臺灣博碩士論文加值系統

牛樟芝（學名Antrodia cinnamomea）是台灣特有的菌菇，具有抗發炎、抗氧化及抗血管增生等功能。台灣民間常用牛樟芝來治療肝癌、肺癌及乳癌等，但相關的研究及作用機制仍有許多不清楚之處。本研究在探討牛樟芝的純化合物Antrocin及Antcin H對於泌尿道的兩種癌症：膀胱癌及腎癌的作用及其機轉。結果發現高濃度Antrocin處理時，膀胱癌細胞會發生死亡現象，其機轉是透過提高Fas、DR5及Bax的表現和促進Caspases 3、 Caspases 8、及Caspases 9的活性，經由內在及外在路徑，造成細胞凋亡（apoptosis）。若將膀胱癌細胞處理以低濃度非致死劑量的Antrocin，可顯著地抑制癌細胞生長，移動（migration）和侵襲（invasion），其機轉為抑制磷酸化FAK（focal adhesion kinase）和磷酸化paxillin的活化。同時，可觀察到Antrocin可減少FAK 和paxillin在癌細胞邊緣的分佈，繼而抑制癌細胞產生偽足運動，從而降低其侵襲能力。另外發現Antrocin會提昇癌細胞上皮附著分子E-cadherin的表現和減少Vimentin的表現，進一步更觀察到Antrocin可降低膀胱癌細胞中基質金屬蛋白酶（matrix metalloproteinases, MMPs）的表現，其中又以MMP-2的表現受抑制最為明顯。另外發現以Antrocin處理的膀胱癌細胞中，磷酸化ERK和c-Fos表現量降低。染色質免疫沉澱法（chromatin immunoprecipitation assay）分析之結果，顯示Antrocin降低c-Fos與MMP-2上游DNA啟動位點上的結合能力，此結果可能就是Antrocin抑制MMP-2表現的機制之一。本研究論文中另一部分，是探討Antcin H處理腎細胞癌的機轉，發現Antcin H可抑制腎癌細胞生長，其處理48小時的IC50為170 μM。在非致死劑量（低於100 μM）的Antcin H處理下，細胞遷移和侵襲的能力也顯著降低，這種抑制作用，導因於抑制FAK和Src激酶活性，抑制paxillin磷酸化和vimentin的表現，進而破壞偽足形成，調昇TIMP家族的表現，並抑制MMP家族的表現，而其中又以MMP-7的表現量被抑制得最顯著。從螢光素酶報告基因檢測 (reporter luciferase assay)的數據表示，Antcin H除了抑制MMP-7的啟動子活性外，也同時抑制c-Fos/AP-1和C/EBP-β對MMP-7基因的轉錄活性。此外，Antcin H強烈抑制ERK1/2的活性，並且抑制癌細胞內的c-Fos/AP-1和C/EBP-β與MMP-7上游啟動子結合位點的結合力。總而言之，本論文之研究，是第一個發現Antrocin可抑制膀胱癌細胞的遷移和侵襲，以及Antcin H對人類腎癌細胞之抗遷移和抗侵襲作用。我們的發現提供Antrocin作為膀胱癌的替代療法，以及Antcin H可能有治療轉移性腎細胞癌的潛力之實證數據。

Antrodia cinnamomea, a famous medicinal mushroom, known as Niuchang chih in Taiwan, exhibits anti-inflammatory, anti-oxidative, anti-angiogenic and hepatic protection properties. Several in vitro studies demonstrate the anti-cancer effects of Antrodia cinnamomea on hepatoma, lung cancer, as well as breast cancer. In this study, the effects of two active compound, Antrocin and Antcin H isolated from Antrodia cinnamomea against urological malignancies, both bladder cancer and renal cell carcinoma (RCC) were examined. Our study showed that treatment with cytotoxic concentration of Antrocin induced both intrinsic and extrinsic apoptotic pathways in human bladder cancer 5637 cells, evidenced by increase of Fas, DR5, Bax expression and caspase-3, -8 and -9 activation. Exposure to non-cytotoxic concentrations of antrocin significantly inhibited cell growth, migration, and invasion, which was associated with decreased phosphorylation of focal adhesion kinase (FAK) and paxillin. Antrocin also reduced subcellular distribution of FAK and paxillin at the focal adhesion contacts of the cell periphery site, and disrupted the formation of filopodia and lamellipodia. Moreover, Antrocin increased epithelial-to-mesenchymal transition-related gene E-cadherin and decreased vimentin expression. Real time-PCR analysis showed that Antrocin downregulated the expression of mRNA of several matrix metalloproteinases (MMPs), including MMP-2. Also, the phosphorylation of ERK and c-Fos were attenuated by Antrocin. Data from chromatin immunoprecipitation assay demonstrated that antrocin decreased the DNA binding activity of c-Fos to the upstream/enhancer region of MMP-2 promoter, an action likely to result in reducing MMP-2 expression. Next, the anti-cancer effects of antcin H were investinged in human renal carcinoma 786-0 cells. The results showed that antcin H significantly inhibited the growth of RCC 786-0 cells, the IC50 value (for 48 h) of antcin H was 170 μM. Besides, the migration and invasion of 786-0 cells were also drastically suppressed by antcin H under non-cytotoxic concentrations (< 100 μM), these events were accompanied by the inhibition of FAK and Src kinase activities, decreases of paxillin phosphorylation and vimentin expressions, impairment of focal contact and lamellipodium formation, and up-regulation of tissue inhibitor of metalloproteinases (TIMPs) as well as down-regulation of several MMPs, especially MMP-7 expression. Data from the reporter luciferase assay showed that antcin H repressed the MMP-7 promoter activity, in parallel to inhibition of c-Fos/AP-1 and C/EBP-β transactivation properties. Moreover, antcin H strongly suppressed the activity of ERK1/2 and decreased the binding ability of C/EBP-β and c-Fos on the upstream/enhancer region of MMP-7 promoter. Overall, this is the first study demonstrates that Antrocin inhibited the migration and invasion of bladder cancer cells as well as the anti-migratory and anti-invasive effects of the Antcin H on RCC 786-0 cells. Our findings provide the evidence that Antrocin may be the substitutional therapy of bladder cancer, while Antcin H may have the potential for application on treating metastatic RCC.

