臺灣博碩士論文加值系統

肺癌是國內外最常見的高死亡率癌症。由於癌細胞的轉移，造成病患預後狀況不
佳。癌轉移的過程中，癌細胞經由周邊血管或淋巴管移動至其他部位繼續分裂生
長，不僅會調控抑癌或致癌基因的表現，而且會分泌血管新生因子促進血管新生，加速癌轉移的發生。許多的藥理學家便嘗試著從保健草藥中尋求解答，以提供治療肺癌的新方向。然而，面對數以千萬計的保健藥材和未知的抗癌機轉，並沒有具專一性又具高效率的篩選藥物方法。根據實驗室先前的研究，我們已經發現 HLJ1 是一個新的抑癌基因且與非小細胞肺癌病人之預後存活率相關，可作為有潛力的藥物標靶。此外，血管內皮新生因子 (VEGF)與Src 會活化下游許多訊息傳遞路徑，導致血管新生與癌轉移，預期會是個有效的癌症治療標靶。在本研究中，我們將建立三個篩選藥物平台：第一與第二個平台為利用抑癌基因 (HLJ1)和血管新生因子 (VEGF) 啟動子之報導基因分析 (reporter gene assay) 模式來建立平台，進而篩選抑制肺癌細胞生長及調控血管新生之保健草藥並探討其作用機轉；第三個平台為發展電腦輔助藥物設計(CADD)平台來篩選有潛力的Src 抑制劑，這使我們容易於實驗室鑑定候選化合物，提高開發新藥物的可能性。利用報導基因之篩藥平台，我們從中草藥資料庫鑑定了數個草藥化合物可以增強HLJ1啟動子活性或減少VEGF 轉錄，進而抑制癌細胞的移動與侵襲。在這些草藥中，我們發現穿心蓮內酯 (andrographolide)確實可以顯著地促進HLJ1 蛋白質的表現並抑制體內外腫瘤生長，而且穿心蓮內酯也會經由活化JunB 來上調控HLJ1，進而調控AP-2α 結合至MMP-2 啟動子並抑制MMP-2 的表現。此外，由微陣列的分析結果指出穿心蓮內酯會影響細胞週期(cell cycle)、細胞凋亡(apoptosis)與細胞貼附相關訊息(adhesion-related biological signalling)，包括絲裂原活化蛋白激酶(mitogen-activated protein kinase)，貼附(focal adhesion)和緊密連接(tight junction)的途徑，這使得穿心蓮內酯具有抑制肺癌細胞侵入及生長的能力。另外，我們也發現毒毛旋花素(strophanthin)確實可以減少VEGF mRNA 的表現，並且有效地抑制肺癌細胞的侵襲、移動、非貼附性聚落形成、貼附性聚落形成的能力和活體內腫瘤的生長。而在血管生成(tube formation)試驗中顯示，毒毛旋花素也會抑制血管的生成。而且由即時定量聚合連鎖反應實驗證明，毒毛旋花素會抑制VEGF121 、VEGF165 以及VEGF189 的表現量。而第三個平台則是利用對接(docking)在SrcY418 的位置進行虛擬篩選，來尋找可以結合在Y418 位置的候選藥物。由實驗結果顯示有幾個化合物可以抑制Src 磷酸化，antihelminthic niclosamide 是其中一個化合物。實驗結果發現，niclosamide 確實會減少Src 磷酸化與肺癌細胞生存能力及誘導細胞凋亡，我們推測niclosamide 可能有臨床治療或預防肺癌進展的潛力。另外，我們進一步分析niclosamide 分子結構改變對於功效之影響。由實驗結果顯示在細胞生存能力中，niclosamide 其中一個衍生物(W3312)比niclosamide 更有效果。總之，niclosamide 可以抑制Src 活性與相關訊息途徑，並極具有臨床治療的潛力。綜合以上結果，我們建立了標的HLJ1 和VEGF 藥物篩選平台以及電腦輔助藥物設計平台，這些平台皆具有專一性並可快速地篩選新的抗癌化合物。利用這些平台，我們確定穿心蓮內酯、毒毛旋花素與niclosamide是很有潛力的新抗癌藥物，可以抑制非小細胞肺癌的腫瘤生長和侵襲。

High mortality lung cancer is the most frequent cause of cancer deaths worldwide,including Taiwan. Because of cancer cell metastasis, cancer patients have poor prognosis. The process of cancer metastasis, cancer cells via peripheral vascular or lymphatic move to other parts of the division and growth. The process not only regulate tumor suppressor gene or oncogene expression, cancer cells also secrete angiogenic factors to promote angiogenesis and accelerate cancer metastasis. A lot of pharmacologists try to find the answers from the traditional Chinese herb medicines. However, there is no specific and high-throughput way to screen thousands of these “health-care” herbs to unknown anti-cancer pathway. According to our previous studies, we have identified a novel suppressor gene (HLJ1) which might relate to patients’ survival rate in NSCLC and can be used as a potential drug target. The vascular endothelial angiogenic factors (VEGF) plays essential roles in the activation of many downstream signalling pathways, promotion of angiogenesis and cancer metastasis and is expected to be a potent target for cancer therapy. In this study, we establish three platforms of drug screening. The first and second platforms were the HLJ1 and VEGF-targeting drug-screening platforms which analyze the HLJ1 and VEGF promoter activities by reporter gene assay. Utilizing these platforms, we can screen herbal medicines with lung cancer cell growth inhibition and angiogenesis regulation and investigate their mechanisms. Third platform was computer-aided drug design (CADD) method, we used CADD to develope novel c-Src inhibitors. By CADD, it is easy to identify candidate compounds for biological validation, and increase the successful rate of new drug development. Utilizing drug screening platforms of reporter gene, we identified several herbal compounds from a Chinese herbal library with the capacity to enhance HLJ1 promoter activity or inhibit VEGF transcription and thereby inhibited cancer cell migration and invasion. Among the herbal drugs identified the andrographolide most significantly induced HLJ1 expression and suppressed tumorigenesis both in vitro and in vivo. The andrographolide upregulated HLJ1 via JunB activation, which modulates AP-2α binding at the MMP-2 promoter and represses the expression of MMP-2. Microarray transcriptomic analysis was performed to comprehensively depict the andrographolide-regulated signalling pathways. We showed that andrographolide can affect genes that are dominantly involved in the cell cycle, apoptosis and adhesion-related biological signalling, including mitogen-activated protein kinase, focal adhesion and tight junction pathways, indicating the diverse effects of andrographolide on anticancer invasion and proliferation. In addition, we also found that strophanthin could decrease VEGF mRNA expression and inhibit cancer cell invasion, migration, anchorage-independent and -dependent growth and tumor growth. The tube formation assay was revealed that strophanthin can suppress angiogenesis. Furthermore, the expression of VEGF121, VEGF165 and VEGF189 of VEGF isoforms were repressed by strophanthin. The third platform, computational virtual screening of Src inhibitor by docking on Y418 site, is performed to find the candidate drugs which could bind on Src Y418 site. The results showed that several compounds can suppress Src phosphorylation, especially antihelminthic niclosamide. We demonstrated that niclosamide could reduce src phorsphorylation and lung cancer cell viability, and induce apoptosis, suggesting that niclosamide may have the potential for the clinical treatment or prevention of lung cancer progression in humans. Moreover, we also analyzed the effect of structural changes in the niclosamide molecule on its ability. The result showed that one of the niclosamide derivatives (W3312) was more effective than niclosamide in cell viability. In summary, niclosamide can suppress Src activity and related signaling pathway and make great potential for the clinical treatment. In conclusion, the HLJ1-targeting, VEGF-targeting drug-screening platforms and CADD are useful for screening of novel anticancer compounds. Using these platforms, we identified andrographolide, strophanthin and niclosamide potentially as promising new anticancer agents that could suppress tumor growth and invasion in NSCLC.

