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研究生:鄭宇辰
研究生(外文):Yu-Chen Cheng
論文名稱:前列腺癌抗腫瘤療法之研究
論文名稱(外文):The Study of Antitumor Therapy in Prostate Carcinoma
指導教授:陳正繹李奇峰李奇峰引用關係趙壯飛
指導教授(外文):Jang-Yi ChenChi-Feng LeeChung-Faye Chao
口試委員:趙壯飛李奇峰周成功
口試委員(外文):Chung-Faye ChaoChi-Feng LeeCheng-Kung Chou
口試日期:2011-05-27
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物及解剖學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:130
中文關鍵詞:前列腺癌抗腫瘤療法
外文關鍵詞:prostate cancersunitinibBCG
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在對抗癌症的免疫治療上,使用卡介苗(bacille Calmette-Gue’rin; BCG)免疫療法來治療表淺性膀胱移行上皮癌(TCC),在臨床上已被視為預防膀胱癌復發及治療殘留癌的第一線治療方法。 在動物模式中可發現,移植了腫瘤的小鼠在接受具免疫刺激能力的BCG治療後可誘發其體內的免疫反應,能產生抑制腫瘤生長及延長小鼠壽命的結果。 而在化學治療上,Sunitinib malate是一種抗腫瘤的藥物,他是一種針對多種酪胺酸激酶受體〔例如:血管生長因子受體(VEGFRs)、血小板衍生生長因子受體(PDGF-Rs)〕的抑制劑,其在動物實驗中證明能使腫瘤細胞凋亡並減少腫瘤周圍的血管新生,最終達成使腫瘤萎縮的效果。 藉於這一點,我們假設BCG疫苗株及Sunitinib malate的合併使用也許能成為一種新的治療前列腺癌的藥物及方式。 為了要證明這個假設,BCG疫苗株及Sunitinib malate在實驗中被用來測試其治療前列腺癌的效果;首先我們在測試及分析Sunitinib malate誘導腫瘤細胞凋亡及其對一些腫瘤生長因子(例如AKT、ERK)及下游凋亡因子(例如caspase-3、caspase-9)的影響,藉此證明了Sunitinib malate在前列腺癌細胞上同樣具有誘導腫瘤細胞凋亡及抑制腫瘤細胞生長的效果。 第二步則在移植腫瘤的動物模式中證明了BCG疫苗株及Sunitinib malate的合併使用能產生更為明顯的抑制腫瘤生長,造成腫瘤萎縮的效果。 在未來,這也許能成為一種新的有效治療前列腺癌的藥物及方式。
Mycobacterium bovis bacillus Calmette Guérin(BCG)-immunotherapy has a well-documented and successful clinical history in the treatment of superficial bladder transitional-cell carcinoma(TCC). Highly immunopotent BCG treatment elicited immune responses, inhibited tumor growth and prolonged survival in tumor bearing mice. Sunitinib malate an anti-cancer drug is a multi-targeted receptor tyrosine kinase inhibitor of VEGFRs and PDGFRs, leads to both reduce tumor vascularization and cause cancer cell to apoptosis, and ultimately made tumor shrinkage. Regarding this, we hypothesized that combined with BCG vaccine and sunitinib might be appreciate therapeutic strategy for prostate cancer. To evaluate this hypothesis, BCG subcomponents and sunitinib was used to test prostate cancer therapy. In the first step, we investigated the effectiveness of sunitinib on reducing both cell growth through modulating AKT/ERK signal transduction pathway and promoting cancer cell apoptosis in vitro and in vivo. In the following step, we had demonstrated that combined treatment of BCG vaccine and sunitinib had more effective on inhibition of tumor growth and ultimately cause tumor shrinkage in prostate carcinoma of a tumor-bearing animal model. Therefore, it might be a better strategy than treated BCG vaccine or sunitinib along for prostate cancer therapy.
本文目錄