論文摘要.............................................................................I
Abstract .........................................................................III
略字表...............................................................................V
誌謝................................................................................VI
目錄................................................................................VII
第一章 前言...........................................................................1
第二章 緒論...........................................................................3
第一節 膀胱泌尿上皮癌的簡介.............................................................3
第二節 腎細胞癌的簡介..................................................................4
第三節 細胞凋亡.......................................................................5
第四節 癌症的侵襲(Invasion)與轉移(Metastasis)..........................................6
第五節 牛樟芝的簡介....................................................................7
第三章 研究動機........................................................................9
第四章 材料與方法.....................................................................10
第一節 實驗材料......................................................................10
一、細胞株來源及培養條件..............................................................10
二、實驗藥劑及試劑...................................................................10
第二節 實驗方法......................................................................11
一、生長抑制測定.....................................................................11
二、胱天蛋白酶活性測定................................................................11
三、傷口癒合實驗.....................................................................12
四、遷移和侵襲實驗...................................................................12
五、酶譜分析........................................................................12
六、免疫印跡分析.....................................................................13
七、RNA的萃取和real time-PCR........................................................13
八、免疫螢光染色.....................................................................13
九、染色質免疫沉澱法.................................................................14
十、MMP-2啟動子結構和啟動子功能的測定..................................................15
十一、西方點墨法.....................................................................16
十二、螢光素酶檢測...................................................................16
第三節 統計分析法.....................................................................17
第五章 結果...........................................................................18
第一節 Antrocin 處理膀胱泌尿上皮癌之研究................................................18
第二節Antcin H 處理腎細胞癌之研究......................................................22
第六章 討論...........................................................................28
第七章 結論...........................................................................35
參考文獻.............................................................................37
圖表目錄
圖一、Antrocin的化學結構式............................................................49
圖二、Antrocin可抑制人類膀胱癌5637細胞之生長............................................49
圖三、Antrocin誘導細胞凋亡............................................................50
圖四、胱天蛋白酶可被Antrocin活化.......................................................50
圖五、Antrocin調控細胞凋亡之相關分子之表現...............................................51
圖六、Antrocin抑制傷口癒合............................................................51
圖七、Antrocin可抑制癌細胞的遷移能力....................................................52
圖八、Antrocin可阻礙癌細胞的侵襲力.....................................................52
圖九、Antrocin調控與轉移相關之分子的活性及表現...........................................53
圖十、Antrocin調控磷酸化FAK在癌細胞內之分布及表現........................................53
圖十一、Antrocin調控磷酸化PXN在癌細胞內之分布及表現...............54
圖十二、在Antrocin處理後膀胱癌細胞癒合區邊緣細胞之lamellipodia和絲狀偽足與磷酸化FAK的分布.....54
圖十三、在Antrocin處理後膀胱癌細胞癒合區邊緣細胞之lamellipodia和絲狀偽足與磷酸化PXN的分布.....55
圖十四、Antrocin調控MMP家族分子之mRNA表現..............................................55
圖十五、Antrocin降低MMP-2分解酵素之活性................................................56
圖十六、Antrocin抑制磷酸化FAK，磷酸化ERK及c-Fos之活性及表現...............................56
圖十七、位於MMP-2啟動區上游的c-Fos結合區的示意圖.........................................57
圖十八、Antrocin減少c-Fos與MMP-2啟動子上對應點之接合力...................................57
圖十九、Antrocin抑制MMP-2啟動子的活化..................................................58
