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研究生:何美儀
研究生(外文):Mei-I Ho
論文名稱:扁桃斑鳩菊萃取物誘導肝癌細胞凋亡及抑制癌細胞轉移之研究
論文名稱(外文):Vernonia amygdalina extracts induce apoptosis and inhibit EMT in human hepatocellular carcinoma cells
指導教授:高振益博士魏宗德博士
指導教授(外文):Jung-Yie KaoTzong-Der Way
口試委員:李正常博士
口試委員(外文):Jang-Chang Lee
口試日期:2016-07-20
學位類別:碩士
校院名稱:國立中興大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2016
畢業學年度:104
語文別:中文
論文頁數:51
中文關鍵詞:扁桃斑鳩菊肝癌
外文關鍵詞:Vernonia amygdalinahuman hepatocellular carcinoma cells
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肝癌這幾年來是台灣十大癌症死因的第二名,僅次於肺癌,每年奪走數千條寶貴生命。因肝臟是沉默的器官,當發現不適時,多已是晚期,而晚期肝癌的治療十分困難,因為大多數肝癌患者都合併有肝硬化或肝臟功能不佳,讓許多治療無法進行。所以找到肝癌的治療藥物是非常急迫的。在過去的研究中,藥用植物萃取物是治療癌症的明日之星,而扁桃斑鳩菊Vernonia amygdalina(VA)不僅具有抗氧化和抗發炎的能力,而且具有抗癌的特性,然而,關於VA的抗癌機制目前研究有限,本研究旨在探討肝臟上皮細胞癌(Hep 3B)在扁桃斑鳩菊萃取物作用下的抗癌效果及其作用機制。因萃取溶劑不同,萃取出來的成分亦會不同,首先用MTT assay初選出對肝癌細胞(Hep 3B)藥效較佳的萃取方式,結果從MTT分析得知,VA抑制Hep 3B的增殖,呈時間和劑量依賴性。接著流式細胞儀分析測試,在Hep 3B細胞中發現這種生長抑制的作用機制涉及細胞內特異性的G1 / S期的細胞週期停滯,進一步利用Propidine iodide和Annexin-V進行雙染實驗,證實VA會使Hep 3B細胞產生壞死。透過本研究證實VA確實有抗癌之功效,未來可望進一步發展成抗癌藥物。

Human hepatocellular carcinoma (HCC) is the second leading cancer-related cause of deaths in Taiwan. Due to fewer nerve distribution within liver, when the liver discomfort of symptoms, the disease often has already developed relatively serious, most patients with liver cancer or cirrhosis were associated with poor liver function, so that the treatment can''t be useful. So finding liver cancer treatment is very urgent. In past studies, medicinal plant extracts to treat cancer is growing more and more popular, and almond Vernonia Vernonia amygdalina (VA) has the ability to not only antioxidant and anti-inflammatory and anti-cancer properties, however, about the VA anticancer mechanism is finite, the present study was to investigate liver cell carcinoma (Hep 3B) anti-cancer effect and its mechanism in the almond extract Vernonia role. Vary the extraction solvent component extracted will also differ, first with the MTT assay primary efficacy preferred extraction methods on hepatoma cells (Hep 3B), the results from the analysis showed that MTT, VA to inhibit the proliferation of Hep 3B, in a time and dose-dependent manner. Followed by flow cytometric analysis and testing, we found that the mechanism involved in cell growth inhibition of cell cycle-specific G1 / S phase arrest in Hep 3B cells, further use Propidine iodide and Annexin-V double staining experiments confirmed VA will make Hep 3B cells necrosis. Through this study confirmed that VA does have anti-cancer effect, the future is expected to further develop anti-cancer drugs.

