大葉桃花心木 (Swietenia macrophylla) 在馬來西亞當地被認為是一種藥用植物，其種子在當地傳統醫學上常作為保健食品使用。在癌症領域上，較少有文獻使用大葉桃花心木進行癌症新藥的研究。因此本實驗想探討大葉桃花心木種子對於各種癌症細胞系的抑制作用，並從中篩選效果較好的細胞株進一步探討其作用機制。我們使用乙醇進行萃取大葉桃花心木種子 b 的成分並用來測試人類黑色素瘤細胞株 (A2058) 、人類乳癌細胞株 (MDA-MB-231,MCF-7) 、人類正常乳腺細胞株 (MCF-10A) 、人類肺癌細胞株 (A549) 以及小鼠胚胎成纖維母細胞 (NIH3T3) 透過 MTT assay 評估細胞毒性，並根據結果計算 IC50 ( Half maximal inhibitory concentration)，結果顯示大葉桃花心木種子乙醇萃取物對於 A2058 細胞株比起其他細胞株具有較好的毒殺能力，因此做為後續的研究對象。此外從Hoechst 螢光染色法觀察凋亡細胞，可以發現經由大葉桃花心木種子乙醇萃取物處理後的細胞產生出凋亡小體之現象，並且透過 DNA 片段化測試及 Annexin V/ 7-AAD 染色進一步證實大葉桃花心木種子乙醇萃取物確實用有誘導 A2058 細胞凋亡的能力。最後經由西方墨點法觀察凋亡相關蛋白表現量，結果顯示抗凋亡蛋白 Bcl-2 及存活蛋 Survivin 隨著大葉桃花心木種子乙醇萃取物濃度的升高，表現量均有明顯下降，並且 Caspase3 有被活化的跡象。在信號途徑結果顯示大葉桃花心木種子乙醇萃取物可能具有阻斷 MAPK 及 p-AKT 信號傳導的能力。總和上述結論，大葉桃花心木種子乙醇萃取物可能經由抑制 Survivin 與 Bcl-2 以及阻斷MAPK 及 p-AKT 信號傳導使 A2058 細胞走向細胞凋亡。

Swietenia macrophylla is a traditional medicinal plants. In Malaysia they are often used as Healthy food in traditional medicine. In the field of cancer, fewer use Swietenia macrophylla fruits as a research object. Therefore, this experiment wanted to investigate the inhibitory effect of Swietenia macrophylla fruits on various cancer cell lines, and to screen the cell strains with better anticancer effect to further explore its mechanism of action. We use the ethanolic extracts of Swietenia macrophylla seed to test the cytotoxicity in human breast cancer cells (MCF-7 and MDA-MB-231), human normal breast cells(MCF-10A), mouse fibroblast cells (NIH/3T3), human melanoma cells (A2058) and human lung cancer cells (A549). The cytotoxicity was assessed by conducting MTT assay. According to the experimental results, the half maximal inhibitory concentration (IC50 ) was calculated. The results showed that the Swietenia macrophylla seed extract had the highest inhibitory effects in A2058 cells than the other cancer cell lines, so it was used as a follow-up research object. Further, the apoptotic cells were observed by Hoechst assay and it was found that the cells produced apoptotic bodies after Swietenia macrophylla seed extracts treatment. The induction of apoptosis in A2058 cells following Swietenia macrophylla seed extracts treatment was further confirmed by DNA fragmentation and Annexin V / 7-AAD staining. The results by above apoptotic assays revealed that Swietenia macrophylla seed extract have the ability to induce apoptosis. Finally, the expression of apoptosis-related protein was observed by western blot assay. The results showed that Bcl-2 and Survivin showed significant decrease in expression with increasing drug concentration, and Caspase3 showed signs of activation. The results were by Western blot assay using phospho specific antibody indicated that Swietenia macrophylla seed extract may have the ability to block MAPK and p-AKT signaling. Based on the above conclusions, the Swietenia macrophylla seed extract may cause A2058 cells to undergo apoptosis by decreasing the expression of Survivin and Bcl-2 and blocking MAPK and p-AKT cell signaling.

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