人類大腸直腸癌是一個非常嚴重的腫瘤疾病，而目前醫學上，對於大腸直腸癌的化學療法效果上卻很有限。而在台灣民間，由傳統中草藥M. australis (Moraceae)所分離出的粗萃取物被廣泛用在治療腫瘤疾病上，不過其確實的治療機制卻不甚明暸。Morusin是由M. australis (Moraceae)的根部中分離出的成份，具有引發癌細胞進行細胞凋亡的效果。在以morusin處理HT-29細胞6天後，其50%抑制濃度(IC50)為5.95 M，而以morusin處理PBMC細胞，其IC50為 29.75 M。在以morusin做軟洋菜膠分析14天後，其IC50為 2.76 M。在以morusn處理HT-29細胞後，可以發現細胞膜內側的磷脂絲胺酸有翻轉至外側細胞膜，細胞的sub-G1期的DNA有增加，有DNA片段化現象出現，並造成細胞核濃縮及出現凋亡小體現象，由以上結果證實morusin經由誘發細胞凋亡而抑制HT-29細胞生長。由西方墨點法分析morusin誘發HT-29細胞進行細胞凋亡的途徑，發現會造成caspase-8活化，並造成Bax、Bak活化，並降低Bcl-XL表現，並使用rhodamine 123染色，發現粒線體膜電位喪失，並造成cytochrome c及Smac/DIABLO由粒線體中釋放到細胞質中，另外抑制XIAP表現，接著活化caspase-9、caspase-3，最後造成細胞核中的PARP切割，以及使DFF45/ICAD被切割失去活性，造成細胞核內DFF40/CAD活化，最終造成細胞DNA斷裂，造成細胞凋亡現象出現。本研究證實morusin能經由活化caspase並影響粒線體的途徑造成大腸直腸癌細胞凋亡，本研究的結果，在未來可提供為抗癌藥物研發之參考。

　　Colorectal cancer is one of the leading causes of malignant death in western countries as well as in Taiwan. The crude extracts of M. australis (Moraceae) are common remedy to treat human colorectal cancer in Taiwan. However, the action mechanism of this crude extract is still unclear. The specific goal of the present study is to explore the anti-tumor mechanism of morusin, isolated from of M. australis, in human colorectal cancer, and to better understands if it can be used as a chemotherapeutic agent for cancer. The present study demonstrated that the IC50 of morusin on the growth of human colorectal cancer HT-29 cells was 5.95 M as compared to peripheral blood mononuclear cells was 29.75 M at day 6 of post-treatment. The inhibitory efficacy of morusin on colony formation of HT-29 in soft agar was 2.76 M at day 14 of post-treatment. The susceptibility of cells to morusin is caused by apoptosis through phosphtidylserine exposure, increase of sub-G1 content, DNA fragmentation, and nuclear morphologic change. Morusin induced caspase-8 activation, loss of mitochondria membrane potential, releases of cytochrome c and Smac/DIABLO from mitochondria to cytosol, decreased expression of XIAP, further activated caspase-9 and the caspase-3, which in turn cleaved DFF45/ICAD and PARP, and increased nuclear PARP and DFF40/CAD and DNA fragmentation. Our findings demonstrated that morusin induced HT-29 cell death via apoptosis. The molecular mechanism of morusin may provide new ideal for anti-cancer drug design.

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校內：2005-08-25公開
校外：2009-08-25公開