The synergistic anticancer effect of Lipo-Dox and perforin on colon cancer cell, CT26

Abstract

[[abstract]]穿孔素是一種經由活化態細胞毒殺型T淋巴球及自然殺手細胞所表現的分泌型蛋白質。穿孔素在毒殺細胞的機制中為一重要的物質，其作用為插入目標細胞之細胞膜並在細胞表面形成孔洞，造成細胞傷害。在先前的研究中，我們探討利用穿孔素配合抗癌藥物Doxorubicin的協同效用對於毒殺癌細胞的可能性。穿孔素表現載體經由CMV啟動子所驅動，並將表現載體轉染至小鼠大腸癌細胞株，CT26。透過MTT分析的方式評估不同表現載體穿孔素毒殺效率。構築分別設計接有小鼠內皮細胞生長因子之訊號序列及去除特定C端序列的穿孔素，使其對癌細胞的毒殺效果增強。由實驗結果可知，表現載體所表現的穿孔素對癌細胞具有細胞毒性。此外，接上血管內皮生長因子之訊號序列及去除C端的穿孔素表現載體，對癌細胞的毒殺能力皆有增強效果。本研究成果提供一具潛力的抗癌治療平台；同時，本治療策略可減少藥物用量，因此能減少抗癌藥物對患者所產生的副作用。[[abstract]]Perforin, a secreted protein, is synthesized by activated cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. It is a key component of the lytic machinery, being able to insert into the plasma membrane of targeted cells to form pores which lead to cell destruction. In the present study, we investigated the possibility of tumor eradication through the effect of perforin and its synergistic effect with antitumor drug, doxorubicin (Dox). Several perforin plasmids, driven by CMV promoter, were constructed and were transfected to murine colon cancer cell line, CT26. The cytotoxicity of perforin was evaluated by MTT assay. The flanking of mouse VEGF signal sequence and the deletion of C-terminus sequence on perforin at DNA level were performed to enhance the cytotoxicity against cancer cells. This synergistic effect of perforin and Dox suggested a potential strategy for cancer therapy and to reduce antitumor drugs caused side effects.[[note]]碩