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研究生:林婷婷
研究生(外文):Ting-Ting Lin
論文名稱:探討牛樟芝CCM111對細胞訊息傳遞之影響
論文名稱(外文):The Study of CCM111 in Cell Signaling Transduction Pathways
指導教授:馬念涵
指導教授(外文):Nianhan Jia-Lin Ma
學位類別:碩士
校院名稱:國立中央大學
系所名稱:系統生物與生物資訊研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:英文
論文頁數:69
中文關鍵詞:發炎反應中草藥牛樟芝訊息傳遞
外文關鍵詞:inflammationsignaling transduction pathwayAntrodia camphorateTraditional chinese medicine
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牛樟芝,是台灣菌類特有種。一直以來被台灣原住民作為解酒以及治療肝病
的中草藥。有研究報導指出,牛樟芝萃取物具有抗肝炎病毒的效果並且能夠抑制
肝癌的形成。
訊息傳遞控制了細胞的功能,如細胞增生、分化等。而我們知道,許多癌症的產生與訊息傳遞的異常表現有關係。因此欲建立訊息傳遞平台,進行對牛樟芝的全面性篩選,進而得知牛樟芝能透過調控哪一條訊息傳遞而達到抗發炎反應甚至抗腫瘤生長的功能,為我實驗的主要目的。

CCM111, an extraction of Antrodia camphorate (AC) is a genus of fungi in Taiwan. It is well known as a Traditional Chinese Medicine (TCM) and has been widely used during Taiwanese aborigines as a drug for treating hangover and liver diseases.
In my study, we have collected stable cell lines expressing transcriptional regulatory elements in front of luciferase reporter to monitor activity of the signaling pathways.

摘要 . i
Abstract ii
誌謝 iii
Table of contents . iv
List of figures. vi
List of tables . viii
I. INTRODUCTION . 1
1. Antrodia camphorata . 1
2. Signaling transduction pathway 2
3. Inflammation-related signaling transduction pathway 2
4. Cytokine-related signaling transduction pathway . 4
5. Hormone related signaling transduction pathway 6
II. MATERIAL AND METHOD . 8
1. Materials . 8
1.1 Chemicals . 8
1.2 Antibodies . 8
1.3 Cell lines . 8
1.4 Stable clones with TRE luciferase reporters 9
1.5 pGL4.20 [luc2/Puro] . 9
1.6 Restriction enzymes 9
1.7 Primer sequences 9
1.8 DNA ligation enzyme . 10
2. Method 10
2.1 Establishment of stable clone expressing TRE luciferase reporter 10
v
2.2 Luciferase assay 10
2.3 Transformation . 11
2.4 Plasmid extraction 11
2.5 Transfection 11
2.6 SDS-PAGE and Western blot analysis . 11
2.7 Alamar blue assay . 12
III. RESULTS 13
1. Establishment of the luciferase assay platform for screening signaling
transduction pathways . 13
2. Establishment of Nrf2 luciferase reporter cell line . 13
3. Establishment of FOXO luciferase reporter cell line 14
4. Establishment of ELK-1 luciferase reporter cell line . 14
5. Titration of established luciferase reporter cell lines 15
6. The effect of CCM111 in signaling transduction pathways . 16
7. The toxicity of CCM111 in human mononuclear cells . 16
8. The effect of CCM111 in STAT3 signaling pathway . 17
9. Cell proliferation rate for CCM111 on colon cancer cell lines. 17
IV. DISCUSSION . 18
1. The effect of CCM111 in signaling transduction pathways . 18
2. CCM111 plays anti-inflammatory role . 19
3. Other functions of Antrodia camphorata 19
4. Future aspect . 20
V. REFERENCES . 21
FIGURES 23
TABLES . 55
1.Ao, Z.H., et al., Niuchangchih (Antrodia camphorata) and its potential in treating liver diseases. J Ethnopharmacol, 2009. 121(2): p. 194-212.
2.Phuong do, T., et al., Inhibitory effects of antrodins A-E from Antrodia cinnamomea and their metabolites on hepatitis C virus protease. Phytother Res, 2009. 23(4): p. 582-4.
