肺癌在台灣所造成的死亡人數逐步增加，已經是主要的十大癌症死亡原因之一，尤其增加的又以女性及年輕男性患者增加最多，所以，不得不加以防治。當肺癌細胞轉移（metastasis）到身體的別處時就很難以外科方式加以治療，只能加以使用化學治療或放射性療法，但其副作用很大，病人的預後也很差。因此，須要積極開發可以有效抑制癌細胞轉移的藥物，以幫忙癌症的治療。所以，針對此一議題，本實驗發現了新的抗轉移藥物DMHMPC及DE-DMPC，這二者可以抑制人類肺癌細胞A549的轉移及侵入（invasion）作用。從420種小分子化學復合物中篩選出，發現利用Boyden chamber分析A549細胞株轉移實驗，結果發現DMHMPC和DE-DMPC會抑制A549細胞株的轉移及侵犯活性。MTT分析可以知道不會對A549細胞株具有毒性。因癌細胞的移動性很強，所以利用Wound healing 來分析，發現DMHMPC和DE-DMPC也都可抑制A549細胞株的移動性。本實驗希望此二者藥物並不會影響細胞的生長，所以進一步利用生長曲線分析跟colony formation assay分析其抑制細胞生長之能力，結果發現DMHMP及DE-DMPC並不會影響A549細胞株的生長。因此，這些種種的實驗結果建議DMHMPC及DE-DMPC為一具潛力之新的可以抑制癌細胞轉移的小分子化合物，它將可有效的幫忙癌症的治療，也將是在癌症治療或預防癌症上一個新契機。

Lung cancer is the leading cause of death in Taiwan cancer patients. The cancer cells are capable of spreading by metastasis and invasion. When tumors spread to distant parts of the body, it is difficult to treat by surgery. In this study, we discovered the novel anti-invasion drugs DMHMPC and DE-DMPC that can inhibit the metastasis and invasion of human lung cancer cell A549. From screening of a chemical library of 420 small molecule compounds using Boyden chamber and MTT assay, the compound DMHMPC and DE-DMPC showed the inhibitory activity toward the invasiveness but not proliferation in human lung cancer cell A549. The results of wound healing assay also showed that DMHMPC and DE-DMPC inhibited the motility of A549 in a dose dependent manner. Moreover, the results of growth curve assay and colony formation assay also showed that DMHMPC and DE-DMPC are not effect proliferation in A549. Together, these results suggested that DMHMPC and DE-DMPC could be the potential drugs to inhibit the invasion of cancer cells that may lead to the new avenue of the cancer prevention therapy.

中文摘要-----------------------------------------------------4
英文摘要-----------------------------------------------------5
縮寫-----------------------------------------------------------6
導論-----------------------------------------------------------7
肺癌-----------------------------------------------------------7
肺癌症狀-----------------------------------------------------7
診斷方式-----------------------------------------------------8
肺癌的分類--------------------------------------------------8
肺癌的分期--------------------------------------------------9
轉移-----------------------------------------------------------9
肺癌的治療--------------------------------------------------10
材料與方法--------------------------------------------------11
細胞培養-----------------------------------------------------11
細胞生長活性分析（MTT assay） --------------------11
Boyden Chamber：Migration活性試驗--------------12
Boyden Chamber：Invasion活性試驗---------------12
Wound Healing 移動性試驗-----------------------------13
Growth curve 生長曲線分析-----------------------------14
Colony formation 生長菌落分析-----------------------14
結果-----------------------------------------------------------15
討論-----------------------------------------------------------20
表與圖：
表1. 各細胞株對藥物的生長活性分析-----------------24
圖1. A549細胞株的migration分析結果---------------25
圖2. A549細胞株的invasion分析結果----------------27
圖3. DMHMPC在A549細胞株的移動性試驗---------29
圖4. A549細胞株在含有DMHMPC的環境下其生長曲線-------------------31
圖5. A549細胞株在含有DMHMPC的環境下其生長菌落情形--------------32
圖6. DE-DMPC在A549細胞株的移動性試驗----------------------------------35
圖7. A549細胞株在含有DE-DMPC的環境下其生長曲線----------------37
圖8. A549細胞株在含有DE-DMPC的環境下其生長菌落情形--------------38
參考文獻---------------------------------------------------------41
附錄：
附錄一 Saframycin的結構式--------------------------------49
附錄二DMHMPC的結構式----------------------------------50
附錄三DE-DMPC的結構式----------------------------------51

1. 行政院衛生署衛生統計資訊網歷年癌症死亡原因，表13（1-6）。
2. 財團法人台灣癌症基金會會訊，第34期，2008。
3. Jasti S. Rao, Christopher Gondi, Chandramu Chetty, Subramanyam Chittivelu, Pushpa A. Joseph, and Sajani S. Lakka. Inhibition of invasion, angiogenesis, tumor growth, and metastasis by adenovirus-mediated transfer of antisense uPAR and MMP-9 in non-small cell lung cancer cells. Mol Cancer Ther, 4:1399-1408, 2005.