圖二十、Antcin H的化學結構式..........................................................58
圖二十一、Antcin H可抑制腎癌細胞之生長..................................................59
圖二十二、Antcin H抑制腎癌細胞之轉移...................................................59
圖二十三、Antcin H抑制腎癌細胞轉移相關之分子的活性表現....................................60
圖二十四、Antcin H降低磷酸化FAK及lamellipodia的形成................................... 60
圖二十五、Antcin H可改變磷酸化PXN在腎細胞內分布之位置及表現量..............................61
圖二十六、Antcin H可抑制腎癌細胞之轉移及傷口癒合.........................................61
圖二十七、傷口癒合的磷酸化FAK活性及表現量可被Antcin H抑制.................................62
圖二十八、傷口前緣之磷酸化PXN之分布及表現受Antcin H抑制...................................62
圖二十九、Antcin H抑制腎癌細胞之侵襲作用................................................63
圖三十、Antcin H抑制腎癌細胞之侵襲作用..................................................64
圖三十一、Antcin H調控Scr，FAK及ERK之活性..............................................65
圖三十二、Antcin H抑制Scr，FAK及ERK訊號路徑之活性.......................................65
圖三十三、Antcin H抑制 MMP-7啟動子之活化...............................................66
圖三十四、坐落在MMP-7的起始端上游的c-Fos/ AP1和C/EBP-β的結合位點示意圖.....................66
圖三十五、c-Fos及C/EBP-β與MMP-7啟動子結合區之連接受Antcin H之阻礙.........................67
圖三十六、Antcin H以劑量依賴的型態，調降c-Fos與C/EBP-β之結合力............................68
圖三十七、Antrocin抑制膀胱癌細胞增殖和轉移作用的分子機制示意圖..............................69
圖三十八、Antrocin抑制膀胱癌細胞增殖和轉移作用的卡通繪圖...................................69
圖三十九、Antcin H抑制腎癌細胞增殖和轉移作用的分子機制示意圖................................70

1. Chen, B.K., and Yang, C.Y. “Temporal Trend Analysis of Avoidable Mortality in Taiwan 1971-2008: Overall Progress, with Areas for Further Medical or Public Health Investment,” BMC Public Health, Vol. 13, pp.551-582 (2013)
2. Yamaguchi, H., Wyckoff, J., and Condeelis, J., “ Cell Migration in Tumors,” Current Opinion in Cell Biology, Vol. 17, No. 5, pp.559-564 (2005)
3. Friedl, P. and Wolf, K., “Tumour-Cell Invasion and Migration: Diversity and Escape Mechanisms,” Nature Reviews Cancer, Vol. 3, No. 5, pp.362-374 (2003)
4. Hsia, D.A., Mitra, S.K., Hauck, C.R., Streblow, D.N., Nelson, J.A., Ilic, D., Huang, S., Li, E., Nemerow, G.R., Leng, J., Spencer, K.S.R., Cheresh, D.A., and Schlaepfer, D.D., “Differential Regulation of Cell Motility and Invasion by FAK,” The Journal of Cell Biology, Vol. 160, No. 5, pp.753-767 (2003)
5. Bagi, C.M., Roberts, G.W., and Andresen, C.J., “Dual Focal Adhesion Kinase/Pyk2 Inhibitor Has Positive Effects on Bone Tumors,” Cancer, Vol. 112, No. 10, pp.2313-2321 (2008)
6. Parkin, D.M., “The Global Burden of Urinary Bladder Cancer,” Scandinavian Journal of Urology and Nephrology. Supplementum, Vol. 218, pp.12-20 (2008)
7. Chu, H., Wang, M., and Zhang, Z., “Bladder Cancer Epidemiology and Genetic Susceptibility,” Journal of Biomedical Research, Vol. 27, No. 3, pp.170-178 (2013)
8. Montironi, R., and Lopez-Beltran, A., “The 2004 WHO Classification of Bladder Tumors: a Summary and Commentary,” International Journal of Surgical Pathology, Vol. 13, No. 2, pp.143-153 (2005)
9. Pillai, R., Wang, D., Mayer, E.K., and Abel, P., “Do Standardised Prognostic Algorithms Reflect Local Practice? Application of EORTC Risk Tables for Non-Muscle Invasive (pTa/pT1) Bladder Cancer Recurrence and Progression in a Local Cohort,” The Scientific World Journal, Vol. 5, No. 11, pp.751-759 (2011)
10. Zincke, H., Utz, DC., Taylor, W.F., Myers, R.P., and Leary, F.J., “Influence of Thiotepa and Doxorubicin Instillation at Time of Transurethral Surgical Treatment of Bladder Cancer on Tumor Recurrence: A Prospective, Randomized Double Blind, Controlled Trial,” The Journal of Urology, Vol. 129, pp.505-509 (1983)
11. Oddens, J.R., van der Meijden, A.P., and Sylvester, R., “One Immediate Postoperative Instillation of Chemotherapy in Low Risk Ta, T1 Bladder Cancer Patients. Is It Always Safe?,” European Urology, Vol. 46, pp.336-338 (2004)
12. Sternberg, C.N., and Collette, L., “What Has Been Learned from Meta-Analyses of Neoadjuvant and Adjuvant Chemotherapy in Bladder Cancer?,” BJU International, Vol. 98, No. 3, pp.487-489 (2006)
13. Chen, K.S., Lai, M.K., Huang, C.C., Chu, S.H., and Leu, M.L., “Urologic Cancers in Uremic Patients,” American Journal of Kidney Disease, Vol. 25, No. 5, pp.694-700 (1995)
14. Landis, S.H., Murray, T., Bolden, S., and Wingo, P.A., “Cancer Statistics: 1999,” CA: a Cancer Journal of Clinicians, Vol. 49, pp.8-31 (1999)
