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National Chung Hsing University Institutional Repository - NCHUIR > 生命科學院 > 生物化學研究所 > 依資料類型分類 > 碩博士論文 >  蘋婆乙酸乙酯精萃物可過度表現凋亡誘導因子致三陰性乳癌細胞凋亡

Please use this identifier to cite or link to this item: http://nchuir.lib.nchu.edu.tw/handle/309270000/152336

標題: 蘋婆乙酸乙酯精萃物可過度表現凋亡誘導因子致三陰性乳癌細胞凋亡
Ethyl Acetate Extract of Sterculia nobilis Induces Apoptosis of Triple Negative Breast Cancer Cells Through the Overexpression of Apoptosis Inducing Factor
作者: 張恩慈
Chang, En-Tzu
Contributors: 高振益
Jung-Yie Kao
生物化學研究所
關鍵字: 蘋婆;凋亡誘導因子;三陰性乳癌;細胞凋亡
Sterculia nobilis;apoptosis inducing factor;triple negative breast cancer;apoptosis
日期: 2013
Issue Date: 2013-11-07 13:21:17 (UTC+8)
Publisher: 生物化學研究所
摘要: 本篇研究利用蘋婆果莢乙酸乙酯精萃物(Ethyl acetate extract shell of Pod Sterculia Nobililis,簡稱為EPSN)針對人類三陰性乳癌細胞株BT-20與MDA-MB-231進行細胞毒殺實驗。藉由細胞存活率測試(MTT assay),經EPSN處理48小時的IC50,BT-20為40-50 μg/mL,和前人研究結果相類似(30-40 μg/mL)。由前人研究得知BT-20經EPSN處理後可能是藉由細胞凋亡誘導因子(Apoptosis-inducing factor,AIF)經凋亡蛋白酶非依賴型路徑(caspase-independent pathway)來調控細胞凋亡。
本篇研究調整EPSN處理細胞的時間並於收取細胞蛋白時分離細胞質與粒線體兩部分,經西方墨點法分析發現BT-20與MDA-MB-231皆於加入EPSN 2小時後,AIF在細胞質中有過度表現(overexpression)的情形,進一步利用Real-time PCR觀察BT-20經EPSN處理3小時後的AIF mRNA表現量,發現AIF mRNA有明顯上升趨勢,由此可得知蘋婆果莢乙酸乙酯精萃物在短時間內即具有誘導三陰性乳癌細胞凋亡的能力。
In previous studies at our laboratory, we found that the Ethyl acetate extract, isolated from shell of pod of Sterculia nobililis (EPSN), can induce the apoptosis of human triple-negative breast cancer cell line BT-20 through the regulation of apoptosis-inducing factor (AIF) in caspase-independent pathway. In this study, we used EPSN to treat BT-20 cells for cytotoxicity test (48 Hr IC50:40-50 μg/mL) and to investigate how does AIF regulate the mechanism of apoptosis in cancer cells.
The results of Western blotting analysis revealed that, in both BT-20 and MDA-MB-231 cell lines, AIF was started to overexpress in cytosol compartment after 2 hours treatment of EPSN. We further used Real-time PCR to observe the mRNA expression level AIF in BT-20 after the treatment of EPSN. The data showed that there was indeed a signicant increment expressed in mRNA level of AIF after 3 hours treatment of EPSN. These results demonstrated that EPSN can induce apoptosis at the early stage in triple-negative breast cancer cells with the mRNA and protein overexpressions of AIF leading to the cell death through the caspase-independent pathway.
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