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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 蔣丙煌 | |
| dc.contributor.author | Yu-Ting Huang | en |
| dc.contributor.author | 黃鈺婷 | zh_TW |
| dc.date.accessioned | 2021-06-15T11:42:44Z | - |
| dc.date.available | 2021-10-05 | |
| dc.date.copyright | 2016-10-05 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-15 | |
| dc.identifier.citation | Aggarwal, B. B., et al. (2003). Anticancer potential of curcumin: preclinical and clinical studies. Anticancer Research, 23(1A), 363-398.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49699 | - |
| dc.description.abstract | 國人十大死因之首為癌症，其中大腸結直腸癌(colorectal cancer, CRC)位居第三，與國人之飲食、生活習慣、遺傳等因素相關。過去研究指出微環境改變和癌症幹細胞存在對癌症發生、轉移與再發具有密不可分之關係。本研究目的為探討植化素與微環境調控對大腸直腸癌癌症幹細胞(colorectal cancer stem cells，CCSCs)發展之影響。實驗建立以CRC病患之正常、瘜肉與癌組織分析CCSCs marker (CD133與CD44)之表現量及代謝體，接續建立CRC初代細胞培養平台，觀察植化素對各CRC初代細胞中CCSCs及代謝體之影響，再者以CRC細胞株HCT-116、HT-29、SW480及SW620細胞株中CCSCs表現分析及微環境因子對CCSCs發展之影響，以CCSCs表現量高低，模擬CRC細胞及瘜肉細胞，針對CCSCs高度表現之HT-29細胞株給予植化素curcumin (Cur)、resveratrol (Res)及4-Acetylantroquinonol B (4AAQB)探討其對細胞存活率、凋亡、CCSCs表現的影響，而CCSCs表現量低之SW480細胞株則給予不同微環境因子如reactive oxygen species (ROS)、epidermal growth factor (EGF)及transforming growth factor (TGF)誘導CCSCs表現。實驗結果發現CRC腺瘤性息肉(N=61)的代謝特徵相較於CRC（N = 57）組織，其磷酸腺苷（Adenosine monophosphate, AMP），adenine，5'- methythioadenosine，3-hydroxybutyric acid ，prostaglandin E2， threonine 和 glutamine具統計上顯著差異（P <0.05）。因CRC初代細胞培養平台建置尚未完成，且以微環境因子誘導CRC細胞株表現CCSCs部分，發現以單一因子無法有效誘導CCSCs表現，因此以CRC細胞株進行研究。結果發現以50 μM curcumin處理48小時確實增加CRC及CCSCs的細胞凋亡，但僅在CD44+細胞具顯著性。在CRC細胞株的代謝特徵的研究，curcumin處理後CD44-細胞中glyceraldehyde和hydroxypropionic acid之含量增加，但以curcumin處理的CRC和CD44+細胞則glutamine含量減少。基於我們的人體組織和癌細胞株的代謝概況比較，我們認為，curcumin-CD44+耦合在細胞膜上可能對glutamine的運輸產生某些阻斷作用，抑制其進入細胞內，因此降低了glutamine在CD44+細胞中的含量以至於誘導細胞凋亡的發生。 | zh_TW |
| dc.description.abstract | Colorectal cancer (CRC) is the three major cause of cancer-related mortality in Taiwan. The risk factors of CRC include age, life style and family history. This study investigated the effect of microenvironment and phytochemicals on the development of colorectal cancer stem cells (CCSCs) and its possible mechanism. We analyzed CRC patient’s normal, adenomatous polyps and tumor tissue for expression of CCSCs (CD133 and CD44) and metabolic profiles. We also tried to establish CRC primary cells for investigating the effect of phytochemicals and microenvironmental factors on the CCSCs and metabolic profiles. In addition, the microenviroments factors including reactive oxygen species (ROS)、epidermal growth factor (EGF)及transforming growth factor (TGF) were used to induce the expression of CCSCs. However, both of the attempt of establishing CRC primary culture and induction of CCSCs by microenvironmental factors failed, we decided to use CRC cell lines (HCT-116, HT-29, SW480 and SW620) model to analyze their expression of CCSCs and metabolic profiles. Comparison of the metabolic profiles of human adenomatous polyp (N=61) and colorectal cancer (CRC) (N=57) tissue found statistically significant differences (p<0.05) in their composition of Adenosine monophosphate (AMP), adenine, 5’-methythioadenosine, 3-hydroxybutyric acid, prostaglandin E2, threonine and glutamine. Our cell culture model study found that curcumin treatment (50 μM for 48 h) did increased apoptosis