博碩士論文 105826014 詳細資訊




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姓名 蕭升揚(Sheng-Yang Hsiao)  查詢紙本館藏   畢業系所 系統生物與生物資訊研究所
論文名稱 傳統中藥複方FY001W是三陰性乳腺癌新型的候選藥物
(Traditional Chinese Medicine FY001W as a novel candidate formula for triple negative breast cancer)
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摘要(中) 乳腺癌是全世界女性最常見的癌症之一且有很多的亞型,而三陰性乳腺癌是乳腺癌中死亡率最高、治療後的預後效果最差的亞型。目前治療方式為放射治療、化學治療及乳房切除,這些治療的方式都會造成嚴重的副作用。中草藥的治療已經在中國有多年的記載並且文獻指出有較低的副作用及毒性。首先,我們選擇的古書為普濟方,此書為一本最完整且收入最多方劑的古書。從書中分析乳石門的複方並利用卡方檢定,篩選出升麻湯 (FY001W) 且可以有效地促使細胞的凋亡。而在傳統複方中有君、臣、佐、使的分類可以協助並了解複方中的核心藥材。利用統計方式分別找出4種加減方,並探討其加減方強弱相對於正常細胞及三陰性乳腺癌癌細胞的毒性的差異,從中找出FY003W可能為其核心藥材。利用流式細胞儀檢視出FY001W及其加減方對於三陰性乳腺癌細胞促使細胞走向凋亡,並且利用西方墨點法不同藥物不同濃度處理後觀察蛋白質的表現量,發現casepase-3、caspase-7、PARP隨著濃度上升表現量隨之降低,表示FY001W及其加減方能使細胞走向內部凋亡,而為了瞭解低濃度時細胞的存活我們查看了自噬作用的蛋白標誌ATG5、ATG16L1、ATG7、LC3A/B,發現隨著濃度上升其表現量一致地下降代表自噬作用的效果降低。未來我們會增加分析的背景資料並強化過去的資料,希望能更有效地找尋乳石門中的核心藥材。
摘要(英) Breast cancer is one of the most common cancers among women worldwide and has many subtypes. Triple-negative breast cancer is the subtype with the highest mortality in breast cancer and the worst prognosis after treatment. Current treatments are radiation therapy, chemotherapy, and mastectomy, all of which can cause serious side effects. The Chinese Medicine has documented for many years in China and the paper indicates that it has low side effects and toxicity. In our study, we find an ancient book, Pu Ji Fang, which is the most complete and comprehensive. We analyze formula of Ru Shi Men from Pu Ji Fang. Using the chi-square test to screen Sheng Ma Tang (FY001W) and effectively promote triple negative breast cancer cells apoptosis. However, there are consist of monarchs, ministers, Zuo, and ambassadors in the traditional formula. This classification assist and understand the core herbs in the formula. We hope to find the core herbs so using statistics to identify four derivatives. The FY001 and derivatives were treated in the triple negative breast cancer cells to induced apoptosis by flow cytometry, and the expression of protein were observed the different concentrations of different drugs by western blot. It was found that the protein expression casepase-3, caspase-7 and PARP decreased with the increase of concentration, indicating the cells underwent apoptosis through the intrinsic pathway. In order to realize the survival of cells at low concentrations, we investigate the autophagy markers ATG5, ATG16L1, ATG7, and LC3A/B. Finding the consistent decrease in the expression level as the concentration increased indicates that the effect of autophagy was reduced. In the future, we will enrich the background information to analyze and hoping to find the core herbs.
關鍵字(中) ★ 乳癌
★ MDA-MB-231
★ 凋亡
★ 自噬作用
★ 中草藥
★ 複方
★ 升麻湯
關鍵字(英) ★ breast cancer
★ MDA-MB-231
★ apoptosis
★ autophagy
★ TCM
★ formula
論文目次 中文摘要 I
Abstract II
致謝 III
圖目錄 VI
表目錄 VII
補充圖目錄 VIII
補充表目錄 IX
緒論 1
1-1 乳癌 1
1-2 乳腺癌的分群和靶向治療 2
1-3 治療及中藥的好處 3
1-4 普濟方 4
1-5 自噬作用及凋亡作用 5
1-6 研究目的 7
實驗方法與材料方法 8
2-1 細胞培養 8
2-2 普濟方資料整理 8
2-3 中草藥的配方 9
2-4 傳統中藥的萃取 10
2-5 細胞存活測定 11
2-6 藉由Flow檢測細胞的凋亡 11
2-7 蛋白質萃取 12
2-8 西方墨點法 12
2-9 晶片生物訊息分析 13
結果 14
3-1 生物資訊分析及小分子藥物 14
3-1-1 生物晶片 14
3-1-2 生物信息分析 16
3-1-3 nonoxynol-9 17
3-2 傳統中藥高通量藥物篩選系統 18
3-3 古書的探討及分析 20
3-3-1 普濟方的分析 20
3-3-2 藉由Alamar Blue研究FY001的毒性 29
3-3-3 藉由流式細胞儀凋亡的分析 30
3-4 FY001W加減方之療效 33
3-4-1 傳統中藥複方用藥配伍 33
3-4-2 FY001W加減方FY002及FY003誘導乳腺癌細胞凋亡 37
3-4-3 FY001W加減方FY004及FY005誘導乳腺癌細胞凋亡 41
3-4-4 FY001W及其加減方之藥效 42
3-5 FY001W及FY001W的加減方訊息途徑分析 44
討論 49
參考文獻 51
補充資料 53
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指導教授 蘇立仁(Li-Jen Su) 審核日期 2019-1-29
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