15. Pantuck, A.J., Seligson, D.B., Klatte, T., Yu, H., Leppert, J.T., Moore, L., O'Toole, T., Gibbons, J., Belldegrun, A.S., and Figlin, R.A., “Prognostic Relevance of the mTOR Pathway in Renal Cell Carcinoma,” Cancer, Vol. 109, No. 11, pp.2257-2267 (2007)
16. Lisztwan, J., Imbert, G., Wirbelauer, C., Gstaiger, M., and Krek, W., “The von Hippel-Lindau Tumor Suppressor Protein Is a Component of an E3 Ubiquitin-protein Ligase Activity,” Genes & Development, Vol. 13, pp.1822-1833 (1999)
17. Kane, C.J., Mallin, K., Ritchey, J., Cooperberg, M.R., and Carroll, P.R., “Renal Cell Cancer Stage Migration: Analysis of the National Cancer Data Base,” Cancer, Vol. 113 No.1, pp.78-83 (2008)
18. McLaughlin, J.K., and Lipworth, L., “Epidemiologic Aspects of Renal Cell Cancer,” Seminars in Oncology, Vol. 27, pp.115-123 (2000)
19. Calle, E.E., and Kaaks, R., “Overweight, Obesity and Cancer: Epidemiological Evidence and Proposed Mechanisms,” Nature Reviews Cancer, Vol. 4, No. 8, pp.579-591 (2004)
20. McLaughlin, J.K., Chow, W.H., Mandel, J.S., Mellemgaard, A., McCredie, M., Lindblad, P., Schlehofer, B., Pommer, W., Niwa, S., and Adami, H.O., “International Renal Cell Cancer Study: VIII. Role of Diuretics, Other Antihypertensive Medications and Hypertension,” International Journal of Cancer, Vol. 63, pp.216-221 (1995)
21. Robson, C.J., “Radical Nephrectomy for Renal Cell Carcinoma,” The Journal of Urology, Vol. 89, pp.37-42 (1963)
22. Uzzo, R.G., and Novick, A.C., “Nephron Sparing Surgery for Renal Tumors: Indications, Techniques and Outcomes,” The Journal of Urology, Vol. 166, No. 1, pp.6-18 (2001)
23. Abreu, S.C., and Gill, I.S., “Renal Cell Carcinoma: Modern Surgical Approach,” Current Opinion in Urology, Vol. 13, No. 6, pp.439-444 (2003)
24. Novick, A.C., “Kidney Cancer: Past, Present and Future,” Urologic Oncology, Vol. 25, pp.188-195 (2007)
25. Motzer, R.J., and Russo, P., “Systemic Therapy for Renal Cell Carcinoma,” The Journal of Urology, Vol. 163, pp.408-417 (2000)
26. Coppin, C., Prozsolt, F., Awa, A., Kumpf, J., Coldman, A., and Wilt, T., “Immunotherapy for Advanced Renal Cell Cancer,” The Cochrane Database of Systemic Reviews, Vol. 25, No. 1, pp.CD001425 (2005)
27. Motzer, R.J., Michaelson, M.D., Redman, B.G., Hudes, G.R., Wilding, G., Figlin, R.A., Ginsberg, M.S., Kim, S.T., Baum, C.M., DePrimo, S.E., Li, J.Z., Bello, C.L., Theuer, C.P., George, D.J., and Rini, B.I., “Activity of SU11248, a Multitargeted Inhibitor of Vascular Endothelial Growth Factor Receptor and Platelet-derived Growth Factor Receptor, in Patients with Metastatic Renal Cell Carcinoma,” Journal Clinical Oncology, Vol. 24, No. 1, pp.16-24 (2006)
28. Hudes, G., “Targeting mTOR in Renal Cell Carcinoma,” Cancer, Vol. 115, pp.2313-2320 (2009)
29. Goldar, S., Khaniani, M.S., Derakhshan, S.M., and Baradaran, B., “Molecular Mechanisms of Apoptosis and Roles in Cancer Development and Treatment,” Asian Pacific Journal of Cancer Prevention, Vol. 16, No. 6, pp.2129-2144 (2015)
30. Ashkenazi, A., “Targeting the Extrinsic Apoptotic Pathway in Cancer: Lessons Learned and Future Directions,” The Journal of Clinical Investigation, Vol. 125, No. 2, pp.487-489 (2015)
31. Patergnani, S., Missiroli, S., Marchi, S., and Giorgi, C., “Mitochondria-Associated Endoplasmic Reticulum Membranes Microenvironment: Targeting Autophagic and Apoptotic Pathways in Cancer Therapy,” Frontiers in Oncology, Vol. 27, No. 5, pp.173 (2015)
32. Hanahan, D., and Weinberg, R.A., “The Hallmarks of Cancer”, Cell, Vol. 100, No. 1, pp. 57-70 (2000)
33. Fidler, I.J., “The Pathogenesis of Cancer Metastasis: the 'Seed and Soil' Hypothesis Revisited,” Nature Reviews. Cancer, Vol. 3, No. 6, pp.453-458 (2003)
34. Chambers, A.F., Groom, A.C., and MacDonald, I.C., “Dissemination and Growth of Cancer Cells in Metastatic Sites,” Nature Reviews. Cancer, Vol. 2, No. 8, pp.563-572 (2002)
35. Ridley, A.J., Schwartz, M.A., Burridge, K., Firtel, R.A., Ginsberg, M.H., Borisy, G., Parsons, J.T., and Horwitz, A.R., “Cell Migration: Integrating Signals from Front to Back,” Science, Vol. 302, No. 5651, pp.1704-1709 (2003)
36. Derynck, R., Akhurst, R.J., and Balmain, A., “TGF-beta Signaling in Tumor Suppression and Cancer Progression,” Nature Genetics, Vol. 29, No.2, pp.117-129 (2001)
37. Parks, W.C., Wilson, C.L., and Lopez-Boado, Y.S., “Matrix Metalloproteinases as Modulators of Inflammation and Innate Immunity,” Nature Reviews. Immunology, Vol. 4, No. 8, pp.617-629 (2004)
