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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林榮耀(Jung-Yaw Lin) | |
| dc.contributor.author | Kang-Hsu Fan | en |
| dc.contributor.author | 范綱緒 | zh_TW |
| dc.date.accessioned | 2021-06-16T10:20:07Z | - |
| dc.date.available | 2018-09-24 | |
| dc.date.copyright | 2013-09-24 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-16 | |
| dc.identifier.citation | 1 El-Serag, H. B. Hepatocellular carcinoma. The New England journal of medicine 365, 1118-1127, doi:10.1056/NEJMra1001683 (2011).
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Y., Yoon, S. O., Kim, S. G., Roux, P. P. & Blenis, J. Rapamycin differentially inhibits S6Ks and 4E-BP1 to mediate cell-type-specific repression of mRNA translation. Proceedings of the National Academy of Sciences of the United States of America 105, 17414-17419, doi:10.1073/pnas.0809136105 (2008). 56 Pang, X. et al. 1'-Acetoxychavicol acetate suppresses angiogenesis-mediated human prostate tumor growth by targeting VEGF-mediated Src-FAK-Rho GTPase-signaling pathway. Carcinogenesis 32, 904-912, doi:10.1093/carcin/bgr052 (2011). 57 Yang, H. et al. mTOR kinase structure, mechanism and regulation. Nature 497, 217-223, doi:10.1038/nature12122 (2013). 58 Stefania Di, T. et al. Structure of genipin in solution: a combined experimental and theoretical study. RSC Advances 3, doi:10.1039/c3ra42147c (2013). 59 Ran, T. et al. A selectivity study on mTOR/PI3Kalpha inhibitors by homology modeling and 3D-QSAR. Journal of molecular modeling 18, 171-186, doi:10.1007/s00894-011-1034-3 (2012). | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60508 | - |
| dc.description.abstract | 肝細胞癌(Hepatocellular carcinoma)是全球發生率及死亡率極高的癌症之一，在台灣也高居十大癌症死亡原因的第二名。可能造成肝細胞癌形成的致病機轉有很多，如長期感染B型或C型肝炎病毒、酒精性肝炎以及長期攝取受黃麴毒素感染的食物等等。目前治療肝細胞癌最有效的方式是手術切除，然後針對那些無法經由手術治療的患者，化學療法成為另一個選擇。目前在臨床上已經有許多小分子藥物被開發，並且應用於肝細胞癌的治療，像是Sorafenib。但由於肝癌細胞具有高度轉移性，導致復發率居高不下。基於此點，研發抗肝細胞癌轉移的藥物成為一個急切的課題。
科學界近年積極探究中草藥的治病機轉，探索中藥治療肝癌之路。本研究以Huh7肝細胞癌細胞株來檢測中草藥對於抗肝細胞癌之效果。在篩選多數中草藥之後，發現NTNU05可以不毒殺細胞的方式抑制細胞爬行。進一步分析此抗細胞轉移的細胞機制，發現NTNU05可以透過阻礙FAK/Src之訊息傳遞路徑而抑制Rac1的活化，進而抑制肌絲蛋白重組，最後導致Huh7細胞爬行能力下降。NTNU05亦可抑制AKT/mTOR之訊息傳遞路徑，最終導致VEGFA的表現量減少而抑制血管新生。此外，NTNU05可以穩定E-cadherin和beta-catenin之複合體，並抑制Snail以及Slug的進核，而抑制Huh7細胞轉變為間葉細胞(mesenchyme)型態。本研究也利用異種移植(xenograft)腫瘤小鼠模式，將Huh7細胞注射到嚴重免疫不全症小鼠(NOD-SCID)之皮下，探討NTNU05在活體(in vivo)的抗癌效果。發現NTNU05可以顯著抑制腫瘤生長;除此之外，透過免疫組織化學染色法鑑定(IHC)，CD31(血管上皮之標記)亦顯著減少，顯示NTNU05在活體中亦可透過抑制血管新生之方式進而抑制腫瘤生長。 本研究亦探究NTNU05有效成分之-NTNU05-1之抗癌能力。發現NTNU05-1亦可以透過抑制Rho GTPases和mTOR相關訊息傳遞路徑而達到抑制Huh7細胞之爬行和血管新生。根據以上的實驗結果，本研究證明中草藥NTNU05以及其有效成分-NTNU05-1具有抑制肝細胞癌轉移及血管新生的能力，在往後肝細胞癌的治療甚至預防上，具有相當大的發展潛力。 | zh_TW |
| dc.description.abstract | Hepatocellular carcinoma (HCC) is one of the most malignant human cancers over the world, and now, it is the second cause of cancer deaths in Taiwan. It is usually caused by hepatitis virus infection, alatoxin-B1-contained food intake, and chromic alcohol consumption. Multiple therapeutic strategies have been developed so far, such as some small molecular compounds like Sorafinib, which is used in clinical treatment. However, the cure rate of HCC remains poor due to the high metastatic effect with the high recurrence rate. Therefore. it is an urgent need for investigation of a novel anti-metastasis drug for therapy of HCC.