目錄
中文摘要....................................................i
英文摘要................................................….iii
目錄.....................................................….v
表目錄...................................................viii
圖表目錄.............................……....................ix
緒論
一、肺癌....................................................1
二、癌轉移..................................................3
三、中草藥與治療癌症..........................................4
四、篩選平台與方式............................................5
五、熱休克蛋白...............................................7
六、Human Liver DnaJ-like protein (HLJ1)..................10
七、血管新生與血管新生因子....................................11
八、血管內皮新生因子 (VEGF)..................................12
九、Src 酪氨酸激酶 (tyrosine kinase) .......................15
十、穿心蓮內酯 (andrographolide) ...........................19
十一、毒毛旋花素 (ouabain) ..................................20
十二、Niclosamide..........................................21
研究目的...................................................23
實驗材料與方法
一、 細胞株的選用…...........................................24
二、 細胞培養與分盤..........................................24
三、選擇中草藥與藥劑準備......................................25
四、質體的製備..............................................26
五、 細胞轉染作用............................................29
六、冷光報導基因分析.........................................30
七、細胞total RNA 萃取......................................31
八、 cDNA 的合成與即時定量RT-PCR 分析.........................32
九、細胞蛋白質之抽取 (protein extration) 與定量 (protein amount assay)...33
十、西方墨點法..............................................34
十一、噻唑藍試驗 ( MTT assay )...............................35
十二、細胞聚落形成分析 (Colony formation).....................36
十三、細胞增生分析 ( Proliferation assay )...................36
十四、流式細胞分析技術 (Flow cytometery)......................37
十五、細胞遷移分析 (Migration assay)…........................38
十六、細胞基質侵襲力分析 (Matrigel invasion assay)............38
十七、管柱形成實驗 (Tube formation assay)....................39
十八、膠質酶譜分析(gelatin zymography).......................39
十九、染色質免疫沈澱分析(Chromatin immunoprecipitation assay ；
ChIP )....................................................40
二十、腫瘤新生試驗 (Tumorigenesis assay).....................41
二十一、統計分析.............................................41
實驗結果
一、測定中草藥對CL1-5 細胞的毒性..............................42
二、利用報導基因分析平台發現有22 種中草藥在CL1-5 細胞中可以促使HLJ1啟動子冷光酵素活性增..........................................42
三、20 種中草藥對肺腺癌細胞CL1-5 中的HLJ1 蛋白質表現有影響…......42
四、確認CHM-10、CHM-37、CHM-45、CHM-76 和CHM-82 可以在CL1-5細胞中增加HLJ1 的表現////........................................43
五、中草藥增加HLJ1 的表現進而影響肺腺癌細胞CL1-5 侵入的能力........43
六、中草藥增加HLJ1 的表現進而影響肺腺癌細胞CL1-5 遷移的能力........43
七、穿心蓮內酯可以促進HLJ1 之表現..............................44
八、穿心蓮內酯經由JunB 蛋白上調控HLJ1 之表現....................44
九、穿心蓮內酯可抑制癌細胞的增生與腫瘤生長.......................45
十、分析穿心蓮內酯誘導的基因轉錄和基因網絡.......................46
十一、穿心蓮內酯會抑制肺癌細胞的細胞週期過程......................46
十二、穿心蓮內酯會誘導肺癌細胞的凋亡............................47
十三、 HLJ1 對於穿心蓮內酯影響抗侵襲所扮演的角色.................47
十四、穿心蓮內酯對於細胞外基質與基質降解蛋白的影響.................48
十五、穿心蓮內酯抑制肺癌腫瘤形成的假設模型.......................48
十六、利用報導基因分析平台初步篩選出抑制VEGF 活性之候選藥物........48
十七、利用報導基因分析平台篩選出抑制肺腺癌細胞株中VEGF 活性之候選藥物.......................................................49
十八、利用報導基因分析平台篩選出之八種候選藥物均可抑制肺腺癌細胞株CL1-5 中VEGF mRNA 表現量........................................49
十九、利用MTT assay 測定毒毛旋花素對肺腺癌CL1-5 細胞的毒性.......49
二十、毒毛旋花素對於肺腺癌細胞CL1-5 增生能力的影響...............50
二十一、毒毛旋花素對於肺腺癌細胞CL1-5 貼附性生長的影響.............50
二十二、毒毛旋花素對於肺腺癌細胞CL1-5 非貼附性生長的影響...........50
二十三、毒毛旋花素)對人類臍靜脈內皮細胞形成管狀構造的影響...........50
二十四、毒毛旋花素對於肺腺癌細胞CL1-5 侵襲能力的影響..............51
二十五、毒毛旋花素對於肺腺癌細胞CL1-5 移動能力的影響..............51
二十六、觀察毒毛旋花素對裸鼠的皮下腫瘤生長之影響..................51
二十七、觀察毒毛旋花素對VEGF 亞型，VEGF121、VEGF165 以及VEGF189
mRNA 表現量的影響..........................................52
二十八、觀察毒毛旋花素對VEGF 下游蛋白AKT 磷酸化的影響…............52
二十九、毒毛旋花素抑制肺癌腫瘤形成的假設模型......................52
三十、利用計算機輔助藥物設計進行藥物虛擬篩選......................52
三十一、利用西方墨點法觀察不同肺腺癌細胞珠磷酸化src 之表現與驗證虛擬對接之結果.....................................................53
三十二、抗寄生蟲藥劑niclosamide 對肺腺癌細胞的影響...............53
三十三、Niclosamide 對肺癌細胞中src 相關基因和途徑之表現有時間依賴性
與濃度依賴性的影響...........................................54
三十四、在肺腺癌細胞中，niclosamide 會導致自體吞噬的產生..........54
三十五、Niclosamide 經由活化AIF 因子使細胞走向非依賴caspase 之細胞凋
亡途徑.....................................................55
三十六、AIF 在niclosamide 抑制肺癌細胞株A549 之角色............56
三十七、肺癌細胞株A549 被niclosamide 影響之基因與訊息途徑表現.... 56
三十八、Niclosamide 衍生物會抑制src 相關基因和途徑..............56
三十九、Niclosamide 衍生物會抑制肺癌細胞株的增生能力.............57
四十、Niclosamide 確定影響與可能影響之訊息途徑..................57
討論......................................................58
結論......................................................66
參考文獻...................................................67

參考文獻
1. Siegel R, Naishadham D, Jemal A (2012) Cancer statistics, 2012. CA Cancer J Clin 62: 10-29.
2. Dela Cruz CS, Tanoue LT, Matthay RA (2011) Lung cancer: epidemiology, etiology, and prevention. Clin Chest Med 32: 605-644.
3. McErlean A, Ginsberg MS (2011) Epidemiology of lung cancer. Seminars in Roentgenol 46: 173-177.
4. Bilello KS, Murin S, Matthay RA (2002) Epidemiology, etiology, and prevention of lung cancer. Clin Chest Med 23: 1-25.
5. Sun S, Schiller JH, Gazdar AF (2007) Lung cancer in never smokers--a different disease. Nat Rev Cancer 7: 778-790.
6. Thun MJ, Hannan LM, Adams-Campbell LL, Boffetta P, Buring JE, et al. (2008) Lung cancer occurrence in never-smokers: an analysis of 13 cohorts and 22 cancer registry studies. PLoS Med 5: e185.
