以活體噬菌體顯示法發展台灣地區口腔癌之標的治療

Using in Vivo Phage Display to Develop Targeted Therapy for Oral Ancer in Taiwan

鄭世緯(Shi-Wei Cheng)

指導教授 ： 郭彥彬

國立臺灣大學/醫學院/口腔生物科學研究所/碩士(2007年)

https://doi.org/10.6342/NTU.2007.01134

摘要

根據衛生署92 年癌症登記報告指出，口腔癌在臺灣男性十大癌症中標準化死亡率為第四位，年增率為第一位，影響國民健康甚劇。 口腔癌的治療包括手術治療合併放射線或是化學治療。 但這些治療方法多是非專一性的去除腫瘤，並未完全針對癌細胞的要害加以攻擊，都有嚴重的副作用， 且病人之五年存活率並未有明顯改變。 專一性的標的療法或許可以解決此一問題。
先前研究指出，利用活體噬菌體顯現法技術可以篩選出與腫瘤組織專一性結合的標的胜肽。本研究利用此方法對台灣口腔癌細胞株TW2.6異體移殖腫瘤組織進行五次的活體篩選， 找出可以和TW2.6異體移殖腫瘤組織結合之噬菌體株。 其中噬菌體株 TWO-5，可以和在異體移殖的TW2.6口腔癌腫瘤有專一性的結合。與口腔癌組織結合的能力分別高於其他對照器官(心、肺)約6到20倍。控制組的噬菌體株，則不具有與腫瘤組織專一性結合的能力。進一步利用合成的相對應胜肽 TWO-5P 與噬菌體株做活體競爭實驗，發現合成的胜肽可以抑制噬菌體株與腫瘤結合的能力。我們將合成的胜肽連結微脂體包覆抗癌藥物doxorubicin (TWO-5P-Lipo-Dox)，和單一只有此抗癌藥(Lipo-Dox)來進行抗癌療效比較。結果發現以TWO-5P-Lipo-Dox治療的SCID鼠效果最佳，腫瘤體積明顯小於以Lipo-Dox治療的小鼠。此外相較於Lipo-Dox治療的SCID小鼠，TWO-5P-Lipo-Dox治療的SCID鼠體重沒有明顯的減少，顯示TWO-5P-Lipo-Dox藥物所引起的副作用受到良好的控制。而我們在末次治療後，追蹤觀察小鼠之存活率，發現TWO-5P-Lipo-Dox(存活率100%)相較於其他兩組(Dox組存活率66.67%；PBS組存活率33.33%)，確實可以有效地提升存活率，因此TWO5P標的胜肽不但能提高腫瘤治療效果及專一性，也能減輕藥物的毒性,可提供未來一個口腔癌新穎的治療方式。

關鍵字

噬菌體顯示法 ； 口腔癌 ； 標的治療

並列摘要

According to the 2003 annual cancer registry report of department of health, oral cancer ranks fourth in the mortality rate of male cancer patient in Taiwan. The surgery combine cytotoxic chemotherapy or radiotherapy is the major treatment as cancer therapies. These therapies are not specific for malignant cells and the efficiency is limited by serious side effect. Therefore, despite significant improvements in diagnosis, local management, and chemotherapy of head and neck cancer, there is no significant increase in long-term survival rates over the past 30 years. New treatment strategies are needed to improve the survival rates for oral cancer.
Recent studies show that in vivo phage display is a powerful tool for the discovery of ligands that selectively home to tumor. Using this method, we identify several 12-mer peptides specifically binding to xenografts of a Taiwanese oral cancer cell line TW2.6. One selected phage clone, TWO-5 specifically bound to the TW2.6 xenograft and the effects was inhibited by competition with phage-derived peptides. Furthermore, TWO-5-peptide-linked liposomes that carried doxorubicin (TWO-5P-Lipo-Dox) not only suppressed tumor growth better than Lipo-Dox but also showed no significant body weight change. TWO- 5P-Lipo-Dox treated mice have 100% survival rate but Lipo-Dox and PBS treated mice only have 66.67% and 33.33% survival rate. These results indicate that TWO-5 peptide enhanced the efficacy of the drug against oral cancer xenografts in SCID mice and suggest that this peptide has a strong clinical potential for drug delivery guider to treat oral cancer.

並列關鍵字

phage display ； oral cancer ； targeted therapy

參考文獻

Arap, W., M. G. Kolonin, et al. (2002). "Steps toward mapping the human vasculature by phage display." Nat Med 8(2): 121-7.

Arap, W., R. Pasqualini, et al. (1998). "Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model." Science 279(5349): 377-80.

Augustin, H. G., D. H. Kozian, et al. (1994). "Differentiation of endothelial cells: analysis of the constitutive and activated endothelial cell phenotypes." Bioessays 16(12): 901-6.

Borsum, T., I. Hagen, et al. (1982). "Alterations in the protein composition and surface structure of human endothelial cells during growth in primary culture." Atherosclerosis 44(3): 367-78.

Chen, Y. K., H. C. Huang, et al. (1999). "Primary oral squamous cell carcinoma: an analysis of 703 cases in southern Taiwan." Oral Oncol 35(2): 173-9.