桑椹花青素誘導WEHI-3血癌細胞凋亡及分化之研究

The Study of Mulberry Anthocyanins Inducing Leukemia WEHI-3 Cells Apoptosis and Differentiation

邵思雅(Sz-Ya Shao)

指導教授 ： 黃惠珮

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2012年)

摘要

花青素 (anthocyanins) 廣泛存在植物界中在帶有紅、藍、紫顏色的水果和蔬菜中，桑椹也含豐富的花青素。對於人體的健康，高花青素濃度的飲食攝取是非常重要的。本研究室也證實桑椹之花青素可抑制多種癌細胞產生凋亡，如：胃癌細胞、黑色素細胞癌、肝癌等。目前對於血癌的治療效果有限，多半採用化學療法或給予抗癌藥物，往往會使病人產生嚴重的副作用，而這些藥物對正常細胞也會產生影響。因此，現在許多研究趨向從天然萃取物中尋有效成分來做為治療的參考或做為複方，由於目前沒有相關研究證明桑椹花青素能使小鼠血癌細胞株 (WEHI-3) 產生凋亡，本實驗將探討桑椹花青素對小鼠血癌細胞株 (WEHI-3) 和WEHI-3誘導之血癌老鼠的影響。結果指出，桑椹花青素對WEHI-3細胞有毒殺作用，由流式細胞儀分析結果發現，桑椹花青素能讓細胞週期停滯於G0/G1，最後走向凋亡，從DAPI染色、粒線體膜電位檢測及Annexin-v/PI雙染實驗中得到證實。在PMA誘導WEHI-3細胞分化的實驗中，桑椹花青素具有促進分化的能力，並觀察到有高GM-CSF生長因子的表現、表面抗原CD14，CD11b、吞噬能力的增加。在動物實驗中，高濃度桑椹花青素處理的組別可以延緩血癌老鼠的發病進程，增加其存活率，減少不成熟白血球細胞出現在血液中、減少肝臟和脾臟中白血病細胞之浸潤。綜合以上的結果顯示，桑椹花青素可促使癌細胞凋亡及分化延緩血癌老鼠發病的進程，期許其可用來做為使用化學預防物質的參考。

關鍵字

血癌 ； 桑椹花青素

並列摘要

Anthocyanins exist widely in many bright red, blue and purple colors of vegetables and fruits, include mulberry. It potential importance to human health is the relatively high concentration of anthocyanins in the diet. In our previous studies, we showed that Mulberry anthocyanins (MACs) have the antitumor effects on various cancers, such as gastric cancer, melanoma, and hepatoma. However, cancer therapy is limited, acute myeloid leukemia (AML) patients usually suffer from the side effects of chemotherapy, and affect normal cell. Recently, interest in dietary phytochemicals and Chinese herbs for potential cancer chemoprevention has increased substantially owing to their multiple capacities of a combination. There is no available information addressing MACs-affected apoptotic responses in the murine myelomonocytic leukemia cell line (WEHI-3). We investigated that MACs induced cytotoxic effects in vitro and affected WEHI-3 cells in vivo. This study showed WEHI-3 cells numbers were decreased after treatment with MACs by MTT assay. The results of flow cytometry assay showed that MACs caused an apparent accumulation of cells in the G0/G1 phase and chromatin condensation, reduced the level of ΔΨm and increased apoptosis cell level by Annexin-v/PI experiment. In addition, MACs could promoted differentiation in WEHI-3 cells, increased the level of GM-CSF, CD14, CD11b and phagocytic ability. Besides, MACs enhanced the survival rate of AML mice, reduced tumor cell infiltrate liver and spleen, decreased blast cell number in blood in vivo. In conclusion, MACs can prevent the growth of WEHI-3 cells for potentially combination chemotherapy therapy to prevent side effects of acute myeloid leukemia.

並列關鍵字

leukemia ； Mulberry Anthocyanins

參考文獻

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