of CRC cells and CCSCs, but only the CD44+ cells reached statistically significant. Further metabolic profile studies of the CRC, CD44+ and CD44- cells indicated that curcumin treatment increased glyceraldehyde and hydroxypropionic acid in CD44- cells, but decreased glutamine content in both curcumin-treated CRC and CD44+ cells. Based on the comparison of the metabolic profiles of human tissues and cancer cells we suggest that curcumin might couple with CD44 and that curcumin-CD44+ coupling at the cell membrane might have some blocking effect on the transport of glutamine into the cells, thus decreasing the glutamine content in the CD44+ cells and inducing apoptosis. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T11:42:44Z (GMT). No. of bitstreams: 1 ntu-105-D98641003-1.pdf: 5950771 bytes, checksum: 493d29d84b058e9854924b3baa6418a9 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 謝誌 i
摘要 iii Abstract v 目錄 I 圖目錄 IV 表目錄 VII 壹、 前言及研究目的 1 研究目的 2 貳、 文獻回顧 3 第一章、 大腸結直腸癌 (Colorectal cancer, CRC) 3 1.1 大腸結直腸癌的風險因子 4 1.2 大腸結直腸癌的分子發病學與致病機轉 9 第二章、 癌症幹細胞 (Cancer stem cell, CSC) 19 2.1 癌症幹細胞的特徵與來源 23 2.2 癌症幹細胞的分離與檢測方法 32 第三章、 癌症細胞微環境 (Microenvironment of cancer cells) 37 3.1 癌症細胞微環境介紹 37 3.2 癌症微環境與腫瘤發展之相關 40 第四章、 植化素 (Phytochemicals) 42 4.1 植化素的簡介 42 4.2 植化素的生理活性與選擇 43 第五章、 代謝體 (Metabolomics) 46 第六章、 腫瘤細胞代謝之改變 49 第七章、 實驗架構 54 7.1 實驗構想 54 7.2 人體大腸結直腸正常組織、大腸結直腸癌瘜肉與大腸結直腸癌組織CCSCs marker表現量及代謝體分析 55 7.3 建立人體大腸結直腸正常組織、大腸結直腸癌瘜肉與大腸結直腸癌組織初代細胞培養平台，觀察植化素對各人體組織初代細胞CCSCs marker表現量及代謝體之影響 56 7.4 人類大腸結直腸癌細胞株HCT-116、HT-29、SW480及SW620癌症幹細胞表現分析 57 7.5 觀察CRC細胞株(A) 高CCSCs marker表現量細胞株及(B)低CCSCs marker表現量細胞株，經微環境因子調控誘導CCSCs marker表現，植化素介入對(A)(B)兩種CRC細胞株之CCSCs marker表現量、細胞存活率、細胞凋亡及細胞株內代謝體之影響 58 參、 材料與方法 60 第一章、 實驗材料與儀器設備 60 1.1 細胞株來源 60 1.2 藥品試劑 60 1.3 儀器設備 63 第二章、 實驗方法 64 2.1 樣品配置 64 2.2 細胞培養 64 2.3 細胞存活率分析 65 2.4 蛋白質萃取與定量 67 2.5 西方墨點法 69 2.6 以流式細胞儀分析癌症幹細胞表現量 73 2.7 以微環境因子誘導癌症幹細胞表現 75 2.8 細胞凋亡(Annexin V/PI double staining) 76 2.9 代謝體分析 77 第三章、 統計分析 78 肆、 結果與討論 79 第一章、 人類大腸結直腸正常組織、大腸結直腸癌瘜肉與大腸直腸癌癌組織CCSCs marker表現及代謝體分析 79 第二章、 建立人體大腸結直腸正常組織、大腸直結腸癌瘜肉組織與大腸結直腸癌組織初代細胞培養平台，觀察植化素對各人體組織初代細胞CCSCs marker表現量及代謝體之影響 85 第三章、 人類大腸結直腸癌細胞株HCT-116、HT-29、SW480及SW620癌症幹細胞表現分析 90 第四章、 植化素介入對HT-29及經微環境調控誘導後SW480兩種CRC細胞株之CCSCs marker表現量、存活率、凋亡及代謝體之影響 97 第五章、 植化素之給予對人類大腸結直腸癌細胞株HT-29細胞凋亡表現之影響 111 伍、 結論 119 陸、 參考文獻 120 柒、 附錄 135 | |
| dc.language.iso | zh-TW | |
| dc.subject | apoptosis | zh_TW |
| dc.subject | 大腸結直腸癌 | zh_TW |
| dc.subject | 癌症幹細胞 | zh_TW |
| dc.subject | glutamine | zh_TW |
| dc.subject | curcumin | zh_TW |
| dc.subject | glutamine | en |
| dc.subject | curcumin | en |
| dc.subject | apoptosis | en |
| dc.subject | cancer stem cell | en |
| dc.subject | colorectal cancer | en |
| dc.title | 探討微環境及植化素對大腸直腸癌癌症幹細胞發展之影響 | zh_TW |
| dc.title | Effect of microenvironment and phytochemicals on the development of colorectal cancer stem cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 潘敏雄,鍾景光,潘子明,邱瀚模 | |
| dc.subject.keyword | 大腸結直腸癌,癌症幹細胞,apoptosis,curcumin,glutamine, | zh_TW |
| dc.subject.keyword | colorectal cancer,cancer stem cell,apoptosis,curcumin,glutamine, | en |
| dc.relation.page | 161 | |
| dc.identifier.doi | 10.6342/NTU201602520 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-08-15 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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