7. Lan Q, Hsiung CA, Matsuo K, Hong YC, Seow A, et al. (2012) Genome-wide association analysis identifies new lung cancer susceptibility loci in never-smoking women in Asia. Nature Genet 44: 1330-1335.
8. Collins LG, Haines C, Perkel R, Enck RE (2007) Lung Cancer: Diagnosis and Management. Am Family Physician.
9. Hoffman PC, Mauer AM, Vokes EE (2000) Lung cancer. The Lancet.
10. Schuchert MJ, Luketich JD (2003) Solitary sites of metastatic disease in non-small cell lung cancer. Curr Treat Options Oncol 4: 65-79.
11. Pao W, Girard N (2011) New driver mutations in non-small-cell lung cancer. Lancet Oncol 12: 175-180.
12. Rosell R, Carcereny E, Gervais R, Vergnenegre A, Massuti B, et al. (2012) Erlotinib versus standard chemotherapy as first-line treatment for European patients with advanced EGFR mutation-positive non-small-cell lung cancer (EURTAC): a multicentre, open-label, randomised phase 3 trial. Lancet Oncol 13: 239-246.
13. Kobayashi S, Boggon TJ, Dayaram T, Janne PA, Kocher O, et al. (2005) EGFR mutation and resistance of non-small-cell lung cancer to gefitinib. N Engl J Med 352: 786-792.
14. Pao W (2012) New approaches to targeted therapy in lung cancer. Proc Am Thorac Soc 9: 72-73.
15. Chaffer CL, Weinberg RA (2011) A perspective on cancer cell metastasis. Science 331: 1559-1564.
16. Geiger TR, Peeper DS (2009) Metastasis mechanisms. Biochim Biophys Acta 1796: 293-308.
17. Nguyen DX, Massague J (2007) Genetic determinants of cancer metastasis. Nat Rev Genet 8: 341-352.
18. Huber MA, Kraut N, Beug H (2005) Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 17: 548-558.
19. Stupack DG, Teitz T, Potter MD, Mikolon D, Houghton PJ, et al. (2006) Potentiation of neuroblastoma metastasis by loss of caspase-8. Nature 439: 95-99.
20. Minn AJ, Gupta GP, Siegel PM, Bos PD, Shu W, et al. (2005) Genes that mediate breast cancer metastasis to lung. Nature 436: 518-524.
21. Gupta GP, Nguyen DX, Chiang AC, Bos PD, Kim JY, et al. (2007) Mediators of vascular remodelling co-opted for sequential steps in lung metastasis. Nature 446: 765-770.
22. NS C (2003) Metastatic bladder cancer: role of chemotherapy and new agents. EAU Update Ser 1: 108-117.
23. Sadava D, Ahn J, Zhan M, Pang ML, Ding J, et al. (2002) Effects of four Chinese herbal extracts on drug-sensitive and multidrug-resistant small-cell lung carcinoma cells. Cancer Chemother Pharmacol 49: 261-266.
24. KC. H (1999) The pharmacology of Chinese herbs, 2nd edition. Boca Raton7 CRC Press.
25. Schwartsmann G, Ratain MJ, Cragg GM, Wong JE, Saijo N, et al. (2002) Anticancer drug discovery and development throughout the world. J Clin Oncol 20: 47S-59S.
26. Hsiao WL, Liu L (2010) The role of traditional Chinese herbal medicines in cancer therapy--from TCM theory to mechanistic insights. Planta Med 76: 1118-1131.
27. Pao W (2012) New approaches to targeted therapy in lung cancer. Annals of The A T S 9: 72-73.
28. Dorai T, Aggarwal BB (2004) Role of chemopreventive agents in cancer therapy. Cancer Lett 215: 129-140.
29. Notte A, Ninane N, Arnould T, Michiels C (2013) Hypoxia counteracts taxol-induced apoptosis in MDA-MB-231 breast cancer cells: role of autophagy and JNK activation. Cell Death Dis 4: e638.
30. Cai XZ, Huang WY, Qiao Y, Du SY, Chen Y, et al. (2013) Inhibitory effects of curcumin on gastric cancer cells: a proteomic study of molecular targets. Phytomedicine 20: 495-505.
31. Du GJ, Zhang Z, Wen XD, Yu C, Calway T, et al. (2012) Epigallocatechin Gallate (EGCG) is the most effective cancer chemopreventive polyphenol in green tea. Nutrients 4: 1679-1691.
32. Tang FY, Cho HJ, Pai MH, Chen YH (2009) Concomitant supplementation of lycopene and eicosapentaenoic acid inhibits the proliferation of human colon cancer cells. J Nutr Biochem 20: 426-434.
33. Ohnuki S, Oka S, Nogami S, Ohya Y (2010) High-content, image-based screening for drug targets in yeast. PLoS One 5: e10177.
34. Saenz A, Lopez de Munain A (2008) DNA arrays: a general overview and specific applications. Med Clin (Barc) 130: 504-509.
35. Johnson PH, Walker RP, Jones SW, Stephens K, Meurer J, et al. (2002) Multiplex gene expression analysis for high-throughput drug discovery: screening and analysis of compounds affecting genes overexpressed in cancer cells. Mol Cancer Ther 1: 1293-1304.
36. Guo L, Lian JH, Ji W, Hu WR, Wu GL, et al. (2006) Establishment of a cell-based drug screening system for identifying selective down-regulators of mPGES-1. Inflamm Res 55: 114-118.
37. Meng XY, Zhang HX, Mezei M, Cui M (2011) Molecular docking: a powerful approach for structure-based drug discovery. Curr Comput Aided Drug Des 7: 146-157.
38. Sack U, Walther W, Scudiero D, Selby M, Kobelt D, et al. (2011) Novel effect of antihelminthic Niclosamide on S100A4-mediated metastatic progression in colon cancer. J Natl Cancer Inst 103: 1018-1036.
39. Nithya V, Halami PM (2012) Novel whole-cell Reporter Assay for Stress-Based Classification of Antibacterial Compounds Produced by Locally Isolated Bacillus spp. Indian J Microbiol 52: 180-184.
40. Fuentes S, Crim RL, Beeler J, Teng MN, Golding H, et al. (2013) Development of a simple, rapid, sensitive, high-throughput luciferase reporter based microneutralization test for measurement of virus neutralizing antibodies following respiratory syncytial virus vaccination and infection. Vaccine.
41. Liu AM, New DC, Lo RK, Wong YH (2009) Reporter gene assays. Methods Mol Biol 486: 109-123.
42. Kitchen DB, Decornez H, Furr JR, Bajorath J (2004) Docking and scoring in virtual screening for drug discovery: methods and applications. Nat Rev Drug Discov 3: 935-949.
43. Nandi S, Bagchi MC (2009) 3D-QSAR and molecular docking studies of 4-anilinoquinazoline derivatives: a rational approach to anticancer drug design. Mol Divers 14: 27-38.
44. Okimoto N, Futatsugi N, Fuji H, Suenaga A, Morimoto G, et al. (2009) High-performance drug discovery: computational screening by combining docking and molecular dynamics simulations. PLoS Comput Biol 5: e1000528.
45. Chen GL, Wang LH, Wang J, Chen K, Zhao M, et al. (2013) Discovery of a small molecular compound simultaneously targeting RXR and HADC: Design, synthesis, molecular docking and bioassay. Bioorg Med Chem Lett 23: 3891-3895.
46. Bello M, Martinez-Archundia M, Correa-Basurto J (2013) Automated docking for novel drug discovery. Expert Opin Drug Discov.
