修饰化爱玉果胶及其水解产物对人类胃癌细胞AGS之影响.DOC

修飾化愛玉果膠及其水解產物對人類胃癌細胞 (AGS) 之影響 The effect of modified jelly fig pectin and its hydrolysates on human stomach cancer cells (AGS) 研究生：潘思妤 Pan, Szu-Yu 指導教授：廖遠東 Liaw, Ean-Tun 【摘要】 近年來，柑橘果膠逐漸成為癌症研究者感興趣之營養成分，而柑橘果膠經pH的調整而成為短鏈、不分支且富含半乳糖的直鏈，其可有效的被人體吸收與利用。先前的研究指出，經修飾之柑橘果膠對於各類腫瘤細胞均有顯著的抑制效果，研究亦指出果膠可促使癌細胞走向凋亡。因此，本研究之目的在於探討經修飾之愛玉果膠對癌細胞是否亦具有抑制之效果，藉以提升其附加價值。目前以MTT方法已篩選出六種不同聚合度之果膠樣品，其對胃癌細胞具有較佳之抑制效果，其對細胞作用至72小時之抑制率可達50% 以上。而以倒立式位相差顯微鏡觀察其細胞形態，亦發現細胞凋亡之特徵，進而以流式細胞儀偵測其細胞週期、早期凋亡以及caspase-3活化之情形。在細胞週期方面，細胞經處理72小時後，其sub-G1之比例介於22-36%。在早期凋亡方面，細胞經處理48小時後，其早期凋亡之比例達到最高，比例介於11-30%。而在caspase-3活性測定方面，亦有發現活化之情形。由上述結果可初步證實，愛玉果膠經鹼和酵素適度修飾後，確實能使胃癌細胞走向凋亡。 關鍵字：修飾化柑橘果膠、細胞凋亡、愛玉、流式細胞儀 【Abstract】 A nutrient of recent interest to cancer researchers is citrus pectin. However, citrus pectin is modified (MCP) by adjusting the pH that breaks it into shorter, non-branched, galactose-rich chain that are better absorbed and utilized by the body. Previous studies have shown that the modified citrus pectin could definitely inhibit lots of tumor cells. Furthermore, the studies have indicated that pectin also can induce cancer cells undergoing apoptosis. Therefore, the aim of present study is to investigate the cancer cell inhibition by modified jelly fig pectin and its hydrolysates for promoting its added-values. By MTT assay, six hydrolyzed samples were prepared and they showed effective inhibition on AGS stomach cancer cells. The inhibition rate can be achieved up to 50% when the tumor cell treated with these samples for 72 hours. By Reverse Phase microscope, cell apoptosis morphology was observed. Simultaneously, cell cycle analysis, early apoptosis, and caspase-3 activation were conducted by flow cytometry. In cell cycle, the results indicated that the sub-G1% could reach 22-36% after treated with all of the samples for 72 hours. In early apoptosis investigation, the data suggested that 11-30% early apoptosis can be found when hydrolyzed samples were employed for 48 hours. Moreover, in caspase-3 activity assay dem