本文目錄 I
圖表目錄 IV
附錄目錄 V
中文摘要 VI
英文摘要 VII

第一章 緒論 1
第一節 前列腺的結構與功能 1
第二節 前列腺癌的發生 2
第三節 前列腺癌的症狀 4
第四節 前列腺癌的流行病學 5
第五節 前列腺癌的診斷 6
第六節 前列腺癌的治療 8
第七節 前列腺癌治療方式的改進 11

第二章 實驗材料與方法 14
第一節 實驗材料 14
1. 實驗動物與細胞株 14
2. 實驗藥品與試劑 15
3. 實驗儀器 23
第二節 實驗方法 26
壹、 細胞培養、繼代培養與冷凍保存 26
一、 前列腺癌細胞 26
1. 大鼠前列腺癌細胞株AT3 26
2. 人類前列腺細胞癌雄性素依賴型細胞株LNCaP 27
3. 人類前列腺細胞癌雄性素非依賴型細胞株DU145 28
二、 細胞計數 29
三、 BCG菌的培養 29
貳、 細胞存活實驗 29
一、 以MTT分析Sunitinib對於癌細胞存活率之影響 29
二、 以細胞數量分析Sunitinib對於癌細胞的存活率影響 31
參、西方點墨法暨細胞免疫螢光染色 31
一、 西方點墨法 31
二、 細胞免疫螢光染色 35
肆、 Sunitinib對於腫瘤細胞生長調控因子的影響 36
伍、 細胞死亡分析 36
一、 細胞毒殺實驗 36
二、 細胞凋亡現象之觀察 37
三、 細胞凋亡途徑之分析 38
1. 以西方點墨法分析可能導致細胞死亡的相關路徑 38
2. 以Caspase-Glo Assay分析可能導致細胞死亡的路徑 38
陸、皮下注射DU145細胞模擬癌症術後復發之動物模式 39
一、 Sunitinib malate給藥方式 40
二、卡介苗(BCG):Tokyo strain 172菌株給藥方式 41

第三章 實驗結果 42
第一節 Sunitinib malate藥物對於前列腺癌細胞之存活率實驗 42
一、 以MTT分析Sunitinib對於癌細胞存活率之影響 42
二、 以細胞數量分析Sunitinib對於癌細胞的存活率影響 43
第二節 Sunitinib malate藥物對於腫瘤細胞生長調控因子的影響 45
一、 AKT之磷酸化現象 45
二、 ERK之磷酸化現象 47
第三節 Sunitinib malate藥物造成腫瘤細胞凋亡的影響及其途徑 49
一、 分析Sunitinib對於前列腺癌細胞的毒殺效果 49
二、 觀察Sunitinib是否會造成前列腺癌細胞之凋亡 51
三、 以西方點墨法觀察Sunitinib對於前列腺癌細胞凋亡之
影響及途徑 52
1. p38 MAPK之磷酸化現象 52
2. JNK之磷酸化現象 54
3. Caspase-3活化之現象 56
4. Caspase-9活化之現象 58
四、 以Caspase-Glo Assay分析Sunitinib對於前列腺癌細胞
凋亡之影響及途徑 60
1. Caspase-3活化之現象 60
2. Caspase-9活化之現象 62
第四節 皮下注射DU145細胞模擬前列腺癌術後復發之動物模式 64
一、 腫瘤體積大小之變化 64
二、 小鼠體重大小之變化 65

第四章 討論 66
第一節 Sunitinib malate藥物對前列癌細胞之影響 67
一、 Sunitinib對前列腺癌細胞存活率之影響 67
二、 Sunitinib對前列腺癌細胞生長調控因子之影響 68
三、 Sunitinib造成前列腺癌細胞死亡之影響 68
四、 Sunitinib造成前列腺癌細胞凋亡之影響 69
五、 Sunitinib造成前列腺癌細胞凋亡之途徑 70
第二節 動物模式中標靶藥物Sunitinib malate合併BCG卡介苗疫
苗免疫治療之療效 71