47. Young JC, Agashe VR, Siegers K, Hartl FU (2004) Pathways of chaperone-mediated protein folding in the cytosol. Nature Reviews Mol Cell Biol 5: 781-791.
48. Jego G, Hazoume A, Seigneuric R, Garrido C (2010) Targeting heat shock proteins in cancer. Cancer Letters.
49. Lanneau D, Wettstein G, Bonniaud P, Garrido C (2010) Heat shock proteins: cell protection through protein through Protein Triage. Sci World J 10: 1543-1552.
50. Kampinga HH, Hageman J, Vos MJ, Kubota H, Tanguay RM, et al. (2009) Guidelines for the nomenclature of the human heat shock proteins. Cell Stress Chaperones 14: 105-111.
51. Mitra A, Shevde LA, Samant RS (2009) Multi-faceted role of HSP40 in cancer. Clin Exp Metastasis 26: 559-567.
52. Jego G, Hazoume A, Seigneuric R, Garrido C (2010) Targeting heat shock proteins in cancer. Cancer Lett 332: 275-285.
53. Dragovic Z, Broadley SA, Shomura Y, Bracher A, Hartl FU (2006) Molecular chaperones of the Hsp110 family act as nucleotide exchange factors of Hsp70s. EMBO J 25: 2519-2528.
54. Murphy ME (2013) The HSP70 family and cancer. Carcinogenesis 34: 1181-1188.
55. Kelley WL, Georgopoulos C (1992) Chaperones and Protein Folding. Cur Opinion in Cell Biol 4: 984-991.
56. Shimamura T, Li D, Ji H, Haringsma HJ, Liniker E, et al. (2008) Hsp90 inhibition suppresses mutant EGFR-T790M signaling and overcomes kinase inhibitor resistance. Cancer Res 68: 5827-5838.
57. Huang Q, Ye J, Chen W, Wang L, Lin W, et al. (2010) Heat shock protein 27 is over-expressed in tumor tissues and increased in sera of patients with gastric adenocarcinoma. Clin Chem Lab Med 48: 263-269.
58. Kocsis J, Madaras B, Toth EK, Fust G, Prohaszka Z (2010) Serum level of soluble 70-kD heat shock protein is associated with high mortality in patients with colorectal cancer without distant metastasis. Cell Stress Chaperones 15: 143-151.
59. Schorey JS, Bhatnagar S (2008) Exosome function: from tumor immunology to pathogen biology. Traffic 9: 871-881.
60. Vega VL, Rodriguez-Silva M, Frey T, Gehrmann M, Diaz JC, et al. (2008) Hsp70 translocates into the plasma membrane after stress and is released into the extracellular environment in a membrane-associated form that activates macrophages. J Immunol 180: 4299-4307.
61. Gibert B, Eckel B, Fasquelle L, Moulin M, Bouhallier F, et al. (2012) Knock down of heat shock protein 27 (HspB1) induces degradation of several putative client proteins. PLoS One 7: e29719.
62. Kamada M, So A, Muramaki M, Rocchi P, Beraldi E, et al. (2007) Hsp27 knockdown using nucleotide-based therapies inhibit tumor growth and enhance chemotherapy in human bladder cancer cells. Mol Cancer Ther 6: 299-308.
63. Mitra A, Shevde LA, Samant RS (2009) Multi-faceted role of HSP40 in cancer. Clin Exp Metastasis 26: 559-567.
64. Hessenkemper W, Baniahmad A (2013) Targeting heat shock proteins in prostate cancer. Curr Med Chem.
65. Kim LS, Kim JH (2011) Heat shock protein as molecular targets for breast cancer therapeutics. J Breast Cancer 14: 167-174.
66. Ciocca DR, Arrigo AP, Calderwood SK (2013) Heat shock proteins and heat shock factor 1 in carcinogenesis and tumor development: an update. Arch Toxicol 87: 19-48.
67. Yeh CH, Tseng R, Hannah A, Estrov Z, Estey E, et al. (2010) Clinical correlation of circulating heat shock protein 70 in acute leukemia. Leuk Res 34: 605-609.
68. Nakamura H, Minegishi H (2013) HSP60 as a drug target. Curr Pharm Des 19: 441-451.
69. Nakajima M, Kato H, Miyazaki T, Fukuchi M, Masuda N, et al. (2011) Prognostic significance of heat shock protein 110 expression and T lymphocyte infiltration in esophageal cancer. Hepatogastroenterology 58: 1555-1560.
70. Slaby O, Sobkova K, Svoboda M, Garajova I, Fabian P, et al. (2009) Significant overexpression of Hsp110 gene during colorectal cancer progression. Oncol Rep 21: 1235-1241.
71. Cherneva RV, Georgiev OB, Petrova DS, Trifonova NL, Stamenova M, et al. (2012) The role of small heat-shock protein alphaB-crystalline (HspB5) in COPD pathogenesis. Int J Chron Obstruct Pulmon Dis 7: 633-640.
72. Sterrenberg JN, Blatch GL, Edkins AL (2011) Human DNAJ in cancer and stem cells. Cancer Letters 312: 129-142.
73. Chen JJ, Peck K, Hong TM, Yang SC, Sher YP, et al. (2001) Global analysis of gene expression in invasion by a lung cancer model. Cancer Res 61: 5223-5230.
74. Tsai MF, Wang CC, Chang GC, Chen CY, Chen HY, et al. (2006) A new tumor suppressor DnaJ-like heat shock protein, HLJ1, and survival of patients with non-small-cell lung carcinoma. J Natl Cancer Inst 98: 825-838.
75. Ohtsuka K, Hata M (2000) Mammalian HSP40/DNAJ homologs: cloning of novel cDNAs and a proposal for their classification and nomenclature. Cell Stress Chaperones 5: 98-112.
76. Hamajima F, Hasegawa T, Nakashima I, Isobe K (2002) Genomic cloning and promoter analysis of the GAHSP40 gene. J Cell Biochem 84: 401-407.
77. Chang TP, Yu SL, Lin SY, Hsiao YJ, Chang GC, et al. (2010) Tumor suppressor HLJ1 binds and functionally alters nucleophosmin via activating enhancer binding protein 2alpha complex formation. Cancer Res 70: 1656-1667.
78. Lin SY, Hsueh CM, Yu SL, Su CC, Shum WY, et al. (2010) HLJ1 is a novel caspase-3 substrate and its expression enhances UV-induced apoptosis in non-small cell lung carcinoma. Nucleic Acids Res 38: 6148-6158.
79. Chen CH, Lin H, Chuang SM, Lin SY, Chen JJ (2010) Acidic stress facilitates tyrosine phosphorylation of HLJ1 to associate with actin cytoskeleton in lung cancer cells. Exp Cell Res 316: 2910-2921.
80. Chen HW, Lee JY, Huang JY, Wang CC, Chen WJ, et al. (2008) Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1. Cancer Res 68: 7428-7438.
81. Wang CC, Lin SY, Lai YH, Liu YJ, Hsu YL, et al. (2012) Dimethyl sulfoxide promotes the multiple functions of the tumor suppressor HLJ1 through activator protein-1 activation in NSCLC cells. PLoS One 7: e33772.
82. Wang CC, Tsai MF, Hong TM, Chang GC, Chen CY, et al. (2005) The transcriptional factor YY1 upregulates the novel invasion suppressor HLJ1 expression and inhibits cancer cell invasion. Oncogene 24: 4081-4093.
83. Wang CC, Tsai MF, Dai TH, Hong TM, Chan WK, et al. (2007) Synergistic activation of the tumor suppressor, HLJ1, by the transcription factors YY1 and activator protein 1. Cancer Res 67: 4816-4826.