目錄

誌謝 i
中文摘要 iii
Abstract iv
目錄 v
圖表目次 vii
縮寫表 viii
第一章 前言 1
第一節、 癌症(Cancer) 1
第二節、 肝癌 3
一、 肝癌三部曲 3
二、 肝癌分類 3
三、 肝癌模式細胞 4
四、 肝癌致病因子 4
五、 肝癌臨床症狀 5
六、 肝癌臨床診斷 5
七、 肝癌臨床治療 6
第三節、 細胞週期(Cell Cycle) 7
第四節、 細胞程序性死亡 10
第五節、 細胞凋亡對癌細胞的分子機制 11
第六節、 上皮細胞間質轉換 14
第七節、 扁桃斑鳩菊 16
第八節、 研究動機與目的 18
第二章 材料與方法 19
第一節、 實驗材料 19
一、 細胞株 19
二、 試劑 19
三、 抗體 20
第二節、 主要儀器與耗材 20
第三節、 實驗用溶液配方 21
第四節、 實驗方法 25
一、 實驗藥物 25
二、 細胞培養 26
三、 細胞存活率分析MTT assay 27
四、 Colony formation的試驗 28
五、 細胞週期分析 28
六、 流式細胞儀之細胞死亡測試 29
七、 蛋白質萃取 30
八、 蛋白質的定量 30
九、 蛋白質之電泳分析(Sodium dodecyl sulfate polyacrylamide gel electrophoresis;SDS-PAGE) 31
十、 蛋白質轉印(Transfer)與影像呈現 31
十一、 細胞爬行能力試驗(Wound Healing Assaysv) 32
第三章 結果 33
第一節、 扁桃斑鳩菊對於肝癌細胞之細胞存活率測試 33
第二節、 扁桃斑鳩菊對於肝癌細胞之增生率測試 36
第三節、 扁桃斑鳩菊對於肝癌細胞的細胞週期測試 37
第四節、 扁桃斑鳩菊對於肝癌細胞的細胞死亡測試 39
第五節、 扁桃斑鳩菊誘導肝癌細胞凋亡路徑 蛋白質分析 41
第六節、 扁桃斑鳩菊抑制肝癌細胞爬行能力之分析 43
第七節、 扁桃斑鳩菊抑制肝癌細胞轉移相關因子表現之分析 45
第四章 討論與結論 47
第五章 參考文獻 49



