山胡椒对人类乳癌细胞株MCF.doc

山胡椒對人類乳癌細胞株MCF-7細胞毒性之探討 Evaluation of cytotoxicity of Litsea cubeba Pers. extracts in human breast carcinoma cell line MCF-7 研究生：許惠茹 Huei-Ru Shiu 指導教授：許祥純 Shyang-Chwen Sheu 根據行政院衛生署統計，97年國人十大死因之首為惡性腫瘤，其中乳癌為婦女癌症的第二位，研究指出許多植物具有防治癌症之功效，因此由植物中篩選出天然的抗癌成分，為一重要之研究課題。本研究以 Litsea cubeba Lour. Persoon 各部位之甲醇及乙醇萃取物，評估其對於人類乳癌細胞株MCF-7之細胞毒性，實驗以5-Fluorouracil作為正控制組，並以山胡椒各部位的甲醇及乙醇萃取物處理人類乳癌細胞株MCF-7 24小時，結果顯示山胡椒果實、樹葉、枝條及花苞甲醇萃取物IC50值分別為49.47 ± 1.09、191.09 ± 20.45、60.36 ± 7.3881.20 ± 5.47 μg/mL，而乙醇萃取物之IC50值則依序為32.32 ± 0.86、188.98 ± 16.60、103.36 ± 12.49及103.70 ± 1.45 μg/mL，顯示果實乙醇萃取物對人類乳癌細胞株MCF-7的毒殺效果最佳；以位相差顯微鏡觀察細胞死亡型態，山胡椒萃取物處理於人類乳癌細胞株MCF-7之細胞死亡型態為細胞皺縮並且使細胞密度下降及形成群聚的圓形細胞，實驗進一步萃取細胞DNA發現，DNA並未產生斷裂或拖曳現象，再以流式細胞儀評估人類乳癌細胞株MCF-7細胞週期之變化，結果發現山胡椒萃取物會使細胞週期停滯於S期，以西方墨點法評估細胞週期調控分子之蛋白質表現性，顯示山胡椒果實、花苞甲醇萃取物以及果實、枝條乙醇萃取物會誘導細胞週期調控分子Cyclin A、CDK2及E2F-1表現量增加，綜合實驗結果得知山胡椒萃取物可藉由調控人類乳癌細胞株MCF-7之細胞週期調控分子表現，而促使細胞週期停滯，誘導細胞凋亡 關鍵字：乳癌細胞株MCF7、抗腫瘤、細胞凋亡、細胞週期停滯 Abstract】 From the statistics announced by Department of Health of Taiwan, cancer is the first of top 10 causes of death. Breast cancer is the second highest of women’s cancer. Many reports have indicated that plants have anti-cancer effect. Therefore, isolating effective antitumor compounds from plants is an important task. In this study, the cytotoxicity of methanol and ethanol extracts of Litsea cubeba Lour. Persoon in human breast carcinoma cell line MCF-7 was evaluated. 5-Fluorouracil was used as a positive control. Results showed that the IC50 values of methanol extracts of fruits, leaves, branches and flower buds from L. cubeba were 49.47 ± 1.09, 191.09 ± 20.45, 60.36 ± 7.38 and 81.20 ± 5.47 μg/mL, respectively. The IC50 values of ethanol extracts were 32.32 ± 0.86, 188.98 ± 16.60, 103.36 ± 12.49 and 103.70 ± 1.45 μg/mL, respectively. The ethanol extract of fruits showed the highest cytotoxicity. The morphology of MCF-7 cells treated with L. cubeba extracts was shrinkaged, decreased cell density and formed cluster of round cells. Furthermore, DNA extracte