84. Verde P, Casalino L, Talotta F, Yaniv M, Weitzman JB (2007) Deciphering AP-1 function in tumorigenesis: fra-ternizing on target promoters. Cell Cycle 6: 2633-2639.
85. Eferl R, Wagner EF (2003) AP-1: a double-edged sword in tumorigenesis. Nat Rev Cancer 3: 859-868.
86. Mburu YK, Egloff AM, Walker WH, Wang L, Seethala RR, et al. (2012) Chemokine receptor 7 (CCR7) gene expression is regulated by NF-kappaB and activator protein 1 (AP1) in metastatic squamous cell carcinoma of head and neck (SCCHN). J Biol Chem 287: 3581-3590.
87. Ibrahim EE, Babaei-Jadidi R, Saadeddin A, Spencer-Dene B, Hossaini S, et al. (2012) Embryonic NANOG activity defines colorectal cancer stem cells and modulates through AP1- and TCF-dependent mechanisms. Stem Cells 30: 2076-2087.
88. Coultas L, Chawengsaksophak K, Rossant J (2005) Endothelial cells and VEGF in vascular development. Nature 438: 937-945.
89. Naumov GN, Folkman J, Straume O, Akslen LA (2008) Tumor-vascular interactions and tumor dormancy. APMIS 116: 569-585.
90. Almog N, Ma L, Raychowdhury R, Schwager C, Erber R, et al. (2009) Transcriptional switch of dormant tumors to fast-growing angiogenic phenotype. Cancer Res 69: 836-844.
91. Kerbel RS (2008) Tumor angiogenesis. N Engl J Med 358: 2039-2049.
92. Adams RH, Alitalo K (2007) Molecular regulation of angiogenesis and lymphangiogenesis. Nat Rev Mol Cell Biol 8: 464-478.
93. van Hinsbergh VW, Koolwijk P (2008) Endothelial sprouting and angiogenesis: matrix metalloproteinases in the lead. Cardiovasc Res 78: 203-212.
94. Vincenti V, Cassano C, Rocchi M, Persico G (1996) Assignment of the vascular endothelial growth factor gene to human chromosome 6p21.3. Circulation 93: 1493-1495.
95. Cursiefen C, Chen L, Borges LP, Jackson D, Cao J, et al. (2004) VEGF-A stimulates lymphangiogenesis and hemangiogenesis in inflammatory neovascularization via macrophage recruitment. J Clin Invest 113: 1040-1050.
96. Ferrara N, Gerber HP, LeCouter J (2003) The biology of VEGF and its receptors. Nat Med 9: 669-676.
97. Klettner A, Roider J (2009) Treating age-related macular degeneration - interaction of VEGF-antagonists with their target. Mini Rev Med Chem 9: 1127-1135.
98. Matsumoto T, Claesson-Welsh L (2001) VEGF receptor signal transduction. Sci STKE 2001: re21.
99. Ahmed Z, Bicknell R (2009) Angiogenic signalling pathways. Methods Mol Biol 467: 3-24.
100. Los M, Roodhart JM, Voest EE (2007) Target practice: lessons from phase III trials with bevacizumab and vatalanib in the treatment of advanced colorectal cancer. Oncologist 12: 443-450.
101. Rosenfeld PJ, Heier JS, Hantsbarger G, Shams N (2006) Tolerability and efficacy of multiple escalating doses of ranibizumab (Lucentis) for neovascular age-related macular degeneration. Ophthalmology 113: 623 e621.
102. Higa GM, Abraham J (2007) Lapatinib in the treatment of breast cancer. Expert Rev Anticancer Ther 7: 1183-1192.
103. Roskoski R, Jr. (2007) Sunitinib: a VEGF and PDGF receptor protein kinase and angiogenesis inhibitor. Biochem Biophys Res Commun 356: 323-328.
104. Wilhelm SM, Adnane L, Newell P, Villanueva A, Llovet JM, et al. (2008) Preclinical overview of sorafenib, a multikinase inhibitor that targets both Raf and VEGF and PDGF receptor tyrosine kinase signaling. Mol Cancer Ther 7: 3129-3140.
105. Kelly RJ, Rixe O (2009) Axitinib--a selective inhibitor of the vascular endothelial growth factor (VEGF) receptor. Target Oncol 4: 297-305.
106. Podar K, Tonon G, Sattler M, Tai YT, Legouill S, et al. (2006) The small-molecule VEGF receptor inhibitor pazopanib (GW786034B) targets both tumor and endothelial cells in multiple myeloma. Proc Natl Acad Sci U S A 103: 19478-19483.
107. Sloan B, Scheinfeld NS (2008) Pazopanib, a VEGF receptor tyrosine kinase inhibitor for cancer therapy. Curr Opin Investig Drugs 9: 1324-1335.
108. Hurwitz H, Saini S (2006) Bevacizumab in the treatment of metastatic colorectal cancer: safety profile and management of adverse events. Semin Oncol 33: S26-34.
109. Sandler A, Gray R, Perry MC, Brahmer J, Schiller JH, et al. (2006) Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer. N Engl J Med 355: 2542-2550.
110. Chang JH, Garg NK, Lunde E, Han KY, Jain S, et al. (2012) Corneal neovascularization: an anti-VEGF therapy review. Surv Ophthalmol 57: 415-429.
111. Kamba T, McDonald DM (2007) Mechanisms of adverse effects of anti-VEGF therapy for cancer. Br J Cancer 96: 1788-1795.
112. Morabito A, De Maio E, Di Maio M, Normanno N, Perrone F (2006) Tyrosine kinase inhibitors of vascular endothelial growth factor receptors in clinical trials: current status and future directions. Oncologist 11: 753-764.
113. Pieramici DJ, Rabena MD (2008) Anti-VEGF therapy: comparison of current and future agents. Eye (Lond) 22: 1330-1336.
114. Holash J, Davis S, Papadopoulos N, Croll SD, Ho L, et al. (2002) VEGF-Trap: a VEGF blocker with potent antitumor effects. Proc Natl Acad Sci U S A 99: 11393-11398.
115. Stewart MW, Rosenfeld PJ (2008) Predicted biological activity of intravitreal VEGF Trap. Br J Ophthalmol 92: 667-668.
116. Oliveira HB, Sakimoto T, Javier JA, Azar DT, Wiegand SJ, et al. (2010) VEGF Trap(R1R2) suppresses experimental corneal angiogenesis. Eur J Ophthalmol 20: 48-54.
117. Martin GS (2001) The hunting of the Src. Nat Rev Mol Cell Biol 2: 467-475.
118. Thomas SM, Brugge JS (1997) Cellular functions regulated by Src family kinases. Annu Rev Cell Dev Biol 13: 513-609.
119. Le XF, Bast RC, Jr. (2011) Src family kinases and paclitaxel sensitivity. Cancer Biol Ther 12: 260-269.
120. Wheeler DL, Iida M, Dunn EF (2009) The role of Src in solid tumors. Oncologist 14: 667-678.
121. Brown MT, Cooper JA (1996) Regulation, substrates and functions of src. Biochim Biophys Acta 1287: 121-149.
122. Zheng XM, Resnick RJ, Shalloway D (2000) A phosphotyrosine displacement mechanism for activation of Src by PTPalpha. EMBO J 19: 964-978.
123. Roskoski R, Jr. (2004) Src protein-tyrosine kinase structure and regulation. Biochem Biophys Res Commun 324: 1155-1164.
