臺灣博碩士論文加值系統

在多發性骨髓瘤(Multiple myeloma，簡稱 MM)細胞中，介白素6 (IL-6)所誘導的三個主要訊息傳遞路徑：PI3K / AKT、JAK / STAT、RAS / MAPK，會促進細胞的增生、抗凋亡及移動等現象。在MM細胞中，對Interferon-α(IFN-α)敏感的細胞株，IFN-α誘發的apoptosis是經由PML作用。而先前實驗室利用比較蛋白質體學鑑定出許多AKT可能的下游訊息分子，包括β-actin、cyclophilin A和promyelocytic leukemia protein (PML)等蛋白質。我們利用flow cytometer觀察AF-10細胞株在有無IFN-α處理之下，其apoptosis量的差異，確認AF-10是IFN-α敏感的細胞株。並利用 Western blot觀察PML在IL-6和wortmannin處理之下，其表現量的差異情形，確認 PML為 AKT substrate。將AF-10細胞以IFN-α和IL-6處理，有IL-6前處理的組別其apoptosis量減少。將細胞以cycloheximide處理然後觀察PML protein半衰期的變化，發現PML在IL-6處理下半衰期變短。

中文摘要…………………………………………………………………I
英文摘要………………………………………………………………...II
目錄……………………………………………………………………..III
一、背景簡介…………………………………………………………....1
二、研究目的……………………………………………………………7
三、材料與方法…………………………………………………………9
1細胞培養………………………………………………………….9
2免疫共沉澱……………………………………………………….9
3西方墨點法….…………………………………………………..10
4 Plasmid………………………………………..……………….11
5轉形作用…………………………………………………………11
6 Lentivirus製備………………………………………..……….12
7 Lentiviral transduction…………………………………...…….13
8 apoptosis………....……………………………………..……...13
四、實驗結果…………………………………………………………..15
1在AF-10細胞中PML是AKT的下游分子………………………..15
2 AF-10細胞為IFN-α敏感的細胞株…………………….…….16
3在IL-6刺激下，IFN-α引起的 apoptosis會減少………....16
4 AF-10細胞中IL-6減緩IFN-α引起的apoptosis不是經由
PI3K/AKT訊息傳導途徑….……………..……………………..17
5 AF-10細胞在IL-6處理之下PML protein半衰期的變化 ….18
五、討論………………………………………………………………...21
六、參考文獻……………………………………………………..……25
七、圖表……………………………………………………………..…27
八、附件………………………………………………………………..33

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