臺灣博碩士論文加值系統

Photofrin® 是第一個用於光動力療法 (photodynamic therapy, PDT) 的光顯劑 (photosensitizer), 可用於治療癌症。本實驗室研究以Photofrin®進行的光動力療法過程中發現Photofrin®可以分佈在癌細胞內部不同的位置，而隨著分佈的位置不同，PDT所引發的細胞死亡型態與機制亦不相同。在研究的過程中發現到一個有趣的現象，不論照光與否，以Photofrin®處理人類細胞會產生可被anti-caspase-3抗體所辨識的高分子量的蛋白質。此高分子量的蛋白質會隨著Photofrin®處理的時間與劑量增加而增加；除了內生性的caspase-3，外送過量表現的caspase-3也可觀察到相同的現象。我們的實驗結果指出此高分子量的蛋白質是來自已存在細胞中的caspase-3經由共價鍵結的修飾作用所形成，而且極可能是兩個或三個caspase-3單體分子鍵結在一起。Photofrin®的前處理會抑制由抗癌藥物所引起之細胞凋亡的訊息傳導，例如caspase-3的活化、PARP的斷裂及PS的外翻，但細胞的存活度卻反而下降。我們推測這樣的修飾作用在細胞凋亡的過程中可能會影響caspase-3的活化，進而影響細胞死亡的機制。Caspase-3在細胞凋亡的訊息傳遞過程中扮演著極為重要的角色，而此種造成caspase-3的修飾作用尚未被報導過。基於我們的實驗結果認為此種caspase-3的修飾作用在細胞死亡的訊息傳遞過程中可能極為重要。

Photofrin® is the first generation photosensitizer for photodynamic therapy (PDT), which is approved for treatment of cancer and some other diseases. Previous work from our laboratory indicated that PDT with Photofrin® targeting to distinct subcellular site results in different death phenotypes of A431 cells (Hsieh et al., 2003). Following that study, we found that Photofrin® induced formation of the two high-molecular- weight (HMW) forms of caspase-3 in various human cell lines. Photofrin® induces the formation of two HMW forms of caspase-3 is dose- and time-dependent, and this modification could also be observed on ectopically expressed caspase-3. Our results showed that the Photofrin®—induced HMW forms of caspase-3 were resulted from dimerization or trimerization of caspase-3 monomer through covalent conjugation. Pretreatment of Photofrin® inhibited the apoptotic processes triggered by staurosporine and anti-cancer drugs taxol and etoposide, including the cleavage/activation of caspase-3, the cleavage of PARP, and PS externalization, suggesting that the modification of caspase-3 might affect the activity of caspase-3 in apoptotic cells. Because caspase-3 is the key executioners in apoptosis, and our findings reveal a novel mechanism for regulating caspase-3 functions in cells.

指導教授推薦書…………………………………………………………
口試委員會審定書………………………………………………………
授權書………………………………………………………...…………iii
誌謝……………………………………………………………...………iv
中文摘要………………………………………………………...……….v
Abstract…………………………………………………….…...………vi
Content……………………………………………………….………….vii
Introduction……………………………………………...……………….1
Meterials and methods……………………………………………………6
Results…………………………………………………………………...14
Discussions……………………………………………………………...23
References……………………………………………………………….27
Figures and Table………………………………………………………..31

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