38. Tan, W., Lu, J., Huang, M., Li, Y., Chen, M., Wu, G., Gong, J., Zhong, Z., Xu, Z., Dang, Y., Guo, J., Chen, X., and Wang, Y., “Anti-Cancer Natural Products Isolated from Chinese Medicinal Herbs,” Chinese Medicine, Vol. 6, No. 1, pp. 27-91 (2011)
39. Qiu, S., Sun, H., Zhang, A.H., Xu, H.Y., Yan, G.L., Han, Y., and Wang, X.J., “Natural Alkaloids: Basic Aspects, Biological Roles, and Future Perspectives,” Chinese Journal of Natural Medicines, Vol. 12, No. 6, pp.401-406 (2014)
40. Galassi, A., “The Next Generation: New Chemotherapy Agents for the 1990s” Seminas in Oncololy Nursing, Vol. 8, No. 2, pp.83-94 (1992)
41. Newman, D.J., and Cragg, G.M., “Natural Products as Sources of New Drugs over the 30 Years from 1981 to 2010,” Journal of Natural Products, Vol. 75, No. 3, pp.311-335 (2012)
42. Huang, T.T., Wu, S.P., Chong, K.Y., Ojcius, D.M., Ko, Y.F., Wu, Y.H., Wu, C.Y., Lu, C.C., Martel, J., Young, J.D., and Lai H.C., “The Medicinal Fungus Antrodia cinnamomea Suppresses Inflammation by Inhibiting the NLRP3 Inflammasome,” Journal of Ethnopharmacology, Vol. 155, No. 1, pp.154-164 (2014)
43. Kumar, K.J., Chu, F.H., Hsieh, H.W., Liao, J.W., Li, W.H., Lin, J.C., Shaw, J.F., and Wang, S.Y., “Antroquinonol from Ethanolic Extract of Mycelium of Antrodia cinnamomea Protects Hepatic Cells from Ethanol-Induced Oxidative Stress through Nrf-2 Activation,” Journal of Ethnopharmacology, Vol. 136, No. 1, pp.168-177 (2011)
44. Cheng, J.J., Huang, N.K., Chang, T.T., Wang, D.L., and Lu, M.K., ”Study for Anti-angiogenic Activities of Polysaccharides Isolated from Antrodia cinnamomea in Endothelial Cells,” Life Sciences, Vol. 76, No. 26, pp.3029-3042 (2005)
45. Yue, P.Y., Wong, Y.Y., Wong, K.Y., Tsoi, Y.K., and Leung, K.S., “Current Evidence for the Hepatoprotective Activities of the Medicinal Mushroom Antrodia cinnamomea,” Chinese Medicine, Vol. 8, No. 1, pp.21 (2013)
46. Lin, Y.W., Pan, J.H., Liu, R.H., Kuo, Y.H., Sheen, L.Y., and Chiang, B.H., “The 4-acetylantroquinonol B Isolated from Mycelium of Antrodia cinnamomea Inhibits Proliferation of Hepatoma Cells,” Journal of the Science of Food And Agriculture, Vol. 90, No. 10, pp.1739-1744 (2010)
47. Fa, K.N., Yang, C.M., Chen, P.C., Lee, Y.Y., Chyau, C.C., and Hu, M.L., “Anti-Metastatic Effects of Antrodan, the Antrodia cinnamomea Mycelia Glycoprotein, in Lung Carcinoma Cells,” Internaltional Journal of Biological Macromolecules, Vol. 74, pp.476-482 (2015)
48. Kumar, K.J., Vani, M.G., Chueh, P.J., Mau, J.L., and Wang, S.Y., “Antrodin C Inhibits Epithelial-to-Mesenchymal Transition and Metastasis of Breast Cancer Cells via Suppression of Smad2/3 and β-catenin Signaling Pathways,” PLoS One, Vol.10, No. 2, pp.e0117111 (2015)
49. Geethangili, M., Rao, Y.K., and Tzeng, Y.M., “Development and Validation of a HPLC-DAD Separation Method for Determination of Bioactive Antrocin in Medicinal Mushroom Antrodia camphorata,” International Journal of Applied Science and Engineering, Vol. 11, No. 2, pp.195-201 (2013)
50. Gokila Vani, M., Kumar, K.J., Liao, J.W., Chien, S.C., Mau, J.L., Chiang, S.S., Lin, C.C., Kuo, Y.H., and Wang S.Y., “Antcin C from Antrodia cinnamomea Protects Liver Cells Against Free Radical-Induced Oxidative Stress and Apoptosis in vitro and in vivo through Nrf2-Dependent Mechanism,” Evidence-Based Complementary and Alternative Medicine, Vol. 2013, pp.296082 (2013)
51. Lin, C.C., Chen, C.C., Kuo, Y.H., Kuo, J.T., Senthil Kumar, K.J., and Wang, S.Y. “2,3,5-Trimethoxy-4-cresol, an Anti-Metastatic Constituent from the Solid-State Cultured Mycelium of Antrodia cinnamomea and Its Mechanism,” Journal of Natural Medicines, Vol. 69, No. 4, pp.513-521 (2015)
52. Chen, Y.J., Chou, C.J., and Chang, T.T., “Compound MMH01 Possesses Toxicity against Human Leukemia and Pancreatic Cancer Cells,” Toxicology In Vitro, Vol. 23, No. 3, pp.418-424 (2009)
53. Chung, T.Y., Li, ,F.Y., Chang, C.I., Jinn, T.R., and Tzen, J.T., “Inhibition of Na(+)/K(+) -ATPase by Antcins, Unique Steroid-Like Compounds in Antrodia camphorate,” The American Journal of Chinese Medicine, Vol. 40, No. 5, pp.953-965 (2012)
54. Rao, Y.K., Wu, A.T., Geethangili, M., Huang, M.T., Chao, W.J., Wu, C.H., Deng, W.P., Yeh, C.T., and Tzeng, Y.M., “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,” Chemical Research in Toxicology, Vol. 24, No. 2, pp.238-245 (2010).
