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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊文欽(Wen-Chin Yang) | |
| dc.contributor.author | Kai-Wei Chen | en |
| dc.contributor.author | 陳凱偉 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:28:31Z | - |
| dc.date.copyright | 2010-02-11 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-01-29 | |
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Contributions of human tumor xenografts to anticancer drug development. Cancer Res 2006; 66: 3351-4, discussion 4. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22796 | - |
| dc.description.abstract | 乳癌是全球女性主要致死的惡性腫瘤，在過去25年來，約70 %的抗癌藥物來自天然物或其衍生物。依賴現有抗癌化學藥物常引發乳癌細胞對化學藥物產生抗性，以及新型抗癌化合物的開發作為組合性治療有助於乳癌的治療。因此找尋不同抗癌機制的藥物有其必要性。先前的研究發現大花咸豐草的萃取物具有抗癌活性。大花咸豐草可作為食物或民俗草藥使用，可治療許多疾病。我們與其他實驗室發現，大花咸豐草中有一個咸豐草聚乙炔去糖化合物 (cytopiloyne aglycone，CPA)，具有抑制血管新生的生物活性。本論文中，我們首先證實CPA會隨著濃度增加，顯著地減少三種乳腺癌細胞 (TS/A、MCF-7和MDA-MB-231)的細胞存活率，其IC50值分別是3.12、3.37 和 3.56 μg/ml。顯示CPA對於乳癌細胞有良好的抑制生長能力，而且此抑制與雌激素的作用無關。進一步探討CPA的抗癌作用發現，低劑量CPA具有抑制細胞增生的能力，並且其抑制作用是長時間且癌細胞無法回復、消除，高劑量CPA會隨時間增加而導致癌細胞的死亡。流式細胞儀的分析顯示，CPA能引發TS/A乳癌細胞進行細胞凋亡，並且活化caspase 3/7的活性，造成TS/A乳癌細胞的死亡。利用TS/A乳腺癌荷瘤小鼠為動物模式，來評估CPA的抗腫瘤藥效。經給予TS/A荷瘤小鼠PTX和不同濃度的CPA，我們發現小鼠的腫瘤大小、重量都顯著地受到抑制，而小鼠本身並無出現嚴重的藥物毒性。免疫組織染色方式也證實，相較於控制組，CPA可抑制腫瘤細胞增生，促進腫瘤細胞凋亡，並且可以增加荷瘤小鼠的存活率。CPA在活體內的作用與體外細胞的作用一致。我們也發現PTX或CPA處理的荷瘤小鼠中，其骨髓與脾臟內的骨髓衍生抑制細胞 (MDSC)數量比控制組少，這結果顯示MDSC沒有上升可能是因為CPA抑制腫瘤細胞的生長和促進死亡，使得腫瘤誘發MDSC的能力下降所致。所有結果證明CPA在體內與體外都具有抑制乳腺癌細胞的能力，此一抑制作用可能是透過抑制細胞增生和促進癌細胞凋亡之途徑。 | zh_TW |
| dc.description.abstract | Breast cancer is a leading cause of death in women worldwide. For the past 25 years, 70 % of anticancer drugs have been developed from natural products or their derivatives. However, a major obstacle to a successful cancer therapy is the problems of drug resistance in cancer patients. Therefore, there is a need to seek new anti-cancer drugs with different mechanisms. Bidens pilosa is commonly used as a food or folk medicines for different diseases. We and other groups have identified cytopiloyne aglycone (CPA) from B. pilosa. CPA was reported to inhibit angiogenesis in human umbilical vein endothelial cells (HUVEC). However, the anti-tumor effects and mechanisms of CPA are poorly understood. In this study, we first investigated the effect of CPA on the cell growth of TS/A, MCF-7 and MDA-MB-231 cells. We found that CPA inhibited tumor cell growth in a dose dependent manner. The IC50 value for TS/A, MCF-7 and MDA-MB-231 cells were 3.12, 3.37 and 3.56 μg/ml, respectively. Furthermore, low dose of CPA inhibited TS/A cell proliferation and the effect on anti-proliferation was persistent and irreversible. High dose of CPA have cell cytotoxity to induce apoptosis of TS/A cells, as revealed by induce the caspase 3/7 activity in flow-cytometer data.
