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研究生:古珮渝
研究生(外文):GU,PEI-YU
論文名稱:探討苦參根純化的苦參 G 誘導人類乳癌細胞之細胞凋亡的分子機制
論文名稱(外文):Sophoraflavanone G Isolated from The Root of Sophora flavescens Through Induced of Apoptosis in Human Breast Cancer Cells.
指導教授:黃文忠黃文忠引用關係
指導教授(外文):HUANG,WEN-CHUNG
口試委員:賴裕順劉倩君
口試委員(外文):LAI,YU-SHENLIOU,CHAIN-JIUN
口試日期:2017-06-28
學位類別:碩士
校院名稱:長庚科技大學
系所名稱:健康產業科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:中文
論文頁數:68
中文關鍵詞:苦參G細胞凋亡乳癌MDA-MB-231
外文關鍵詞:Sophoraflavanone GApoptosisBreast cancerMDA-MB-231
相關次數:
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  • 下載下載:1
  • 收藏至我的研究室書目清單書目收藏:0
苦參 G(SG;Sophoraflavanone G)萃取自中藥苦參。而本實驗室先前研究發現,在人類血癌細胞中, SG 能促使癌細胞走向凋亡,抑制血癌細胞的生長。因此,本研究則是探討 SG 對於 MDA-MB-231 乳癌細胞中的促凋亡現象及相關分子機制。 MDA-MB-231 細胞給予不同濃度的 SG 處理,使用 CCK-8 方法評估 MDA-MB-231 細胞的存活力,並也利用 DAPI 螢光染色觀察到細胞凋亡的核萎縮情形。同時也觀察了 SG 對 MDA-MB-231 細胞刺激後,細胞內 ROS 的產生和細胞週期的影響。從實驗結果得知,隨著給予刺激 SG 濃度增加,MDA-MB-231 細胞凋亡情況也隨之增加,同時細胞核萎縮情形也有顯著增加;另外,使用 Western 觀察細胞凋亡的相關蛋白,在蛋白質表現量的部分, SG 增加了乳癌細胞中細胞凋亡相關蛋白 caspase-8 和 caspase-9、caspase-3的表現量。SG 也促使細胞中 ROS 表現大量增加,以及造成 DNA 損傷,而細胞週期則增加 G0/G1 的停滯。從上述實驗結果發現,SG 通過 ROS ,caspase 8 和 caspase 9、caspase 3 活化來抑制 MDA-MB-231 細胞的生長並誘導細胞走向凋亡。因此,透過這些細胞實驗研究,SG 有希望的成為有效治療乳癌的植物純化物。
Sophoraflavanone G (SG) was isolated from Sophora flavescens, and our earlier studies demonstrated that SG is able to induced apoptosis in human leukemia cells. In this study, our aim evaluated the apoptosis effects of SG and its underlying molecular mechanism in breast cancer MDA-MB-231 cells. MDA-MB-231 cells treated with various concentrations of SG. The viability of MDA-MB-231 cells was measured using the CCK method, and nuclear condensation indicative of apoptosis was detected by DAPI fluorescence staining. Furthermore, apoptotic signal proteins were detected by Western blotting. We also observed ROS and cell cycle in SG treated with MDA-MB-231 cells. The results demonstrated that SG could induced significantly DNA fragmentation and nuclear condensation in MDA-MB-231 cells. SG also could activate caspase 8, caspase 9 and caspase 3 expression. We found that SG could induce ROS expression to induce DNA damage, and hold cell cycle in G0/G1 phase. These results suggested that SG exerts its growth inhibitory effects on MDA-MB-231 cells through ROS, caspase 8, caspase 9 and caspase 3 activation for induction of apoptosis. Therefore, SG offers promising chemotherapeuric potential to prevent cancers such as breast cancer.
目錄
長庚科技大學學位論文授權書
指導教授推薦書
口試委員會審定書
中文摘要
英文摘要
第一章 緒論
 第一節 前言
  壹、癌症
  貳、癌細胞
  叁、細胞遷徙
  肆、乳癌
  伍、乳癌細胞(Breast cancer cell)
  陸、細胞凋亡(Apoptosis)
  柒、氧化壓力(ROS;Reactive Oxygen Species)
  捌、苦參 G(SG;Sophoraflavanone G)
 第二節 假說與研究目的
  壹、研究動機
  貳、實驗目的
  叁、細胞實驗流程圖
第二章 實驗材料與方法
 第一節 實驗方法
  壹、MDA-MB-231 細胞株與細胞培養
  貳、試驗藥材 Sophoraflavanone G(SG)
  叁、細胞存活率檢測(Cell viability assays)
  肆、DAPI 染色
  伍、膠凝體電泳(Western blot analysis)
  陸、細胞流式儀分析(Flow cytometry)
   一、細胞週期染色(PI;Propidium Iodide stain)
  柒、活性氧化物測定(ROS;Reactive oxygen species)
  捌、細胞遷移實驗(Migration Assay)
  玖、DNA 片段(DNA fragmentation)
  拾、統計分析
第三章 結果
 第一節 苦參 G 對於 MAD-MB-231 細胞存活率影響
 第二節 苦參 G 對於 MAD-MB-231 細胞型態的影響
 第三節 苦參 G 對於 MAD-MB-231 細胞氧化壓力影響
 第四節 苦參 G 對於 MAD-MB-231 細胞週期的影響
 第五節 苦參 G 對於 MAD-MB-231 細胞 DNA 損傷之情況
 第六節 苦參 G 對於 MAD-MB-231 細胞遷徙之情況
 第七節 苦參 G 對於 MAD-MB-231 細胞凋亡相關蛋白之影響
第四章 討論
 第一節 細胞實驗
 第二節 苦參 G 對於 MAD-MB-231 細胞型態的影響
 第三節 苦參 G 對於 MAD-MB-231 細胞氧化壓力影響
 第四節 苦參 G 對於 MAD-MB-231 細胞週期的影響
 第五節 苦參 G 對於 MAD-MB-231 細胞 DNA 損傷之情況
 第六節 苦參 G 對於 MAD-MB-231 細胞遷徙之情況
 第七節 苦參 G 對於 MAD-MB-231 細胞凋亡相關蛋白之影響
第五章 結論
參考文獻
圖表

圖目錄
圖1 癌細胞的十大基本特徵
圖2 乳癌期別
圖3 乳房X光檢查方式模擬圖
圖4 乳腺管原位癌(DCIS;Ductal Carcinoma In Situ)
圖5 細胞凋亡路徑示意圖
圖6 ROS 的影響
圖7 苦參 G 分子結構
圖8 Sophoraflavanone G對MDA-MB-231細胞存活率影響
圖9 Sophoraflavanone G 對於 MAD-MB-231 細胞型態的影響
圖10 Sophoraflavanone G對於MAD-MB-231細胞氧化壓力影響
圖11 Sophoraflavanone G對於 MAD-MB-231 細胞週期的影響
圖12 Sophoraflavanone G對於 MAD-MB-231細胞 DNA 損傷之情況
圖13 Sophoraflavanone G對於 MAD-MB-231 細胞遷徙之情況
圖14 Sophoraflavanone G對於 MAD-MB-231 細胞凋亡相關蛋白之影響


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