1.Hanahan D and Weinberg RA. The hallmarks of cancer. cell. 2000; 100(1):57-70.
2.許金川. 疼惜阮的肝~肝炎、肝硬化、肝癌三部曲. 聲洋防癌之聲. 2007; (116):7-11.
3.Kudo M. Multistep human hepatocarcinogenesis: correlation of imaging with pathology. Journal of gastroenterology. 2009; 44(19):112-118.
4.Abdel-Hamid N. Update to risk factors for hepatocellular carcinoma. International Journal of Medicine and Medical Sciences. 2009; 1(3):038-043.
5.Molmenti EP, Marsh JW, Dvorchik I, Oliver JH, Madariaga J and Iwatsuki S. Hepatobiliary malignancies: primary hepatic malignant neoplasms. Surgical Clinics of North America. 1999; 79(1):43-57.
6.Kumar M, Zhao X and Wang XW. Molecular carcinogenesis of hepatocellular carcinoma and intrahepatic cholangiocarcinoma: one step closer to personalized medicine? Cell & bioscience. 2011; 1(1):1.
7.Feng H, Cheng AS, Tsang DP, Li MS, Go MY, Cheung YS, Zhao G-j, Ng SS, Lin MC and Yu J. Cell cycle–related kinase is a direct androgen receptor–regulated gene that drives β-catenin/T cell factor–dependent hepatocarcinogenesis. The Journal of clinical investigation. 2011; 121(8):3159-3175.
8.Çakir Ö, Topal U, Bayram AS and Tolunay S. Sarcomas: rare primary malignant tumors of the thorax. Diagnostic and Interventional Radiology. 2005; 11(1):23.
9.Dürr R and Caselmann WH. Carcinogenesis of primary liver malignancies. Langenbeck''s Archives of Surgery. 2000; 385(3):154-161.
10.Pineau P, Marchio A, Battiston C, Cordina E, Russo A, Terris B, Qin L-X, Turlin B, Tang Z-Y and Mazzaferro V. Chromosome instability in human hepatocellular carcinoma depends on p53 status and aflatoxin exposure. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2008; 653(1):6-13.
11.Poirier MC. Chemical-induced DNA damage and human cancer risk. Nature Reviews Cancer. 2004; 4(8):630-637.
12.Abraham RT. Cell cycle checkpoint signaling through the ATM and ATR kinases. Genes & development. 2001; 15(17):2177-2196.
13.Wiemels JL, Wiencke JK, Patoka J, Moghadassi M, Chew T, McMillan A, Miike R, Barger G and Wrensch M. Reduced immunoglobulin E and allergy among adults with glioma compared with controls. Cancer research. 2004; 64(22):8468-8473.
14.Sgambato A, Cittadini A, Faraglia B and Weinstein IB. Multiple functions of p27Kip1 and its alterations in tumor cells: a review. Journal of cellular physiology. 2000; 183(1):18-27.
15.Goldstein JC, Waterhouse NJ, Juin P, Evan GI and Green DR. The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Nature cell biology. 2000; 2(3):156-162.
16.MURAKAMI H and NURSE P. DNA replication and damage checkpoints and meiotic cell cycle controls in the fission and budding yeasts. Biochemical Journal. 2000; 349(1):1-12.
17.Glover DM, Leibowitz MH, McLean DA and Parry H. Mutations in aurora prevent centrosome separation leading to the formation of monopolar spindles. Cell. 1995; 81(1):95-105.
18.Malumbres M and Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nature Reviews Cancer. 2009; 9(3):153-166.
19.Sherr CJ. The Pezcoller lecture: cancer cell cycles revisited. Cancer research. 2000; 60(14):3689-3695.
20.Lloyd RV, Erickson LA, Jin L, Kulig E, Qian X, Cheville JC and Scheithauer BW. p27 kip1: a multifunctional cyclin-dependent kinase inhibitor with prognostic significance in human cancers. The American journal of pathology. 1999; 154(2):313-323.
21.Spirin KS, Simpson JF, Takeuchi S, Kawamata N, Miller CW and Koeffler HP. p27/Kip1 mutation found in breast cancer. Cancer Research. 1996; 56(10):2400-2404.
22.Hanahan D and Weinberg RA. Hallmarks of cancer: the next generation. cell. 2011; 144(5):646-674.
23.Majno G and Joris I. Apoptosis, oncosis, and necrosis. An overview of cell death. The American journal of pathology. 1995; 146(1):3.
24.Arends M, Morris R and Wyllie A. Apoptosis. The role of the endonuclease. The American journal of pathology. 1990; 136(3):593.
25.Zamzami N, Hirsch T, Dallaporta B, Petit PX and Kroemer G. Mitochondrial implication in accidental and programmed cell death: apoptosis and necrosis. Journal of bioenergetics and biomembranes. 1997; 29(2):185-193.
26.Eum K-H and Lee M. Crosstalk between autophagy and apoptosis in the regulation of paclitaxel-induced cell death in v-Ha-ras-transformed fibroblasts. Molecular and cellular biochemistry. 2011; 348(1-2):61-68.
27.Shen H-M and Pervaiz S. TNF receptor superfamily-induced cell death: redox-dependent execution. The FASEB Journal. 2006; 20(10):1589-1598.
28.Kang J, Kisenge RR, Toyoda H, Tanaka S, Bu J, Azuma E and Komada Y. Chemical sensitization and regulation of TRAIL‐induced apoptosis in a panel of B‐lymphocytic leukaemia cell lines. British journal of haematology. 2003; 123(5):921-932.
29.Boldin MP, Varfolomeev EE, Pancer Z, Mett IL, Camonis JH and Wallach D. A novel protein that interacts with the death domain of Fas/APO1 contains a sequence motif related to the death domain. Journal of Biological Chemistry. 1995; 270(14):7795-7798.
30.Zimmermann KC, Bonzon C and Green DR. The machinery of programmed cell death. Pharmacology & therapeutics. 2001; 92(1):57-70.
31.Adams JM and Cory S. The Bcl-2 protein family: arbiters of cell survival. Science. 1998; 281(5381):1322-1326.
32.Garrido C, Galluzzi L, Brunet M, Puig P, Didelot C and Kroemer G. Mechanisms of cytochrome c release from mitochondria. Cell Death & Differentiation. 2006; 13(9):1423-1433.
33.Cadenas E and Davies KJ. Mitochondrial free radical generation, oxidative stress, and aging. Free Radical Biology and Medicine. 2000; 29(3):222-230.
34.Bell EL, Klimova TA, Eisenbart J, Moraes CT, Murphy MP, Budinger GS and Chandel NS. The Qo site of the mitochondrial complex III is required for the transduction of hypoxic signaling via reactive oxygen species production. The Journal of cell biology. 2007; 177(6):1029-1036.
35.Tee T-T, Cheah Y-H, Meenakshii N, Sharom MYM and Hawariah LPA. Xanthorrhizol induced DNA fragmentation in HepG2 cells involving Bcl-2 family proteins. Biochemical and biophysical research communications. 2012; 420(4):834-838.
36.MacFarlane M and Williams AC. Apoptosis and disease: a life or death decision. EMBO reports. 2004; 5(7):674-678.
37.Thiery JP. Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002; 2(6):442-454.
38.Lee JM, Dedhar S, Kalluri R and Thompson EW. The epithelial-mesenchymal transition: new insights in signaling, development, and disease. The Journal of cell biology. 2006; 172(7):973-981.
39.Yedjou CG, Izevbigie EB and Tchounwou PB. Vernonia amygdalina—Induced Growth Arrest and Apoptosis of Breast Cancer (MCF-7) Cells. Pharmacology & pharmacy. 2013; 4(1).
40.Kupchan SM, Hemingway RJ, Karim A and Werner D. Tumor inhibitors. XLVII. Vernodalin and vernomygdin, two new cytotoxic sesquiterpene lactones from Vernonia amygdalina Del. The Journal of organic chemistry. 1969; 34(12):3908-3911.
41.Akinpelu DA. Antimicrobial activity of Vernonia amygdalina leaves. Fitoterapia. 1999; 70(4):432-434.
42.Akah P and Okafor C. Blood sugar lowering effect of Vernonia amygdalina Del, in an experimental rabbit model. Phytotherapy Research. 1992; 6(3):171-173.



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