We assessed anti-tumor effect of CPA in TS/A tumor-bearing BALB/c mice. We found that CPA treatment at 0.02, 0.1, 0.5 mg/kg suppressed the growth of TS/A tumors compared with control group mice, as evidenced by tumor size and mass via 24 days treatment period. And mice did not show serious symptoms of toxicity. The immunohistochemical staining results showed that the tumors from CPA-treated mice exhibited reduced cell proliferation and increased apoptosis compared with tumor from control group mice. Finally, the survival rate of any CPA-treated groups was prolonged significantly. The MDSC of bone marrow and spleen from PTX and CPA-treated mice were reduced. In conclusion, CPA had the effect of anti-tumor in breast cancer both in-vitro and in-vivo with reduced cell proliferation and induced cell apoptosis. There studies of CPA for chemotherapy medicine against breast cancer are thus warranted. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:28:31Z (GMT). No. of bitstreams: 1 ntu-99-R96b41011-1.pdf: 1825519 bytes, checksum: 7ae0e8369c2642c6b76e484d320a078f (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 目 錄
口試委員會審定書 ------------------------------------------------------------------------------- I 致謝 ------------------------------------------------------------------------------------------------ II 中文摘要 -------------------------------------------------------------------------------------- III 英文摘要 ---------------------------------------------------------------------------------------- V 目錄 --------------------------------------------------------------------------------------- VII 圖目錄 --------------------------------------------------------------------------------------- XII 表目錄 --------------------------------------------------------------------------------------- XIII 第一章、前言 -------------------------------------------------------------------------------------- 1 第一節、乳癌介紹與臨床治療方法 ---------------------------------------------------- 2 第二節、MDSC (myeloid-derived suppressor cells)介紹 --------------------------- 4 第三節、抗癌天然藥物之發展 ----------------------------------------------- 5 第四節、大花咸豐草(Bidens pilosa L.) 與cytopiloyne aglycone (CPA) ---------- 5 第二章、實驗材料 ------------------------------------------------------------------------------- 8 第一節、細胞株與細胞培養 ------------------------------------------------------------ 9 第二節、抗體 ------------------------------------------------------------------------- 10 第三節、藥品試劑 ----------------------------------------------------------------------- 10 第四節、緩衝液配方 --------------------------------------------------------------------- 10 第五節、其他實驗耗材 ------------------------------------------------------------------ 12 第六節、實驗儀器與分析軟體 -------------------------------------------------------- 12 第三章、實驗架構與方法----------------------------------------------------------------------- 13 第一節、實驗架構 ------------------------------------------------------------------------ 14 第二節、實驗方法 --------------------------------------------------------------- 14 一、細胞存活率分析 (Cell viability assay) ------------------------------------- 14 二、TS/A經過CPA作用24小時，移除，繼續測定細胞的存活率 ------- 14 三、低劑量CPA處理TS/A乳癌細胞24、48小時，計算細胞的死活 --- 14 四、細胞凋亡分析 (Annexin V & PI staining) ------------------------------- 15 五、PhiPhiLux測定caspase 3/7活性 -------------------------------------------- 15 六、以TS/A乳癌腫瘤負荷小鼠模式評估CPA抗腫瘤能力------------------ 15 七、TS/A腫瘤組織染色------------------------------------------------------------- 16 八、紀錄TS/A乳癌腫瘤負荷小鼠的存活率 (survival rate) ----------------- 17 九、荷瘤小鼠的骨髓和脾臟製備與流式細胞儀分析MDSC --------------- 17 十、統計與數據分析 --------------------------------------------------------------- 18 第四章、實驗結果 ------------------------------------------------------------------------------ 19 第一節、CPA的處理降低不同類型乳癌細胞的存活率 -------------------------- 20 第二節、低劑量CPA抑制細胞增生、高劑量CPA造成細胞死亡之藥物特性- 20 