Recently, Chinese Herbal Medicines (CHMs) are wildly used on the prevention and treatment against HCC. In this study, we found that CHM NTNU05 water-extracts, which are usually used to treat rheumatoid arthritis by its anti-inflammatiotory activity has an inhibitory effect on Huh7 cells migration, demonstrated by transwell migration/invasion assays. We further showed that aqueous extracts of NTNU05 could inhibit Huh7 cells migration/invasion by suppressing the FAK/SRC/Rac1, resulting in disrupting the F-actin rearrangement. NTNU05 also inhibited AKT/mTOR pathway, leading to the decrement of VEGFA. In addition, aqueous extracts of NTNU05 could promote the formation of E-cadherin and beta-catenin complex, the adhesive structures between adjacent cells, and decreased the translocation of Snail/Slug to nucleus. We further showed that NTNU05 also had inhibitory effects on tumor growth and angiogenesis in vivo by a mouse xenograft model. Furthermore, we found that NTNU05-1, one of the active components of NTNU05, contributed its anti-migration and angiogenesis activities to NTNU05 through inhibition of RhoGTPases and mTOR-related pathway. Taken together, NTNU05 and NTNU05-1 may be potential chemotherapeutic agents for HCC. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T10:20:07Z (GMT). No. of bitstreams: 1 ntu-102-R00442005-1.pdf: 7209329 bytes, checksum: ad8f3ac98ee67b984a9b2d05313e2f58 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 摘要 I
Abstract III Chapter 1. Introduction 1 1. Hepatocellular cell carcinoma (HCC) 2 2. NTNU05 3 3. NTNU05-1 3 4. VEGF pathway 4 5. Mammalian target of rapamycin (mTOR) 5 6. Rho-GTPase family 7 7. Research purpose of present investigation 8 Chapter 2. Materials and methods 9 1. Cell lines, Animals, and Reagents 10 2. Preparation of Chinese Herb Medicine (CHM) 11 3. Cell culture 11 4. Cell viability assay 11 5. Cell migration/ invasion assay 12 6. Wound healing assay 13 7. Confocal microscopy 13 8. RNA extraction and reverse transcription 14 i. RNAs extraction 14 ii. Reverse transcription 15 9. Real time-quantitative PCR (Q-PCR) 16 10. Western blot analysis 16 i. Preparation of cell lysates: 16 ii. Quantification of protein concentration 17 iii. Preparation of sodium-dodecyl-sulfate –polyacrylamide gels (SDS-polyacrylamide gels) 18 iv. Protein sample preparation 19 v. Electrophoresis 19 vi. Semi-dry blotting 20 vii. Immunoblotting 20 11. Nuclear and cytoplasmic fractionation 21 12. GST-PBD/RBD pull down assay 22 i. Purification of GST-PBD (p21-binding domain, rac1/cdc42 binding domain) and GST-RBD (Rho binding domain) from E.coli extracts 22 ii. Cdc42/Rac1 and RhoA activity assay 23 13. In vitro angiogenesis: endothelial cell tube formation assay 24 14. Tumor xenograft analysis for tumor growth 24 15. 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay for measurement of ROS scavenging activity 25 16. Chemiluminescence assay for ROS measurement 26 17. Statistical analysis 26 Chapter 3. Results 27 1. NTNU05 showed inhibitory effects on migration/invasion of Huh7 cells in a non-toxic manner. 28 2. NTNU05 disrupted the actin rearrangement of Huh7 cells by inhibiting Rac-1 activity via Src/FAK signaling. 28 3. NTNU05 inhibited epithelial-mesenchymal transition (EMT) of Huh7 cells. 29 4. NTNU05 suppressed AKT/mTOR/HIF1a/VEGFA signaling pathway. 30 5. NTNU05 inhibited VEGF-induced migration and capillary structure formation of Human umbilical vein endothelial cells (HUVECs). 32 6. NTNU05 down-regulated ROS production of Huh7 cells. 32 7. NTNU05 decreased tumor growth and angiogenesis in vivo. 33 8. NTNU05-1 inhibited Huh7 cell migration in non-toxic manner. 34 9. NTNU05-1 suppressed mTOR signaling pathway. 35 10. NTNU05-1 inhibited VEGFR2 activation. 36 11. NTNU05-1 inhibited motility of Huh7 cells by suppressing activities of Rac1, Cdc42, and RhoA. 36 Chapter 4. Discussion 38 Chapter 5. Figures 44 Chapter 6. Table 75 Chapter 7. References 77 | |
| dc.language.iso | en | |
| dc.subject | 細胞爬行 | zh_TW |
| dc.subject | 血管新生 | zh_TW |
| dc.subject | 血管內皮生長因子-A | zh_TW |
| dc.subject | 中草藥 | zh_TW |
| dc.subject | 肝細胞癌 | zh_TW |
| dc.subject | mTOR | zh_TW |
| dc.subject | mTOR | en |
| dc.subject | Chinese Herbal Medicine | en |
| dc.subject | Migration | en |
| dc.subject | Angiogenesis | en |
| dc.subject | VEGFA | en |
| dc.subject | Hepatocellular carcinoma | en |
| dc.title | 中草藥: NTNU05抗肝癌之研究 | zh_TW |
| dc.title | Mechanisms of anti-hepatocellular carcinoma of Chinese Herbal Medicine: NTNU05 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李明學(Ming-Shyue Lee),呂紹俊(Shao-Chun Lu),李德章(Te-Chang Lee) | |
| dc.subject.keyword | 肝細胞癌,中草藥,細胞爬行,血管新生,血管內皮生長因子-A,mTOR, | zh_TW |
| dc.subject.keyword | Hepatocellular carcinoma,Chinese Herbal Medicine,Migration,Angiogenesis,VEGFA,mTOR, | en |
| dc.relation.page | 82 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2013-08-16 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 生物化學暨分子生物學研究所 | zh_TW |
| 顯示於系所單位： | 生物化學暨分子生物學科研究所 | |
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