香茅精油和檸檬醛對人類惡性黑色素瘤A375 細胞株生長抑制與誘導細胞凋亡之分子機制探討

Molecular mechanism of lemongrass oil and citral induced growth inhibition and apoptosis in human malignant melanoma A375 cell lines

周子妤(Tzu-Yu Chou)

指導教授 ： 張基隆

高雄醫學大學/醫學院/醫學系生物化學科碩士班/碩士(2016年)

摘要

香茅(Lemongrass)是一種被廣泛作為食品香料草藥食物，並有止痛和消炎功效，其主成分檸檬醛(Citral)被證實具誘導細胞凋亡動能。本研究在探討香茅精油(Lemongrass oil)及檸檬醛對抑制人類惡性黑色素瘤細胞株A375細胞生長的抗癌機制及細胞凋亡路徑。利用香茅精油及檸檬醛作用A375細胞後分析對細胞存活率影響。結果顯示隨著香茅精油及檸檬醛濃度增加和作用時間延長，呈劑量及時間比抑制A375細胞存活。以流式細胞儀偵測活性氧化物(Reactive oxygen species, ROS)和細胞週期進行(Cell cycle progression)及去氧核醣核酸片段分析與TUNEL檢測DNA受損，顯示香茅精油及檸檬醛30~50 μg/ml會誘導A375細胞活性氧化物生成、G2/M細胞週期停滯及凋亡(Apoptosis)。透過西方墨點法得知香茅精油能減少Cdc25c和Cyclin B1蛋白表現使週期停滯在G2/M時期，以及降低細胞凋亡內在路徑之抗凋亡蛋白Bcl-2和Bcl-XL及增加凋亡蛋白Bax的表現，進而活化Caspase-9和Caspase-3等下游訊息路徑。以及透過外在路徑Fas Ligand與接受體(Death receptor) Fas receptor結合傳遞訊息，啟動下游訊息路徑，共同活化下游Caspase-3誘導細胞凋亡死亡。綜合以上結果顯示香茅精油及檸檬醛或許具有預防及治療人類惡性黑色素瘤之潛能。

關鍵字

香茅 ； 香茅精油 ； 檸檬醛 ； 黑色素細胞癌

並列摘要

Lemongrass one kind of herb, is widely used as a flavoring of food, and is used to lessen pains by it’s analgesic and anti-inflammatory properties. Previous research indicated citral is the major effective compound of lemongrass and has the potential to induce apoptosis of cells. The present study is to explore the effects of lemongrass oil and citral on human malignant melanoma cell line (A375) and explore the underline mechanism. Results showed that lemongrass oil and citral inhibited A375 cells growth in both time-dependent and dose-dependent manner. At the concentrations ranged from 30μg/ml to 50 μg/ml, either lemongrass oil or citral could stimulate production of reactive oxygen species (ROS), arrest cell cycle progress in G2/M phase by decreasing the synthesis of cyclin B1 and cdc25c, and induce apoptosis of human malignant melanoma A375 cells. Moreover, the expressions of antiapoptotic protein of Bcl-2 and Bcl-XL were down-regulated, whereas expressions of proapoptotic protein Bax, caspase-9, and caspase-3 were up-regulated.Moreover, Fas ligand and receptor expressions were also up-regulated which could activate caspase-3. Changes of those leaded to occurence of apoptosis.These results suggest that lemongrass oil and citral may have the potential in prevention against human malignant melanoma. However, further studies are needed to accumulate enough evidences to support this viewpoint.

並列關鍵字

Lemongrass ； Lemongrass oil ； Citral ； melanoma

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