124. Aleshin A, Finn RS (2010) SRC: a century of science brought to the clinic. Neoplasia 12: 599-607.
125. Summy JM, Gallick GE (2003) Src family kinases in tumor progression and metastasis. Cancer Metastasis Rev 22: 337-358.
126. Elsberger B, Fullerton R, Zino S, Jordan F, Mitchell TJ, et al. (2010) Breast cancer patients'' clinical outcome measures are associated with Src kinase family member expression. Br J Cancer 103: 899-909.
127. Tan M, Li P, Klos KS, Lu J, Lan KH, et al. (2005) ErbB2 promotes Src synthesis and stability: novel mechanisms of Src activation that confer breast cancer metastasis. Cancer Res 65: 1858-1867.
128. Wiener JR, Windham TC, Estrella VC, Parikh NU, Thall PF, et al. (2003) Activated SRC protein tyrosine kinase is overexpressed in late-stage human ovarian cancers. Gynecol Oncol 88: 73-79.
129. Rosano L, Cianfrocca R, Masi S, Spinella F, Di Castro V, et al. (2009) Beta-arrestin links endothelin A receptor to beta-catenin signaling to induce ovarian cancer cell invasion and metastasis. Proc Natl Acad Sci U S A 106: 2806-2811.
130. Zhang Q, Thomas SM, Xi S, Smithgall TE, Siegfried JM, et al. (2004) SRC family kinases mediate epidermal growth factor receptor ligand cleavage, proliferation, and invasion of head and neck cancer cells. Cancer Res 64: 6166-6173.
131. van Oijen MG, Rijksen G, ten Broek FW, Slootweg PJ (1998) Overexpression of c-Src in areas of hyperproliferation in head and neck cancer, premalignant lesions and benign mucosal disorders. J Oral Pathol Med 27: 147-152.
132. Masaki T, Igarashi K, Tokuda M, Yukimasa S, Han F, et al. (2003) pp60c-src activation in lung adenocarcinoma. Eur J Cancer 39: 1447-1455.
133. Zhang J, Kalyankrishna S, Wislez M, Thilaganathan N, Saigal B, et al. (2007) SRC-family kinases are activated in non-small cell lung cancer and promote the survival of epidermal growth factor receptor-dependent cell lines. Am J Pathol 170: 366-376.
134. Lee D, Gautschi O (2006) Clinical development of SRC tyrosine kinase inhibitors in lung cancer. Clin Lung Cancer 7: 381-384.
135. Giaccone G, Zucali PA (2008) Src as a potential therapeutic target in non-small-cell lung cancer. Ann Oncol 19: 1219-1223.
136. Yu-Ming Chang LB, Joy Yang, Hsing-Jien Kung and Christopher Evans (2006) Survey of Src activity and Src-related growth and migration in prostate cancer lines. Proc Amer Assoc Cancer Res 47: 2505a.
137. Song L, Turkson J, Karras JG, Jove R, Haura EB (2003) Activation of Stat3 by receptor tyrosine kinases and cytokines regulates survival in human non-small cell carcinoma cells. Oncogene 22: 4150-4165.
138. Laird AD, Li G, Moss KG, Blake RA, Broome MA, et al. (2003) Src family kinase activity is required for signal tranducer and activator of transcription 3 and focal adhesion kinase phosphorylation and vascular endothelial growth factor signaling in vivo and for anchorage-dependent and -independent growth of human tumor cells. Mol Cancer Ther 2: 461-469.
139. Guarino M, Rubino B, Ballabio G (2007) The role of epithelial-mesenchymal transition in cancer pathology. Pathology 39: 305-318.
140. Wei L, Yang Y, Zhang X, Yu Q (2004) Altered regulation of Src upon cell detachment protects human lung adenocarcinoma cells from anoikis. Oncogene 23: 9052-9061.
141. Guilhot F, Apperley J, Kim DW, Bullorsky EO, Baccarani M, et al. (2007) Dasatinib induces significant hematologic and cytogenetic responses in patients with imatinib-resistant or -intolerant chronic myeloid leukemia in accelerated phase. Blood 109: 4143-4150.
142. Song L, Morris M, Bagui T, Lee FY, Jove R, et al. (2006) Dasatinib (BMS-354825) selectively induces apoptosis in lung cancer cells dependent on epidermal growth factor receptor signaling for survival. Cancer Res 66: 5542-5548.
143. F. R. Luo FRL, Y. Barrett, P. Ji, P. Holly, E. McCann, P. Rhyne, E. Clarke, K. He, E. Bleichardt and M. Blackwood-Chirchir (2006) Dasatinib (BMS-354825) pharmacokinetics correlate with pSRC pharmacodynamics in phase I studies of patients with cancer (CA180002, CA180003). J of Clin Oncology 24: 18s (Abstr 3046).
144. W. A. Messersmith SK, B. A. Hewes, C. M. Zacharchuk, R. Abbas, P. Martins, E. Dowling, A. Volkert, E. Martin and A. I. Daud (2007) Bosutinib (SKI-606), a dual Src/Abl tyrosine kinase inhibitor: preliminary results from a phase 1 study in patients with advanced malignant solid tumours. J of Clin Oncology 25: 18S (Abstr 3552).
145. O. Gautschi PP, C. P. Evans, J. C. Yang, W. S. Holland, R. J. Bold, S. Virudachalam, P. N. Lara, D. R. Gandara and P. H. Gumerlock (2006) Preclinical evaluation of the dual specific Src/Abl kinase inhibitor AZD0530 in lung cancer. J of Clin Oncology 24: 18s(Abstr 13108).
146. Hiscox S, Morgan L, Green TP, Barrow D, Gee J, et al. (2006) Elevated Src activity promotes cellular invasion and motility in tamoxifen resistant breast cancer cells. Breast Cancer Res Treat 97: 263-274.
147. Hennequin LF, Allen J, Breed J, Curwen J, Fennell M, et al. (2006) N-(5-chloro-1,3-benzodioxol-4-yl)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5- (tetrahydro-2H-pyran-4-yloxy)quinazolin-4-amine, a novel, highly selective, orally available, dual-specific c-Src/Abl kinase inhibitor. J Med Chem 49: 6465-6488.
148. J. Cooper MMM, J. Curtright, A. Ricart, A. Mita, C. Takimoto, A. Tolcher, A. Dowlati, S. Flick and K. P. Papadopoulos (2006) A phase I study examining weekly dosing and pharmacokinetics (PK) of a novel spectrum selective kinase inhibitor, XL999, in patients (pts) with advanced solid malignancies (ASM). J of Clin Oncology 24: 18s (Abstr 13024).
149. Antonarakis ES, Heath EI, Posadas EM, Yu EY, Harrison MR, et al. (2013) A phase 2 study of KX2-391, an oral inhibitor of Src kinase and tubulin polymerization, in men with bone-metastatic castration-resistant prostate cancer. Cancer Chemother Pharmacol 71: 883-892.
150. Zhao J, Yang G, Liu H, Wang D, Song X, et al. (2002) Determination of andrographolide, deoxyandrographolide and neoandrographolide in the Chinese herb Andrographis paniculata by micellar electrokinetic capillary chromatography. Phytochem Anal 13: 222-227.
151. Reddy VL, Reddy SM, Ravikanth V, Krishnaiah P, Goud TV, et al. (2005) A new bis-andrographolide ether from Andrographis paniculata nees and evaluation of anti-HIV activity. Nat Prod Res 19: 223-230.
152. Shen YC, Chen CF, Chiou WF (2002) Andrographolide prevents oxygen radical production by human neutrophils: possible mechanism(s) involved in its anti-inflammatory effect. Br J Pharmacol 135: 399-406.