第三節、CPA引發TS/A乳癌細胞凋亡(apooptosis)，活化、增加caspase 3/7的活性 ------------------------------------------------------------------------------------- 21 第四節、給予CPA處理能削弱TS/A乳癌負荷小鼠的腫瘤大小、重量，抑制腫瘤成長 ---------------------------------------------------------------------------------- 22 第五節、CPA處理導致TS/A乳癌負荷小鼠的腫瘤，細胞增生的降低以及提高細胞凋亡的比例--------------------------------------------------------------------------- 23 第六節、CPA能延長TS/A乳癌負荷小鼠的存活率與壽命 ---------------------- 23 第七節、CPA能抑制TS/A腫瘤誘導的MDSC比例增加 ------------------------ 24 第五章、實驗討論 ------------------------------------------------------------------------------ 25 第一節、CPA具有廣泛地抑制多種乳癌細胞，有可能影響細胞週期，進而抑制乳癌細胞無限制的增生 ------------------------------------------------------------- 26 第二節、CPA促進乳癌細胞進行細胞凋亡，導致癌細胞的死亡 ---------------- 26 第三節、CPA有很好的抑制活體內腫瘤生長的效果，並且無副作用、能延長小鼠存活壽命，減少MDSC的累積 ------------------------------------------------- 27 第四節、總結 ------------------------------------------------------------------------------ 28 第六章、圖表 ------------------------------------------------------------------------------------ 29 第七章、參考文獻 ------------------------------------------------------------------------------ 47 圖目錄 圖一、Picture of Bidens pilosa and chemical structure of cytopiloyne aglycone ------- 31 圖二、Effect of CPA on cell viability of three different mammary gland cell lines ---- 32 圖三、Pulse-exposure effect of CPA on TS/A cells ----------------------------------------- 34 圖四、Effect of CPA on TS/A cell growth --------------------------------------------------- 35 圖五、CPA induces apoptosis in TS/A cells ------------------------------------------------- 36 圖六、Effect of CPA on the activation of caspases 3/7 ------------------------------------- 37 圖七、Effect of CPA on tumor growth in TS/A tumor-bearing mice --------------------- 40 圖八、Immunohistochemical analysis of TS/A tumors ------------------------------------- 42 圖九、Effect of CPA on the survival rate of TS/A tumor-bearing mice ------------------ 43 圖十、Effect of CPA on bone marrow and splenic MDSC in TS/A tumor-bearing mice --------------------------------------------------------------------------------------------- 45 附圖一、The suppressive functions of MDSC in tumor-bearing patients --------------- 46 表目錄 表一、Anti-cancer drugs and plant sources --------------------------------------------------- 30 表二、IC50 value of CPA in three different mammary gland cell lines ------------------- 33 | |
| dc.language.iso | zh-TW | |
| dc.subject | 免疫組織染色 | zh_TW |
| dc.subject | 抗癌天然物 | zh_TW |
| dc.subject | 骨髓衍生抑制細胞 | zh_TW |
| dc.subject | 大花咸豐草 | zh_TW |
| dc.subject | 咸豐草聚乙炔去糖化合物 | zh_TW |
| dc.subject | 抗細胞增生與細胞凋亡 | zh_TW |
| dc.subject | TS/A乳腺癌荷瘤小鼠 | zh_TW |
| dc.subject | Myeloid-Derived Suppressor Cells (MDSC) | en |
| dc.subject | Anti-cancer natural products | en |
| dc.subject | Bidens pilosa | en |
| dc.subject | Cytopiloyne aglycone | en |
| dc.subject | Anti-proliferation and apoptosis | en |
| dc.subject | TS/A tumor-bearing mice | en |
| dc.subject | Immunohistochemical staining | en |
| dc.title | 評估cytopiloyne aglycone在TS/A乳腺癌荷瘤小鼠的抗腫瘤效果與機制 | zh_TW |
| dc.title | Anti-tumor effect and mechanism of cytopiloyne aglycone in TS/A mammary gland tumor-bearing mice | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 顧記華(Jih-Hwa Guh),潘建源(Chien-Yuan Pan) | |
| dc.subject.keyword | 抗癌天然物,大花咸豐草,咸豐草聚乙炔去糖化合物,抗細胞增生與細胞凋亡,TS/A乳腺癌荷瘤小鼠,免疫組織染色,骨髓衍生抑制細胞, | zh_TW |
| dc.subject.keyword | Anti-cancer natural products,Bidens pilosa,Cytopiloyne aglycone,Anti-proliferation and apoptosis,TS/A tumor-bearing mice,Immunohistochemical staining,Myeloid-Derived Suppressor Cells (MDSC), | en |
| dc.relation.page | 55 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2010-01-29 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-99-1.pdf 未授權公開取用 | 1.78 MB | Adobe PDF |
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