55. Yeh, C.T., Huang, W.C., Rao, Y.K., Ye, M., Lee, W.H., Wang, L.S., Tzeng, D.T., Wu, C.H., Shieh, Y.S., Huang, C.Y., Chen, Y.J., Hsiao, M., Wu, A.T., Yang, Z., and Tzeng, Y.M., “A Sesquiterpene Lactone Antrocin from Antrodia camphorata Negatively Modulates JAK2/STAT3 Signaling via microRNA let-7c and Induces Apoptosis in Lung Cancer Cells,” Carcinogenesis, pp.bgt255 (2013)
56. Kaufman, D.S., S.W., and Feldman, A.S., “Bladder Cancer,” Lancet, Vol. 374, pp. 239-249 (2009)
57. Arjonen, A., Kaukonen, R., and Ivaska, J., “Filopodia and Adhesion in Cancer Cell Motility,” Cell Adhesion & Migration, Vol. 5, No. 5, pp.421-430 (2011)
58. Zaidel-Bar, R., Ballestrem, C., Kam, Z., and Geiger, B., “Early Molecular Events in the Assembly of Matrix Adhesions at the Leading Edge of Migrating Cells,” Journal of Cell Science, Vol. 116, pp.4605-4613 (2003)
59. Fingleton, B., “Matrix Metalloproteinases: Roles in Cancer and Metastasis,” Frontiers in Bioscience: a Journal and Virtual Library, Vol. 11, pp.479-491 (2005)
60. Chen, Y.J., Wei, Y.Y., Chen, H.T., Fong, Y.C., Hsu, C.J., Tsai, C.H., Hsu, H.C., Liu, S.H., and Tang, C.H., “Osteopontin Increases Migration and MMP-9 Up‐Regulation via αvβ3 Integrin, FAK, ERK, and NF‐κB‐Dependent Pathway in Human Chondrosarcoma Cells,” Journal of Cellular Physiology, Vol. 221, No. 1, pp.98-108 (2009)
61. Webb, D.J., Donaisk, K., Whitmore, L.A., Thomas, S.M., Turner, C.E., Parsons, J.T., and Horwitz, A.F., “FAK-Src Signalling through Paxillin, ERK and MLCK Regulates Adhesion Disassembly,” Nature Cell Biology, Vol. 6, No. 2, pp.154-161 (2004)
62. Cohen, H.T., and McGovern, F.J., “Renal-Cell Carcinoma,” The New England Journal of Medicine, Vol. 353, No.23, pp.2477-2490 (2005)
63. Li, L., and Li, W., “Epithelial-Mesenchymal Transition in Human Cancer: Comprehensive Reprogramming of Metabolism, Epigenetics, and Differentiation,” Pharmacology & Therapeutics, Vol. 150, pp.33-46 (2015)
64. Leventhal, P.S., Shelden, E.A., Kim, B., and Feldman, E.L., “Tyrosine Phosphorylation of Paxillin and Focal Adhesion Kinase during Insulin-like Growth Factor-I-stimulated Lamellipodial Advance,” The Journal of Biological Chemistry, Vol. 272, pp.5214-5218 (1997)
65. Miyata, Y., Iwata, T., Ohba, K, Kanda, S., Nishikido, M., and Kanetake, H., “Expression of Matrix Metalloproteinase-7 on Cancer Cells and Tissue Endothelial Cells in Renal Cell Carcinoma: Prognostic Implications and Clinical Significance for Invasion and Metastasis,” Clinical Cancer Research, Vol. 12, No. 23, pp.6998-7003 (2006)
66. Ramankulov, A., Lein, M., Johannsen, M., Schrader, M., Miller, K., and Jung, K., “Plasma Matrix Metalloproteinase-7 as a Metastatic Marker and Survival Predictor in Patients with Renal Cell Carcinomas,” Cancer Science, Vol. 99, No. 6, pp.1188-1194 (2008)
67. Lee, J.M., Dedhar, S., Kalluri, R., and Thompson, E.W., “The Epithelial-Mesenchymal Transition: New Insights in Signaling, Development, and Disease,” The Journal of Cell Biology, Vol. 172, No. 7, pp.973-981 (2006)
68. Deramaud, T.B., Dujardin, D., Hamadi, A., Noulet, F., Kolli, K., De Mey, J., Takeda, K., and Rondé, P., “FAK Phosphorylation at Tyr-925 Regulates Cross-talk between Focal Adhesion Turnover and Cell Protrusion,” Molecular Biology of the Cell, Vol. 22, No. 7, pp.964-975 (2011)
69. Deramaudt, T.B., Dujardin, D., Noulet, F., Martin, S., Vauchelles, R., Takeda, K., and Rondé, P., “Altering FAK-Paxillin Interactions Reduces Adhesion, Migration and Invasion Processes,” PloS One, Vol. 9, No. 3, pp.e92059 (2014)