153. Jada SR, Hamzah AS, Lajis NH, Saad MS, Stevens MF, et al. (2006) Semisynthesis and cytotoxic activities of andrographolide analogues. J Enzyme Inhib Med Chem 21: 145-155.
154. Satyanarayana C, Deevi DS, Rajagopalan R, Srinivas N, Rajagopal S (2004) DRF 3188 a novel semi-synthetic analog of andrographolide: cellular response to MCF 7 breast cancer cells. BMC Cancer 4: 26.
155. Shi MD, Lin HH, Lee YC, Chao JK, Lin RA, et al. (2008) Inhibition of cell-cycle progression in human colorectal carcinoma Lovo cells by andrographolide. Chem Biol Interact 174: 201-210.
156. Zhou J, Zhang S, Ong CN, Shen HM (2006) Critical role of pro-apoptotic Bcl-2 family members in andrographolide-induced apoptosis in human cancer cells. Biochem Pharmacol 72: 132-144.
157. Sheeja K, Kuttan G (2007) Activation of cytotoxic T lymphocyte responses and attenuation of tumor growth in vivo by Andrographis paniculata extract and andrographolide. Immunopharmacol Immunotoxicol 29: 81-93.
158. Cheung HY, Cheung SH, Li J, Cheung CS, Lai WP, et al. (2005) Andrographolide isolated from Andrographis paniculata induces cell cycle arrest and mitochondrial-mediated apoptosis in human leukemic HL-60 cells. Planta Med 71: 1106-1111.
159. Shen KK, Liu TY, Xu C, Ji LL, Wang ZT (2009) Andrographolide inhibits hepatoma cells growth and affects the expression of cell cycle related proteins. Yao Xue Xue Bao 44: 973-979.
160. Kim TG, Hwi KK, Hung CS (2005) Morphological and biochemical changes of andrographolide-induced cell death in human prostatic adenocarcinoma PC-3 cells. In Vivo 19: 551-557.
161. Yang L, Wu D, Luo K, Wu S, Wu P (2009) Andrographolide enhances 5-fluorouracil-induced apoptosis via caspase-8-dependent mitochondrial pathway involving p53 participation in hepatocellular carcinoma (SMMC-7721) cells. Cancer Lett 276: 180-188.
162. Tsai HR, Yang LM, Tsai WJ, Chiou WF (2004) Andrographolide acts through inhibition of ERK1/2 and Akt phosphorylation to suppress chemotactic migration. Eur J Pharmacol 498: 45-52.
163. Shi MD, Lin HH, Chiang TA, Tsai LY, Tsai SM, et al. (2009) Andrographolide could inhibit human colorectal carcinoma Lovo cells migration and invasion via down-regulation of MMP-7 expression. Chem Biol Interact 180: 344-352.
164. Lee YC, Lin HH, Hsu CH, Wang CJ, Chiang TA, et al. (2010) Inhibitory effects of andrographolide on migration and invasion in human non-small cell lung cancer A549 cells via down-regulation of PI3K/Akt signaling pathway. Eur J Pharmacol 632: 23-32.
165. Lin HH, Tsai CW, Chou FP, Wang CJ, Hsuan SW, et al. (2011) Andrographolide down-regulates hypoxia-inducible factor-1alpha in human non-small cell lung cancer A549 cells. Toxicol Appl Pharmacol 250: 336-345.
166. Lingrel JB (2010) The physiological significance of the cardiotonic steroid/ouabain-binding site of the Na,K-ATPase. Annu Rev Physiol 72: 395-412.
167. Chanvorachote P, Pongrakhananon V (2012) Ouabain downregulates Mcl-1 and sensitizes lung cancer cells to TRAIL-induced apoptosis. Am J Physiol Cell Physiol 304: C263-272.
168. Xu ZW, Wang FM, Gao MJ, Chen XY, Shan NN, et al. (2011) Cardiotonic steroids attenuate ERK phosphorylation and generate cell cycle arrest to block human hepatoma cell growth. J Steroid Biochem Mol Biol 125: 181-191.
169. Prassas I, Diamandis EP (2008) Novel therapeutic applications of cardiac glycosides. Nat Rev Drug Discov 7: 926-935.
170. Zhang H, Qian DZ, Tan YS, Lee K, Gao P, et al. (2008) Digoxin and other cardiac glycosides inhibit HIF-1alpha synthesis and block tumor growth. Proc Natl Acad Sci U S A 105: 19579-19586.
171. Xu ZW, Wang FM, Gao MJ, Chen XY, Hu WL, et al. (2010) Targeting the Na(+)/K(+)-ATPase alpha1 subunit of hepatoma HepG2 cell line to induce apoptosis and cell cycle arresting. Biol Pharm Bull 33: 743-751.
172. Wang Y, Qiu Q, Shen JJ, Li DD, Jiang XJ, et al. (2012) Cardiac glycosides induce autophagy in human non-small cell lung cancer cells through regulation of dual signaling pathways. Int J Biochem Cell Biol 44: 1813-1824.
173. Yang P, Menter DG, Cartwright C, Chan D, Dixon S, et al. (2009) Oleandrin-mediated inhibition of human tumor cell proliferation: importance of Na,K-ATPase alpha subunits as drug targets. Mol Cancer Ther 8: 2319-2328.
174. Mijatovic T, De Neve N, Gailly P, Mathieu V, Haibe-Kains B, et al. (2008) Nucleolus and c-Myc: potential targets of cardenolide-mediated antitumor activity. Mol Cancer Ther 7: 1285-1296.
175. Mijatovic T, Roland I, Van Quaquebeke E, Nilsson B, Mathieu A, et al. (2007) The alpha1 subunit of the sodium pump could represent a novel target to combat non-small cell lung cancers. J Pathol 212: 170-179.
176. Lefranc F, Kiss R (2008) The sodium pump alpha1 subunit as a potential target to combat apoptosis-resistant glioblastomas. Neoplasia 10: 198-206.
177. Winnicka K, Bielawski K, Bielawska A, Surazynski A (2008) Antiproliferative activity of derivatives of ouabain, digoxin and proscillaridin A in human MCF-7 and MDA-MB-231 breast cancer cells. Biol Pharm Bull 31: 1131-1140.
178. Raghavendra PB, Sreenivasan Y, Ramesh GT, Manna SK (2007) Cardiac glycoside induces cell death via FasL by activating calcineurin and NF-AT, but apoptosis initially proceeds through activation of caspases. Apoptosis 12: 307-318.
179. Craig P, Ito A (2007) Intestinal cestodes. Curr Opin Infect Dis 20: 524-532.
180. Merschjohann K, Steverding D (2008) In vitro trypanocidal activity of the anti-helminthic drug niclosamide. Exp Parasitol 118: 637-640.
181. Osada T, Chen M, Yang XY, Spasojevic I, Vandeusen JB, et al. (2011) Antihelminth compound niclosamide downregulates Wnt signaling and elicits antitumor responses in tumors with activating APC mutations. Cancer Res 71: 4172-4182.
182. Jin Y, Lu Z, Ding K, Li J, Du X, et al. (2010) Antineoplastic mechanisms of niclosamide in acute myelogenous leukemia stem cells: inactivation of the NF-kappaB pathway and generation of reactive oxygen species. Cancer Res 70: 2516-2527.
183. Yo YT, Lin YW, Wang YC, Balch C, Huang RL, et al. (2012) Growth inhibition of ovarian tumor-initiating cells by niclosamide. Mol Cancer Ther 11: 1703-1712.
184. Bafico A, Liu G, Goldin L, Harris V, Aaronson SA (2004) An autocrine mechanism for constitutive Wnt pathway activation in human cancer cells. Cancer Cell 6: 497-506.
