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研究生:陳怡甄
研究生(外文):Yi-Chen Chen
論文名稱:探討從萃取自石蓮花之高活性分子對抗肝癌的效用
論文名稱(外文):Characterization of Active molecules extracted from Graptopetalum paraguayense with Anti-Cancer Activity in Hepatocellular Carcinoma
指導教授:黃奇英
指導教授(外文):Chi-Ying F. Huang
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物藥學研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:英文
論文頁數:60
中文關鍵詞:中草藥肝細胞癌
外文關鍵詞:Chinese herbal medicineHepatocellular carcinomaZC008
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肝細胞癌 (Hepatocellular carcinoma,簡稱 HCC) 在原發性肝癌中是最常見的一種類型,不只在常見的惡性腫瘤中排名第六位,更是在全球性癌症死亡上排名第三位。在臨床治療上,肝細胞癌因高度惡性化及症狀常在晚期發生不易早期偵察,預後一般很差。目前已知肝纖維化和肝硬化皆為造成肝細胞癌的主要危險因子之一。為了找尋有效治療肝細胞癌藥物,我們從中草藥著手,發現一中草藥 (石蓮花)能有效抑制大白鼠經由二甲基亞硝胺 (DMN) 所誘發之肝纖維化及發炎反應,並且會調控活化態的肝星狀細胞 (HSC-T6細胞株) 中致癌基因的蛋白質表現量。石蓮花亦抑制了 Huh7 和 HepG2 細胞株的細胞生長及致癌基因的蛋白質表現量 (AURKA,AURKB,和FLJ10540)。石蓮花 經過進一步的純化及透過活性測試而得到含有活性成份的第三個組分(HH-F3)。同時,石蓮花和HH-F3與具有抑制 AURKA 活性的 aurora kinase 抑制劑 (VE465) 相比,在抑制AURKA 的蛋白質表現量上具有更好的效用。而HH-F3在48小時作用於 Huh7,PLC5,及 Mahlavu 三種細胞株的 IC50 分別為50, 100, 與20 μg/ml。此外,HH-F3可能透過調控磷酸化的 JNK,ERK 和 Akt 訊息傳遞途徑而造成 HCC 的細胞株走向細胞凋亡。綜合上述,HH-F3在治療肝細胞癌的效果上提供一個新的解決方案。
Hepatocellular carcinoma (HCC) is the sixth most common malignancy and the third most common cause of cancer deaths globally. Both hepatic fibrosis and cirrhosis are the risk factors for hepatocellular carcinoma. To identify the key therapeutic agents, we previously found that one of the Chinese herbal medicines, Graptopetalum paraguayense, can prevent DMN-induced hepatic inflammation and fibrosis in animal model and down-regulate the protein expression of several oncogenes in HSC-T6 cells. This observation raises the possibility that Graptopetalum paraguayense might have anti-cancer activity and we might use this down-regulation as a criterion for the purification of active components from Graptopetalum paraguayense. Graptopetalum paraguayense results in cytotoxicity effects and down-regulates AURKA, AURKB, and FLJ10540 expression in Huh7 and HepG2 cells. HH-F3, the fraction with active components, suppressed the protein expression of those above mentioned proteins. Unlike VE-465 (Aurora kinase inhibitor), Graptopetalum paraguayense and HH-F3 could not inhibit histone H3 phosphorylation on Ser10. The IC50 for HH-F3 at 48 hours on Huh7, PLC5, and Mahlavu cells was 50, 100, and 20 μg/ml, respectively. In addition, HH-F3 induced HCC to undergo apoptosis through down-regulation of JNK, ERK, and Akt pathway. In conclusion, HH-F3 was demonstrated the potential therapeutic effects on the treatment of HCC and liver fibrosis.
Content

Abstract 3
Chinese Abstract 4
Introduction 5
Liver Fibrosis 5
Hepatocellular Carcinoma (HCC) 6
Graptopetalum paraguayense and HH-F3 8
Aurora Kinase 10
FLJ10540 (CEP55) 10
Cell proliferation 11
Cell Death in Tumor Cells 12
Specific Aims 13
Materials and Methods 14
Results 20
The effects of Graptopetalum paraguayense on interphase and mitotic Huh7/HepG2 cells 20
Different purification fractions of Graptopetalum paraguayense have distinct effects on protein expression in HCC cell lines 20
HH-F3 down-regulates AURKA and FLJ10540 and has different effect from VE-465 in liver cancer cells 21
AURKA protein is down-regulated under HH-F3 treatment, whereas there is no change at the transcription level 22
HH-F3can inhibit cell viability on HCC cell lines 23
HH-F3 induces apoptosis or necrosis in HCC cell lines 23
HH-F3 leads to cell death through apoptosis in HCC cell lines 24
HH-F3 decreases mitochondrial membrane potential in HCC cell lines 25
The protein expression of SOD1 was increased under HH-F3 treatment in HCC cell lines 25
HH-F3 suppresses the phosphorylation of Akt, JNK1/2, and ERK1/2 in HCC cells 26
Discussion 27
Reference 32
Figures 38
Appendix 53


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