4. Lewis J. Kleinsmith. Principles of Cancer Biology. 260, 2006.
5. MacKay AR, Corbitt RH, Hartzler JL, Thorgeirsson UP. Basement membrane type IV collagen degradation: evidence for the involvement of a proteolytic cascade independent of metalloproteinases. Cancer Res, 50:5997–6001, 1990.
6. Liotta LA, Stetler-Stevenson WG. Tumor invasion and metastasis: an imbalance of positive and negative regulation. Cancer Res, 51:5054–9s, 1991.
7. Vassalli JD. The urokinase receptor. Fibrinolysis, 8:172-181, 1994.
8. Minoru Shiraga, Seiji Yano, Akihiko Yamamoto, Hirohisa Ogawa, Hisatsugu Goto, Toyokazu Miki, Keisuke Miki, Helong Zhang, and Saburo Sone. Organ Heterogeneity of Host-derived Matrix Metalloproteinase Expression and Its Involvement in Multiple-Organ Metastasis by Lung Cancer Cell Lines. Cancer Res, 62:5967-5973, 2002.
9. Kanayama H., Yokota K., Kurokawa Y., Murakami Y., Nishitani M., Kagawa S. Prognostic values of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expression in bladder cancer. Cancer, 82:1359-1363, 1998.
10. Sakata K., Shigemasa K., Nagai N., Ohama K. Expression of matrix metalloproteinases (MMP-2, MMP-9, MT1-MMP) and their inhibitors (TIMP-1, TIMP-2) in common epithelial tumors of the ovary. Int J Oncol, 17:673-681, 2000.
11. Yu-An Chang, Tsung-Hsien Sun, Michael Y. Chiang, Pei-Jung Lu, Yi-Ting Huang, Li-Ching Liang and Chi Wi Ong. Short synthesis of piperizinohydroisoquinoline ring by selective Pictet-Spengler cyclization and evaluation of antitumor activity. Tetrahedron, 63: 8781-8787, 2007.
12. Chi-Wi Ong, Yu-An Chang, Jing-Yun Wu and Chien-Chung Cheng. Novel design of a pentacyclic scaffold as structural mimic of saframycin A. Tetrahedron, 59:8245-8249, 2003.
13. Cheng-Chi Chang, Jin-Yuan Shih, Yung-Ming Jeng, Jen-Liang Su, Been-Zen Lin, Szu-Ta Chen, Yat-Pang Chau, Pan-Chyr Yang, Min-Liang Kuo. Connective Tissue Growth Factor and Its Role in Lung Adenocarcinoma Invasion and Metastasis. J Natl Cancer Inst, 96:364-375, 2004.
14. Arundhati Ghosh, Xinning Wang, Eric Klein, and Warren D.W. Heston. Novel Role of Prostate-Specific Membrane Antigen in Suppressing Prostate Cancer Invasiveness. Cancer Res, 65:727-731, 2005.
15. Elaine M.Langenfeld, Steve E.Calvano , Fadi Abou-Nukta, Stephen F.Lowry, Peter Amenta and John Langenfeld. The mature bone morphogenetic protein-2 is aberrantly expressed in non-small cell lung carcinomas and stimulates tumor growth of A549 cells. Carcinogenesis, 24:1445-1454, 2003.
16. Christof R. Hauck, David J. Sieg, Datsun A. Hsia, Joseph C. Loftus, William A. Gaarde, Brett P. Monia, and David D. Schlaepfer. Inhibition of Focal Adhesion Kinase Expression or Activity Disrupts Epidermal Growth Factor-stimulated Signaling Promoting the Migration of Invasive Human Carcinoma Cells. Cancer Res, 61: 7079-7090, 2001.
17. Eric S. White, Scott R. B. Strom, Noel L. Wys, and Douglas A. Arenberg. Non-Small Cell Lung Cancer Cells Induce Monocytes to Increase Expression of Angiogenic Activity. J Immunol, 166: 7549–7555, 2001.
18. Ashraf Dallol, Angelo Agathanggelou, Stella Tommasi, Gerd P. Pfeifer, Eamonn R. Maher, and Farida Latif. Involvement of the RASSF1A Tumor Suppressor Gene in Controlling Cell Migration. Cancer Res, 65(17):7653-7659, 2005.