70. Mattila, P.K., and Lappalainen, P., “Filopodia: Molecular Architecture and Cellular Functions,” Nature Reviews. Molecular Cell Biology, Vol. 9, No. 6, pp.446-454 (2008)
71. Leventhal, P.S., Shelden, S.E., Kim, B., and Feldman, E.L., “Tyrosine Phosphorylation of Paxillin and Focal Adhesion Kinase during Insulin-like Growth Factor-I-stimulated Lamellipodial Advance,” The Journal of Biological Chemistry, Vol. 272, No. 8, pp.5214-5218 (1997)
72. Guan, J.L., “Role of Focal Adhesion Kinase in Integrin Signaling,” The Internaltional Journal of Biochemistry & Cell Biology, Vol. 28, No. 8-9, pp.1085-1096 (1997)
73. Hiscox, S., Barnfather, P., Hayes. E., Bramble, P., Christensen, J., Nicholson, R.I., and Barrett-Lee, P., “Inhibition of Focal Adhesion Kinase Suppresses the Adverse Phenotype of Endocrine-resistant Breast Cancer Cells and Improves Endocrine Response in Endocrine-sensitive Cells,” Breast Cancer Research and Treatment, Vol. 125, No. 3, pp.659-669 (2011)
74. Schlaepfer, D.D., Jones, K.C., and Hunter, T., “Multiple Grb2-mediated Integrin-stimulated Signaling Pathways to ERK2/mitogen-activated Protein Kinase: Summation of Both c-Src- and Focal Adhesion Kinase-initiated Tyrosine Phosphorylation Events,” Molecular and Cellular Biolology, Vol. 18, No. 5, pp.2571-2585 (1998)
75. Chaturvedi, L.S., Gayer, C.P., Marsh, H.M., and Basson, M.D., “Repetitive Deformation Activates Src-independent FAK-dependent ERK Motogenic Signals in Human Caco-2 Intestinal Epithelial Cells,” American Journal of Physiolology. Cell Physiology, Vol. 294, No.6, pp.1350-1361 (2008)
76. Schlaepfer, D.D., Hauck, C.R., and Sieg, D.J., “Signaling through Focal Adhesion Kinase,” Progress in Biophysics and Molecular Biology, Vol. 71, No. 3-4, pp.435-478 (1999)
77. Schlaepfer, D.D., Mitra, S.K., and Ilic, D., “Control of Motile and Invasive Cell Phenotypes by Focal Adhesion Kinase,” Biochimica et Biophysica Acta, Vol. 1692, No. 2-3, pp.77-102 (2004)
78. Mitra, S.K., Schlaepfer, D.D., “Integrin-regulated FAK-Src Signaling in Normal and Cancer Cells,” Current Opinion in Cell Biology, Vol. 18, No. 5, pp.516-523 (2006)
79. Bauvois, B., “New Facets of Matrix Metalloproteinases MMP-2 and MMP-9 as Cell Surface Transducers: Outside-in Signaling and Relationship to Tumor Progression,” Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, Vol. 1825, No. 1. pp.29-36 (2012)
80. Roy, R., Yang, J., and Moses, M.A., “Matrix Metalloproteinases as Novel Biomarker and Potential Therapeutic Targets in Human Cancer,” Journal of Clinical Oncology, Vol. 27, No. 31, pp.5287-5297 (2009)
81. Kumar, B., Koul, S., Petersen, J., Khandrika, L., Hwa, J.S., Meacham, R.B., Wilson, S., and Koul, H.K., “p38 Mitogen-Activated Protein Kinase–Driven MAPKAPK2 Regulates Invasion of Bladder Cancer by Modulation of MMP-2 and MMP-9 Activity,” Cancer Research, Vol. 70, No. 2 pp.832-841 (2010)
82. Gong, Y., Chippada-Venkata, U.D., and Oh, W.K., “Roles of Matrix Metalloproteinases and Their Natural Inhibitors in Prostate Cancer Progression,” Cancers, Vol. 6, No. 3, pp.1298-1327 (2014)
83.O'Donnell, A., Odrowaz, Z., and Sharrocks, A.D., “Immediate-early Gene Activation by the MAPK Pathways: What Do and Don't We Know?” Biochemical Society Transactions, Vol. 40, No. 1, pp.58-66 (2012)
84. Radisky, D.C., and LaBarge, M.A., “Epithelial-mesenchymal Transition and the Stem Cell Phenotype,” Cell Stem Cell, Vol. 2, No. 6, pp.511-512 (2008)
