中草藥是一個主要天然的資源，在癌症的治療上已被廣為人所知。它的優點包括低副作用、血管新生及轉移的抑制作用。肝細胞癌，在東南亞是一個常見的人類惡性腫瘤，它們對於治療產生的抗性與其幹細胞特性緊密相關。也因肝癌幹細胞具有自我更新及轉移的能力，可針對這些標的發展出有效的癌症治療。在本篇研究中，為了評估藥物的效力，使用鴨膽子的水溶液萃取物作用於三株肝癌細胞株，分別是HepG2, Huh7與Hep3B細胞。MTT試驗和流式細胞儀分析結果顯示萃取物會引起Hep3B細胞的sub-G1細胞總數表現且隨劑量增加有效抑制其生長。然而HepG2和Huh7細胞對於這處理較不敏感。西方墨點法證明了鴨膽子引起Hep3B細胞的凋亡之機制。再者，我們更進一步證實鴨膽子降低Hep3B spheroids自我更新能力，且能夠有效抑制幹細胞標記物。此外，水萃取物經由標的表皮生長因子受體而提升Hep3B spheroids細胞的凋亡，而自我更新能力的降低與幹細胞的凋亡有關。由這些結果證明鴨膽子水溶液萃取物可分別引發monolayer及spheroid的Hep3B細胞凋亡，這樣子的結果為肝癌的治療提供一種低劑量中藥的治療方式。

The Chinese herbal medicines (CHM), as one of the main natural resources, have been known in treating cancer. Their advantages include lower side effect, and angiogenesis and metastasis inhibition. Hepatocellular carcinoma (HCC) is one of the common malignant human tumors throughout Southeast Asia. HCC resisting to therapies are closely correlated with their stemness characteristics. Liver cancer stem-like cells have the ability of self-renewal and metastasis that have become the target for developing effective cancer therapy. In this study, an aqueous extract from Brucea javanica (BJ) was used to assess their effectiveness toward human HCC cells, HepG2, Huh7 and Hep3B cells. The results of MTT assay and Flow cytometry analysis showed that the extract inhibited cell growth in dose-dependent manner by inducing sub-G1 cell population in Hep3B. While HepG2 and Huh7 cells were insensitive to treatment. Western blot analysis demonstrated that the mechanisms of BJ induced apoptotic cell death in Hep3B cells. Furthermore, we also proved that BJ attenuated the self-renewal abilities of Hep3B spheroids and suppressed the stemness markers effectively. In addition, the extract augmented apoptotic characteristics of Hep3B spheroids by targeting epidermal growth factor receptor and the reduced self-renewal capacity was associated with apoptosis of stem cells. Taken together, the results suggested that aqueous BJ extract induced apoptosis in monolayer and the spheroids of Hep3B cells that provides a new alternative in treating a subset of liver cancer.

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