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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 羅翊禎(Yi-Chen Lo) | |
dc.contributor.author | Pei-Shan Huang | en |
dc.contributor.author | 黃珮珊 | zh_TW |
dc.date.accessioned | 2021-07-10T21:33:38Z | - |
dc.date.available | 2021-07-10T21:33:38Z | - |
dc.date.copyright | 2017-02-21 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-02-15 | |
dc.identifier.citation | 1. Aparicio, T.; Baer, R.; Gautier, J., DNA double-strand break repair pathway choice and cancer. DNA Repair (Amst) 2014, 19, 169-75.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76600 | - |
dc.description.abstract | 人蔘皂苷Rg3是人蔘中具有生理活性的主要成分之一,多篇文獻指出其與抗癌
藥物一起使用可增加癌細胞對藥物的敏感性,並推測其機制可能為人蔘皂苷與藥 物會競爭運輸通道,造成抗癌藥物較無法被細胞排出。而在先前利用酵母菌進行 人蔘皂苷生物轉換的實驗中發現人蔘皂苷Rg3會進入酵母菌細胞內,因此利用酵母 菌特定運輸系統的單一剃除菌株分析細胞內人蔘皂苷Rg3含量,結果發現rgt1Δ突 變株細胞人蔘皂苷Rg3含量顯著低於野生型酵母菌;而pdr15Δ突變株細胞內人蔘皂苷Rg3含量則顯著高於野生型酵母菌株,故推測酵母菌會利用Rgt1蛋白進入胞內並透過Pdr15蛋白運輸至細胞外。因此,本實驗將利用Complementation確認這兩個基因調控並運輸人蔘皂苷Rg3進出細胞內外,但目前得到的質體皆無法正常表現蛋白。 此外,我們利用DNA修復缺失菌株:rad51Δ及rad52Δ突變株,分別與rgt1Δ及pdr15Δ突變株製作雙基因缺失菌株,探討人蔘皂苷Rg3對於這些菌株在藥物(MMS、Cisplatin)處理下的敏感性。結果發現:若將rad51Δ及rad52Δ突變株的PDR15基因剃除,其對於藥物的敏感度會提高,此外這些菌株若與人蔘皂苷Rg3共同培養後,其對於藥物的敏感度也會提高,然而利用UPLC-Q-TOF-MS/MS分析這些菌株細胞內人蔘皂苷含量並沒有看到顯著差異。而我們進一步想要了解是否能從市售產品攝取到人蔘皂苷Rg3,結果發現其內主要的人蔘皂苷為大分子結構,但都含有一定量的人蔘皂苷Rg3,此外並沒有看到分子結構更小的人蔘皂苷。 | zh_TW |
dc.description.abstract | Ginsenoside Rg3 is one of the primary pharmacologically active components in Panax ginseng. Previous studies showed that ginsenoside Rg3 is an effective agent to sensitize the growth of cancer cells in cancer chemotherapy. Studies also suggested that Rg3 bound to ABC transporters, thereby blocked drug efflux and resulted in the increased drug sensitivities in multidrug resistant cancer cells. Interestingly, our
previous study found that Rg3 was possibly transported into cells through glucose transporters and out of cells through ABC transporters in yeast, Saccharomyces cerevisiae. Thus, in this study is to further confirm ginsenosides Rg3 can be transported through RGT1 and PDR15 in yeast, however, we didn’t successfully construct expression plasmids. In addition, DNA-repair deleted strains will be treated with DNA damage agents, such as MMS and cisplatin combined with or without Rg3 to investigate if Rg3 transportation could affect drug sensitivities in yeast. Our results show rad51Δpdr15Δ and rad52Δpdr15Δ double mutant are more sensitive to anti-cancer drug, such as MMS and Cisplatin. Besides, ginsenoside Rg3 can also enhance the sensitivities to MMS and Cisplatin of these two double mutants. Hopefully, the results can provide more information about the transportation of ginsenosides and the possible future medical application of ginsenosides. | en |