3.Corvinus, F.M., et al., Persistent STAT3 activation in colon cancer is associated with enhanced cell proliferation and tumor growth. Neoplasia, 2005. 7(6): p. 545-55.
4.Levy, D.E. and C.-k. Lee, What does Stat3 do? J Clin Invest, 2002. 109(9): p. 1143-1148.
5.Scheidereit, C., IκB kinase complexes: gateways to NF-κB activation and transcription. Oncogene, 2006. 25(51): p. 6685-6705.
6.Klein, U. and S. Ghosh, The Two Faces of NF-ºB Signaling in Cancer Development and Therapy. Cancer Cell, 2011. 20(5): p. 556-558.
7.Kim, J., Y.-N. Cha, and Y.-J. Surh, A protective role of nuclear factor-erythroid 2-related factor-2 (Nrf2) in inflammatory disorders. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 2010. 690(1–2): p. 12-23.
8.Akira, S. and K. Takeda, Toll-like receptor signalling. Nat Rev Immunol, 2004. 4(7): p. 499-511.
9.Montagut, C. and J. Settleman, Targeting the RAF-MEK-ERK pathway in cancer therapy. Cancer Lett, 2009. 283(2): p. 125-34.
10.Zhang, W. and H.T. Liu, MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res, 2002. 12(1): p. 9-18.
11.Wickenden, J.A. and C.J. Watson, Key signalling nodes in mammary gland development and cancer. Signalling downstream of PI3 kinase in mammary epithelium: a play in 3 Akts. Breast Cancer Res, 2010. 12(2): p. 202.
12.Meulmeester, E. and P. ten Dijke, The dynamic roles of TGF-β in cancer. The Journal of Pathology, 2011. 223(2): p. 206-219.
13.Eivers, E., L.C. Fuentealba, and E.M. De Robertis, Integrating positional information at the level of Smad1/5/8. Current Opinion in Genetics & Development, 2008. 18(4): p. 304-310.
14.Nohe, A., et al., Signal transduction of bone morphogenetic protein receptors. Cellular Signalling, 2004. 16(3): p. 291-299.
15.Dahmani, R., P.-A. Just, and C. Perret, The Wnt/β-catenin pathway as a therapeutic target in human hepatocellular carcinoma. Clinics and Research in Hepatology and Gastroenterology, 2011. 35(11): p. 709-713.
16.Klaus, A. and W. Birchmeier, Wnt signalling and its impact on development and cancer. Nat Rev Cancer, 2008. 8(5): p. 387-98.
17.Oving, I.M. and H.C. Clevers, Molecular causes of colon cancer. European Journal of Clinical Investigation, 2002. 32(6): p. 448-457.
18.Chang, C.-y., et al., The Metabolic Regulator ERR±, a Downstream Target of HER2/IGF-1R, as a Therapeutic Target in Breast Cancer. Cancer Cell, 2011. 20(4): p. 500-510.
19.Stormshak, F. and C.V. Bishop, BOARD-INVITED REVIEW: Estrogen and progesterone signaling: Genomic and nongenomic actions in domestic ruminants. Journal of Animal Science, 2008. 86(2): p. 299-315.
20.Adams, J.S. and M. Hewison, Unexpected actions of vitamin D: new perspectives on the regulation of innate and adaptive immunity. Nat Clin Pract End Met, 2008. 4(2): p. 80-90.
21.White, J.H., Vitamin D Signaling, Infectious Diseases, and Regulation of Innate Immunity. Infection and Immunity, 2008. 76(9): p. 3837-3843.
22.Yang, H.-L., et al., Anti-metastatic activities of Antrodia camphorata against human breast cancer cells mediated through suppression of the MAPK signaling pathway. Food and Chemical Toxicology, 2011. 49(1): p. 290-298.
23.Rao, Y.K., et al., Identification of Antrocin from Antrodia camphorata as a Selective and Novel Class of Small Molecule Inhibitor of Akt/mTOR Signaling in Metastatic Breast Cancer MDA-MB-231 Cells. Chem Res Toxicol, 2010. 24(2): p. 238-245.
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