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研究生:蔡宗憲
研究生(外文):Tsung Hsien Tsai
論文名稱:探討口型蛋白如何藉由控制表皮生長因子受體所調節的細胞外調節蛋白激酶來調控脂肪新生
論文名稱(外文):Stomatin mediates the adipogenesis via regulating the EGFR stimulated ERK pathway
指導教授:林奇宏林奇宏引用關係
指導教授(外文):Chi Hung Lin
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:微生物及免疫學研究所
學門:生命科學學門
學類:微生物學類
論文種類:學術論文
論文出版年:2020
畢業學年度:108
語文別:中文
論文頁數:68
中文關鍵詞:口型蛋白質脂肪細胞脂肪新生細胞外調節蛋白激酶表皮生長因子接受器
外文關鍵詞:StomatinAdipocyteAdipogenesisERKEGFR
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  • 被引用被引用:0
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目錄.....................................................i
摘要....................................................iii
Abstract.................................................iv
1.緒論....................................................1
1.1口型蛋白質(Stomatin)....................................1
1.2口型蛋白的分布與功能.....................................1
1.3脂肪組織與脂肪新生(Adipogenesis)........................2
1.4細胞外調節蛋白激酶(extracellular signal‑regulated kinases ,ERK).....................................................3
1.5表皮生長因子接受器(EGFR) 與表皮生長因子接受器內化作用(EGFR internalization)..........................................4
2.實驗材料與方法...........................................6
2.1細胞培養與3T3-L1脂肪前驅細胞分化.......................6
2.2架構質體(plasmid)和細胞轉染(transfection).............6
2.3慢病毒(Lentivirus)系統與感染3T3-L1 脂肪前驅細胞........7
2.4共軛交顯微鏡螢光影像..................................8
2.5西方墨點與分析........................................9
2.6油紅O (Oil Red O)染色與定量油滴累積方法................9
2.7生長因子接受器(EGFR)降解實驗............................10
2.8 MAPK pathway 抑制劑刺激實驗...........................11
2.9 Reverse transcription PCR (RT-PCR) 與 Real-time PCR (Quantitative PCR,qPCR ).................................11
2.10統計分析..............................................12
3.結果...................................................13
3.1降低口型蛋白質表達可增加脂肪分化後期ERK的活化作用.........13
3.2口型蛋白降低所調控的ERK的活化可能具有調控分化之能力.......13
3.3口型蛋白與表皮生長因子接受器之相關性.....................14
3.4口型蛋白無法調控表皮生長因子接受器之降解作用..............15
3.5口型蛋白調控表皮生長因子接受器之內化作用(Internalization)......................................16
4. 討論..................................................20
4.1探討口型蛋白質敲弱組別ERK活化上升與MAPK路徑抑制劑實驗結果...20
4.2探討口型蛋白質與表皮生長因子接受器的關聯性................21
5.參考文獻................................................24
6.圖.....................................................26
7.附錄...................................................63
附錄表格一...............................................63
附錄表格二...............................................64
附錄圖表.................................................65
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16 Umlauf, E. et al. Association of stomatin with lipid bodies. Journal of Biological Chemistry 279, 23699-23709 (2004).
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21 Madshus, I. H. & Stang, E. Internalization and intracellular sorting of the EGF receptor: a model for understanding the mechanisms of receptor trafficking. Journal of cell science 122, 3433-3439 (2009).
22 Kim, S. et al. Graf regulates hematopoiesis through GEEC endocytosis of EGFR. Development 144, 4159-4172 (2017).
23 Yamamoto, N., Yamashita, Y., Yoshioka, Y., Nishiumi, S. & Ashida, H. Rapid preparation of a plasma membrane fraction: Western blot detection of translocated glucose transporter 4 from plasma membrane of muscle and adipose cells and tissues. Current Protocols in Protein Science 85, 29.18. 21-29.18. 12 (2016).
24 Pagano, E. & Calvo, J. C. ErbB2 and EGFR are downmodulated during the differentiation of 3T3‐L1 preadipocytes. Journal of cellular biochemistry 90, 561-572 (2003).
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27 Chen, C.-Y. et al. Decreased expression of stomatin predicts poor prognosis in HER2-positive breast cancer. BMC cancer 16, 1-8 (2016).
28 An, H. et al. Stomatin plays a suppressor role in non-small cell lung cancer metastasis. Chinese Journal of Cancer Research 31, 930 (2019).
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