探討抗菌胜肽m2163與人類大腸癌細胞SW480之間的作用機制

Interaction mechanism between antimicrobial peptide, m2163 and human colon adenocarcinoma, SW480

陳怡潔(Yi-Chieh Chen)

指導教授 ： 林志侯

國立清華大學/生命科學院/分子醫學研究所/碩士(2013年)

摘要

m2163是一帶正電抗菌胜肽，由實驗室發表文獻已知其可抑制數種乳酸菌及李斯特菌的生長，且對大腸癌細胞SW480也有生長抑制及細胞凋亡的現象，但其造成SW480生長受抑制與細胞凋亡的原因尚不清楚，本實驗希望能找出m2163與SW480之間的作用機制，透過螢光標定m2163及流式細胞儀的偵測，證實m2163可與SW480結合，經螢光顯微鏡和共軛焦顯微鏡觀察到m2163乃作用於SW480細胞膜，由於文獻指出部分造成癌細胞細胞凋亡的抗菌胜肽是作用在粒線體，於是進一步將粒線體純化，但並沒有在粒線體偵測到m2163，推測粒線體並非m2163的作用位置，此外也針對細胞膜通透性加以分析，發現m2163會破壞細胞膜完整性，且發生在細胞凋亡之前，推論m2163透過作用於細胞膜並破壞細胞膜完整性，後續造成細胞生長受到抑制以及誘發細胞凋亡。

關鍵字

抗菌胜肽 ； HASH(0x81d9570) ； 大腸癌 ； 細胞凋亡

並列摘要

m2163 is a cationic antimicrobial peptide that inhibits SW480 cells proliferation and induces apoptosis. However, the process by which m2163 kills SW480 cells is not well understood. This study shows that apoptosis and proliferation inhibition in SW480 cells were precedes by m2163 binding to, and progressive permeabilization of the cell membrane. Flow cytometry reveals that m2163 interacts with SW480 cells. In addition, fluorescent microscopy and confocal microscopy show that m2163 localizes with cell membrane in SW480 cells. Mitochondria may not the target of m2163 because purified mitochondria which isolated after treated with FITC-m2163 were not detected fluorescent signal by flow cytometry. Furthermore, m2163 disrupts SW480 cell membrane integrity prior to apoptosis induction. These data suggest that interaction mechanism between m2163 and SW480 cells results from localization with and permeabilization of the cell membrane.

並列關鍵字

confocal ； locolization ； permeabilization

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