185. Li Y, Lu W, He X, Schwartz AL, Bu G (2004) LRP6 expression promotes cancer cell proliferation and tumorigenesis by altering beta-catenin subcellular distribution. Oncogene 23: 9129-9135.
186. Lu W, Lin C, Roberts MJ, Waud WR, Piazza GA, et al. (2011) Niclosamide suppresses cancer cell growth by inducing Wnt co-receptor LRP6 degradation and inhibiting the Wnt/beta-catenin pathway. PLoS One 6: e29290.
187. Wang AM, Ku HH, Liang YC, Chen YC, Hwu YM, et al. (2009) The autonomous notch signal pathway is activated by baicalin and baicalein but is suppressed by niclosamide in K562 cells. J Cell Biochem 106: 682-692.
188. Balgi AD, Fonseca BD, Donohue E, Tsang TC, Lajoie P, et al. (2009) Screen for chemical modulators of autophagy reveals novel therapeutic inhibitors of mTORC1 signaling. PLoS One 4: e7124.
189. Conway TF, Jr., Sabel MS, Sugano M, Frelinger JG, Egilmez NK, et al. (2000) Growth of human tumor xenografts in SCID mice quantified using an immunoassay for tumor marker protein in serum. J Immunol Methods 233: 57-65.
190. Zheng C (2006) Image Analysis and Cytometry in Three-Dimensional Digital Reconstruction of Porcine Native Aortic Valve Leaflets.
191. Kumar RA, Sridevi K, Kumar NV, Nanduri S, Rajagopal S (2004) Anticancer and immunostimulatory compounds from Andrographis paniculata. J Ethnopharmacol 92: 291-295.
192. Liu CC, Lin CC, Chen WS, Chen HY, Chang PC, et al. (2006) CRSD: a comprehensive web server for composite regulatory signature discovery. Nucleic Acids Res 34: W571-577.
193. Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2: 161-174.
194. Chang TP, Yu SL, Lin SY, Hsiao YJ, Chang GC, et al. (2010) Tumor suppressor HLJ1 binds and functionally alters nucleophosmin via activating enhancer binding protein 2alpha complex formation. Cancer Res 70: 1656-1667.
195. Maycotte P, Thorburn A (2011) Autophagy and cancer therapy. Cancer Biol Ther 11: 127-137.
196. Miramar MD, Costantini P, Ravagnan L, Saraiva LM, Haouzi D, et al. (2001) NADH oxidase activity of mitochondrial apoptosis-inducing factor. J Biol Chem 276: 16391-16398.
197. Cande C, Vahsen N, Garrido C, Kroemer G (2004) Apoptosis-inducing factor (AIF): caspase-independent after all. Cell Death Differ 11: 591-595.
198. Sheeja K, Guruvayoorappan C, Kuttan G (2007) Antiangiogenic activity of Andrographis paniculata extract and andrographolide. Int Immunopharmacol 7: 211-221.
199. Mishra SK, Sangwan, N.S., Sangwan, R.S. (2007) Andrographis paniculata (Kalmegh): a review. Pharmacog Rev 1: 283-298.
200. Pholphana N, Rangkadilok N, Thongnest S, Ruchirawat S, Ruchirawat M, et al. (2004) Determination and variation of three active diterpenoids in Andrographis paniculata (Burm.f.) Nees. Phytochem Anal 15: 365-371.
201. Chandrasekaran CV, Thiyagarajan P, Sundarajan K, Goudar KS, Deepak M, et al. (2009) Evaluation of the genotoxic potential and acute oral toxicity of standardized extract of Andrographis paniculata (KalmCold). Food Chem Toxicol 47: 1892-1902.
202. Poolsup N, Suthisisang C, Prathanturarug S, Asawamekin A, Chanchareon U (2004) Andrographis paniculata in the symptomatic treatment of uncomplicated upper respiratory tract infection: systematic review of randomized controlled trials. J Clin Pharm Ther 29: 37-45.
203. Rajagopal S, Kumar RA, Deevi DS, Satyanarayana C, Rajagopalan R (2003) Andrographolide, a potential cancer therapeutic agent isolated from Andrographis paniculata. J Exp Ther Oncol 3: 147-158.
204. Suzuki M, Hosaka Y, Matsushima H, Goto T, Kitamura T, et al. (1999) Butyrolactone I induces cyclin B1 and causes G2/M arrest and skipping of mitosis in human prostate cell lines. Cancer Lett 138: 121-130.
205. Zhang J, Li Y, Guo L, Cao R, Zhao P, et al. (2009) DH166, a beta-carboline derivative, inhibits the kinase activity of PLK1. Cancer Biol Ther 8: 2374-2383.
206. Warren CD, Brady DM, Johnston RC, Hanna JS, Hardwick KG, et al. (2002) Distinct chromosome segregation roles for spindle checkpoint proteins. Mol Biol Cell 13: 3029-3041.
207. Nilsson J, Yekezare M, Minshull J, Pines J (2008) The APC/C maintains the spindle assembly checkpoint by targeting Cdc20 for destruction. Nat Cell Biol 10: 1411-1420.
208. Musacchio A, Salmon ED (2007) The spindle-assembly checkpoint in space and time. Nat Rev Mol Cell Biol 8: 379-393.
209. Mojtahedi A, Salehi R, Navabakbar F, Tamizifar H, Tavakkoli H, et al. (2007) Evaluation of apoptosis induction using PARP cleavage on gastric adenocarcinoma and fibroblast cell lines by different strains of Helicobacter pylori. Pak J Biol Sci 10: 4097-4102.
210. Fumarola C, Guidotti GG (2004) Stress-induced apoptosis: toward a symmetry with receptor-mediated cell death. Apoptosis 9: 77-82.
211. Jean D, Gershenwald JE, Huang S, Luca M, Hudson MJ, et al. (1998) Loss of AP-2 results in up-regulation of MCAM/MUC18 and an increase in tumor growth and metastasis of human melanoma cells. J Biol Chem 273: 16501-16508.
212. Gast RE, Konig S, Rose K, Ferenz KB, Krieglstein J (2011) Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells. BMC Biochem 12: 28.
213. Wu CJ, Jan JT, Chen CM, Hsieh HP, Hwang DR, et al. (2004) Inhibition of severe acute respiratory syndrome coronavirus replication by niclosamide. Antimicrob Agents Chemother 48: 2693-2696.
214. Zhu PJ, Hobson JP, Southall N, Qiu C, Thomas CJ, et al. (2009) Quantitative high-throughput screening identifies inhibitors of anthrax-induced cell death. Bioorg Med Chem 17: 5139-5145.
215. Chen M, Wang J, Lu J, Bond MC, Ren XR, et al. (2009) The anti-helminthic niclosamide inhibits Wnt/Frizzled1 signaling. Biochemistry 48: 10267-10274.
216. Fonseca BD, Diering GH, Bidinosti MA, Dalal K, Alain T, et al. (2012) Structure-activity analysis of niclosamide reveals potential role for cytoplasmic pH in control of mammalian target of rapamycin complex 1 (mTORC1) signaling. J Biol Chem 287: 17530-17545.
217. Pospisilik JA, Knauf C, Joza N, Benit P, Orthofer M, et al. (2007) Targeted deletion of AIF decreases mitochondrial oxidative phosphorylation and protects from obesity and diabetes. Cell 131: 476-491.
218. Weinbach EC, Garbus J (1969) Mechanism of action of reagents that uncouple oxidative phosphorylation. Nature 221: 1016-1018.