85. Nojima, D., Nakajima, K., Li, L.C., Franks, J., Ribeiro-Filho, L., Ishii, N., and Dahiya, R., “CpG Methylation of Promoter Region Inactivates E-cadherin Gene in Renal Cell Carcinoma,” Molecular Carcinogenesis, Vol. 32, No. 1, pp.19-27 (2001)
86. Katagiri, A., Watanabe, R., and Tomita, Y., “E-cadherin Expression in Renal Cell Cancer and Its Significance in Metastasis and Survival,” Britich Journal of Cancer, Vol. 71, No. 2, pp.376-9 (1995)
87. Ramankulov, A., Lein, M., Johannsen, M., Schrader, M., Miller, K., and Jung, K., “Plasma Matrix Metalloproteinase-7 as a Metastatic Marker and Survival Predictor in Patients with Renal Cell Carcinomas,” Cancer Science, Vol. 99, No. 6, pp.1188-1194 (2008)
88. Ii, M., Yamamoto, H., Adachi, Y., Maruyama, Y., and Shinomura, Y., “Role of Matrix Metalloproteinase-7 (Matrilysin) in Human Cancer Invasion, Apoptosis, Growth, and Angiogenesis,” Experimental Biology and Medicine (Maywood, N.J.), Vol. 231, No. 1, pp.20-27 (2006)
89. Lein, M., Jung, K., Laube, C., Hübner, T., Winkelmann, B., Stephan, C., Hauptmann, S., Rudolph, B., Schnorr, D., and Loening, S.A., “Matrix-metalloproteinases and Their Inhibitors in Plasma and Tumor Tissue of Patients with Renal Cell Carcinoma,” International Journal of Cancer, Vol. 85, No.6, pp.801-804 (2000)
90. Brew, K. and Nagase, H., “The Tissue Inhibitors of Metalloproteinases (TIMPs): an Ancient Family with Structural and Functional Diversity,” Biochimical Biophysiology Acta, Vol. 1803, No. 1, pp.55-71 (2010)
91. Han, H., Du, B., Pan, X., Liu, J., Zhao, Q., Lian, X., Qian, M., and Liu, M., “CADPE Inhibits PMA-stimulated Gastric Carcinoma Cell Invasion and Matrix Metalloproteinase-9 Expression by FAK/MEK/ERK-mediated AP-1 Activation,” Molecular Cancer Research, Vol. 8, No. 11, pp.1477-1488 (2010)
92. Verde, P., Casalino, L., Talotta, F., Yaniv, M., and Weitzman, J.B., “Deciphering AP-1 Function in Tumorigenesis: Fra-ternizing on Target Promoters,” Cell Cycle, Vol. 6, No. 21, pp.2633-1639 (2007)
93. Tu, Y.C., Huang, D.Y., Shiah, S.G., Wang, J.S., and Lin, W.W., “Regulation of c-Fos Gene Expression by NF-kappaB: a p65 Homodimer Binding Site in Mouse Embryonic Fibroblasts but not Human HEK293 Cells,” PLoS One, Vol. 8, No. 12, pp.e84062 (2013)
94. Chang, M.C., Chen, C.A., Chen, P.J., Chiang, Y.C., Chen, Y.L., Mao, T.L., Lin, H.W., Lin, Chiang, W.H., and Cheng, W.F., “Mesothelin Enhances Invasion of Ovarian Cancer by Inducing MMP-7 through MAPK/ERK and JNK Pathways,” The Biochemical Journal, Vol. 442, No. 2, pp.293-302 (2012)
95. Jia, Z.C., Wan, Y.L., Tang, J.Q., Dai, Y., Liu, Y.C., Wang, X., and Zhu, J., “Tissue Factor/Activated Factor VIIa Induces Matrix Metalloproteinase-7 Expression through Activation of c-Fos via ERK1/2 and p38 MAPK Signaling Pathways in Human Colon Cancer Cell,” International Journal of Colorectal Disease, Vol. 27, No. 4, pp. 437-445 (2012)
96. Ho, B.Y., Wu, Y.M., Chang, K.J., and Pan, T.M., “Dimerumic Acid Inhibits SW620 Cell Invasion by Attenuating H2O2-mediated MMP-7 Expression via JNK/C-Jun and ERK/C-Fos Activation in an AP-1-dependent Manner,” International Journal of Biololical Science, Vol. 7, No. 6, pp.869-880 (2011)
97. Cortez, D.M., Feldman, M.D., Mummidi, S., Valente, A.J., Steffensen, B., Vincenti, M., Barnes, J.L., and Chandrasekar, B., “IL-17 Stimulates MMP-1 Expression in Primary Human Cardiac Fibroblasts via p38 MAPK- and ERK1/2-dependent C/EBP-beta , NF-kappaB, and AP-1 Activation,” Americal Journal of Physiology. Heart and Circulatory Physiology, Vol. 293, No. 6, pp.H3356-3365 (2007)