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標題: | 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 ABC Transporter Pdr15 regulates ginsenoside Rg3 to affect survival of DNA-repair mutants under MMS and Cisplatin treatments. |
作者: | Pei-Shan Huang 黃珮珊 |
指導教授: | 羅翊禎(Yi-Chen Lo) |
關鍵字: | Pdr15,人蔘皂?,Rg3,基因修復缺失菌,MMS,cisplatin,Rgt1,RAD51,RAD52, ABC transporter,ginsenoside,Rg3,DNA-repair deleted strains,MMS,cisplatin,glucose transporter,RGT1,PDR15,RAD51,RAD52, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 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。 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. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76600 |
DOI: | 10.6342/NTU201700600 |
全文授權: | 未授權 |
顯示於系所單位: | 食品科技研究所 |
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