | |
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