dc.description.provenance | Made available in DSpace on 2021-07-10T21:33:38Z (GMT). No. of bitstreams: 1 ntu-106-R03641021-1.pdf: 14092018 bytes, checksum: f8488e9eeaaef7f4da671b94e13fb451 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 摘要 I
Abstract II 目錄 III 圖目錄 VI 表目錄 VIII 文獻回顧 1 1.1 人蔘皂苷 1 1.2 人蔘皂苷的抗癌活性 4 1.3 人蔘皂苷Rg3 5 1.4 人蔘皂苷Rg3的抗癌活性 7 1.5 人蔘皂苷Rg3藥物動力學 8 1.6 人蔘皂苷Rg3與癌症藥物的協同作用 8 1.7 人蔘皂苷Rg3調節癌細胞之多重抗藥性 9 1.8 酵母菌模式 10 1.9 人蔘皂苷Rg3的定性及定量分析 10 1.10 酵母菌運輸系統 12 1.11 酵母菌RGT1基因 13 1.12 酵母菌PDR5基因、PDR15基因 15 1.13 DNA修復缺失菌株 17 2 研究目的與實驗架構 19 3 材料與方法 20 3.1 實驗材料 20 3.1.1 酵母菌菌株 20 3.1.2 人蔘皂苷 20 3.1.3 市售人蔘產品 21 3.2 實驗藥品 21 3.2.1 化學試劑 21 3.2.2 微生物培養材料 21 3.3 儀器設備 22 3.3.1 實驗耗材 22 3.3.2 一般儀器設備 22 3.3.3 化學分析儀器設備 23 3.4 實驗方法 23 3.4.1 酵母菌培養條件 23 3.4.2 培養基配方 24 3.4.3 酵母菌生長曲線測量 24 3.4.4 轉殖方法 25 3.4.5 西方墨點法 28 3.4.6 市售人蔘產品萃取人蔘皂苷之方法 28 3.4.7 酵母菌細胞內及細胞外人蔘皂苷發酵液的收集及固相萃取 29 3.4.8 人蔘皂苷超高效液相層析質譜儀分析條件 29 3.4.9 高效液相層析串聯質譜之人蔘皂苷相對濃度計算 30 3.4.10 點試驗 Spotting Assay 30 3.4.11 酵母菌存活數試驗 32 3.4.12 統計分析 32 4 結果與討論 33 4.1 人蔘皂苷對酵母菌生長的影響 33 4.2 探討RGT1及PDR15基因剃除對與DNA損傷相關基因缺失菌株的影響 35 4.3 PDR15基因剃除調節rad51Δ及rad52Δ突變株對於MMS的敏感性 38 4.4 人蔘皂苷Rg3調節rad51Δ及rad52Δ突變株在MMS下的存活率 41 4.5 人蔘皂苷Rg3調節rad51Δ及rad52Δ突變株對於Cisplatin的敏感性 43 4.6 人蔘皂苷Rg3調節rad51Δ及rad52Δ突變株在Cisplatin下的存活率 46 4.7 比較酵母菌株細胞內外人蔘皂苷Rg3的含量 48 4.8 RGT1及PDR15質體轉殖與蛋白表現 50 4.9 市售產品的人蔘皂苷種類分析 62 5 結論 71 6 參考文獻 72 | |
dc.language.iso | zh-TW | |
dc.title | 探討運輸蛋白Pdr15 調節人蔘皂苷Rg3對基因修復缺失菌株在MMS 及Cisplatin 下的反應 | zh_TW |
dc.title | ABC Transporter Pdr15 regulates ginsenoside Rg3 to affect survival of DNA-repair mutants under MMS and Cisplatin treatments. | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 高承福,謝淑貞 | |
dc.subject.keyword | Pdr15,人蔘皂?,Rg3,基因修復缺失菌,MMS,cisplatin,Rgt1,RAD51,RAD52, | zh_TW |
dc.subject.keyword | ABC transporter,ginsenoside,Rg3,DNA-repair deleted strains,MMS,cisplatin,glucose transporter,RGT1,PDR15,RAD51,RAD52, | en |
dc.relation.page | 91 | |
dc.identifier.doi | 10.6342/NTU201700600 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2017-02-15 | |
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 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-106-R03641021-1.pdf 目前未授權公開取用 | 13.76 MB | Adobe PDF |
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