質譜分析化療藥物對粒線體磷脂質成分的影響

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Please use this identifier to cite or link to this item: http://140.128.103.80:8080/handle/310901/31353

Title:	質譜分析化療藥物對粒線體磷脂質成分的影響
Other Titles:	Chemotherapeutic Drug Effects on Glycerophospholipid Composition of Mitochondria Analyzed by Mass Spectrometry
Authors:	賴若蕓 LAI,RUO-YUN
Contributors:	許員豪 Hsu, Yuan-Hao 化學系
Keywords:	磷脂醯甘油;磷脂醯膽鹼;腦磷脂;磷酸絲氨酸;心磷脂;質譜技術 Phosphatidylglycerol;Phosphatidylcholine;Phosphatidylethanolamine;Phosphatidylserine;Cardiolipin;Mass Spectrometry
Date:	2019
Issue Date:	2019-03-21T08:59:59Z (UTC)
Abstract:	粒線體膜脂質的成分有心磷脂、磷脂醯膽鹼、腦磷脂等，引發我想探討化療藥物對癌細胞粒線體脂質組成的改變，因此先建立質譜偵測甘油脂質的平台，由LC-MS分析每種脂質，再進行生物樣品的偵測，我們能夠鑑定出五種磷脂，包含磷脂醯甘油、磷脂醯膽鹼、腦磷脂、磷酸絲氨酸以及心磷脂。我們以Hep G2為控制組，以添加50 μM 5-fluorouracil當作實驗組，來探討化療藥物對肝癌細胞的影響。我們接著以RT-PCR的實驗觀察粒線體相關基因變化，發現加藥之後CRLS1有明顯下降。更進一步以質譜分析心磷脂的種類變化，發現50 μM 5-fluorouracil 加藥培養24小時，相較於控制組在心磷脂長碳鏈的部分，飽和度有增加，培養48小時飽和度也顯著的增加。接續趙昱任與賴冠霖的實驗偵測大部分的磷脂質，HT-1080隨著Mitomycin C與Methotrexate的作用時間增加，腦磷脂、心磷脂的含量皆有增加的趨勢，其飽和度也增加。 The components of mitochondrial membrane lipids include many phospholipids,　such as cardiolipin, phosphatidylcholine,　phosphatidylethanolamine, phosphatidylglycerol and phosphatidylserine. Therefore, we added chemotherapeutic drugs to observe the lipid composition changes of the mitochondria in cancer cells. We first established a platform for the detection of lipids by mass spectrometer, and each lipid was analyzed by LC-MS. We then applied this quantification method to detect the phospholipids in the biological samples. Up to date, we are able to identify five groups of phospholipids, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and cardiolipin. We used Hep G2 as a control group and 50 μM 5-fluorouracil as an experimental group to investigate the effect of chemotherapeutic drugs on hepatoma cells. We used RT-PCR experiments to observe mitochondrion-related gene changes and found that CRLS1 was significantly decreased after dosing. Furthermore, we analyzed the change in cardiolipin by mass spectrometry. We found that 50 μM 5-fluorouracil was cultured for 24 hours compared with the control group, the degree of saturation was increased in the long chain of cardiolipin, and the saturation was significantly increased in culture for 48 hours. Following the experiment of Chao Yu-Jen and Lai Guan-Lin, most of the phospholipids were detected. The interaction time of HT-1080 with Mitomycin C and Methotrexate increased, phosphatidylethanolamine and cardiolipin increased, and the saturation increased.
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