19. Svitlana M. Kurinna, Chun Chui Tsao, Alina Felicia Nica, Tilahun Jiffar, and Peter P. Ruvolo. Ceramide Promotes Apoptosis in Lung Cancer-Derived A549 Cells by a Mechanism Involving c-Jun NH2-Terminal Kinase. Cancer Res, 64:7852-7856, 2004.
20. Jang-Seong Kim, Ji-Hoon Chang, Hyun-Kyung Yu, Jin-Hyung Ahn, Jung-Sun Yum, Suk-Keun Lee, Kyung-Hwan Jung, Doo-Hong Park, Yeup Yoon, Si-Myung Byun, and Soo-Il Chung. Inhibition of Angiogenesis and Angiogenesis-dependent Tumor Growth by the Cryptic Kringle Fragments of Human Apolipoprotein. J Biol Chem, 278:29000-29008, 2003.
21. Beatriz E. Rendon, Thierry Roger, Ivo Teneng, Ming Zhao, Yousef Al-Abed, Thierry Calandra, and Robert A. Mitchell. Regulation of Human Lung Adenocarcinoma Cell Migration and Invasion by Macrophage Migration Inhibitory Factor. J Biol Chem, 282: 29910-29918, 2007.
22. Lijun Xu and Xingming Deng. Suppression of Cancer Cell Migration and Invasion by Protein Phosphatase 2A through Dephosphorylation of μ- and m-Calpains. J Biol Chem, 281: 35567-35575, 2006.
23. S. Buckley, W. Shi, B. Driscoll, A. Ferrario, K. Anderson, and D. Warburton. BMP4 signaling induces senescence and modulates the oncogenic phenotype of A549 lung adenocarcinoma cells. Am J Physiol Lung Cell Mol Physiol, 286: L81-L86, 2004.
24. S. Buckley, B. driscoll, W. Shi, K. anderson, and D. Warburton. Migration and gelatinases in cultured fetal, adult, and hyperoxic alveolar epithelial cells. Am J Physiol Lung Cell Mol Physiol, 281: L427-L434, 2001.
25. Masahiro Sasaki, Takefumi Ito, Masayuki Kashima, Shin Fukui, Noriko Izumiyama, Akiko Watanabe, Masaaki Sano, Yosimasa Fujiwara and Mamoru Miura. Erythromycin and clarithromycin modulation of growth factor-induced expression of heparanase mRNA on human lung cancer cells in vitro. Mediators Inflamm, 10:259-267, 2001.
26. William P. Tew, David P. Kelsen, David H. Ilson. Targeted Therapies for Esophageal Cancer. The Oncologist, 590-601, 2005.
27. Noriko Ohno, Akiko Izawa, Masakazu Hattori, Ryoichiro Kageyama, Tetsuo Sudo. dlk Inhibits Stem Cell Factor-Induced Colony Formation of Murine Hematopoietic Progenitors: Hes-1-Independent Effect. Stem Cells, 19:71-79, 2001.
28. Haobin Chen and Max Costa. Effect of Soluble Nickel on Cellular Energy Metabolism in A549 Cells. Exp Biol Med, 231:1474-1480, 2006.
29. Xinliang Mao, Eric Seidlitz, Kakoli Ghosh, Yoshinori Murakami, and Hara P. Ghosh. The Cytoplasmic Domain Is Critical to the Tumor Suppressor Activity of TSLC1 in Non-Small Cell Lung Cancer. Cancer Res, 63:7979-7985, 2003.
30. A Preet, RK Ganju and JE Groopman. D9-Tetrahydrocannabinol inhibits epithelial growth factor-induced lung cancer cell migration in vitro as well as its growth and metastasis in vivo. Oncogene, 27:339–346, 2008.
31. Sajani S. Lakka, Ramesh Rajagopal, Mannari K. Rajan, Pamarthi M. Mohan, Yoshiaki Adachi, Dzung H. Dinh, et al. Adenovirus-mediated Antisense Urokinase-Type Plasminogen Activator Receptor Gene Transfer Reduces Tumor Cell Invasion and Metastasis in Non-Small Cell Lung Cancer Cell Lines. Clinical Cancer Research, 7: 1087–1093, 2001.
32. KAZUTAKA FUKUSHIMA, KATSUKIYO YAZAWA and TADASHI ARAI. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF SAFRAMYCINS, HETEROCYCLIC QUINONE ANTITUMOR ANTIBIOTICS. THE JOURNAL OF ANTIBIOTICS, 1602-1604, 1986.