第五章 圖表結果 74

第六章 參考文獻 104

附錄

1.Abouantoun TJ, Castellino RC, MacDonald TJ:Sunitinib induces PTEN expression and inhibits PDGFR signaling and migration of medulloblastoma cells. J Neurooncol. 101(2):215-26, 2010.
2.American Cancer Society webpage. Detailed Guide:prostate cancer. 2010.
3.Aragon-Ching JB, Dahut WL:About tyrosine kinase inhibitors (TKIs) in prostate cancer: where do we go from here? Ann Oncol. 21(1):183-4, 2010.
4.Aumüller, G. Prostate Gland and Seminal Vesicles. Springer-Verlag Berlin-Heidelberg. 1979.
5.Bachleda P, Dvorák Z:Pharmacological inhibitors of JNK and ERK kinases SP600125 and U0126 are not appropriate tools for studies of drug metabolism because they activate aryl hydrocarbon receptor. Gen Physiol Biophys. 27(2):143-5, 2008.
6.Burstein HJ, Elias AD, Rugo HS, Cobleigh MA, Wolff AC, Eisenberg PD, Lehman M, Adams BJ, Bello CL, DePrimo SE, Baum CM, Miller KD:Phase II study of sunitinib malate, an oral multitargeted tyrosine kinase inhibitor, in patients with metastatic breast cancer previously treated with an anthracycline and a taxane. J Clin Oncol. 26:1810-1816, 2008.
7.Castillo-Avila W, Piulats JM, Garcia Del Muro X, Vidal A, Condom E, Casanovas O, Mora J, Germa JR, Capella G, Villanueva A, Vinals F:Sunitinib inhibits tumor growth and synergizes with cisplatin in orthotopic models of cisplatin-sensitive and cisplatin-resistant human testicular germ cell tumors. Clin Cancer Res. 15:3384-3395, 2009.
8.Christophe LT, Eric R, Sandrine F:Sunitinib: a novel tyrosine kinase inhibitor. A brief review of its therapeutic potential in the treatment of renal carcinoma and gastrointestinal stromal tumors (GIST). TCRM. 3(2):341–348, 2007.
9.Chu TF, Rupnick MA, Kerkela R, Dallabrida SM, Zurakowski D, Nguyen L, Woulfe K, Pravda E, Cassiola F, Desai J, George S, Morgan JA, Harris DM, Ismail NS, Chen JH, Schoen FJ, Van den Abbeele AD, Demetri GD, Force T, Chen MH:Cardiotoxicity associated with tyrosine kinase inhibitor sunitinib. Lancet. 370(9604):2011-9, 2007.
10.Costa-Pereira AP, McKenna SL, Cotter TG:Activation of SAPK/JNK by camptothecin sensitizes androgen-independent prostate cancer cells to Fas-induced apoptosis. Br J Cancer. 82(11):1827-34, 2000.
11.Cumashi A, Tinari N, Rossi C, Lattanzio R, Natoli C, Piantelli M, Iacobelli S:Sunitinib malate:SU-11248 alone or in combination with low-dose docetaxel inhibits the growth of DU-145 prostate cancer xenografts. Cancer Lett. 270:229-233, 2008.
12.Czabanka M, Vinci M, Heppner F, Ullrich A, Vajkoczy P:Effects of sunitinib on tumor hemodynamics and delivery of chemotherapy. Int J Cancer. 124:1293-1300, 2009.
13.Deprimo SE, Bello CL, Smeraglia J, Baum CM, Spinella D, Rini BI, Michaelson MD, Motzer RJ:Circulating protein biomarkers of pharmacodynamic activity of sunitinib in patients with metastatic renal cell carcinoma: modulation of VEGF and VEGF-related proteins. J Transl Med. 5:32, 2007.
14.De S, Chen J, Narizhneva NV, Heston W, Brainard J, Sage EH, Byzova TV:Molecular pathway for cancer metastasis to bone. J Biol Chem. 278(40):39044-50, 2003.
15.Ebos JM, Lee CR, Christensen JG, Mutsaers AJ, Kerbel RS:Multiple circulating proangiogenic factors induced by sunitinib malate are tumor-independent and correlate with antitumor efficacy. Proc Natl Acad Sci U S A. 104:17069-17074, 2007.
16.Faivre S, Demetri G, Sargent W, Raymond E:Molecular basis for sunitinib efficacy and future clinical development. Nat Rev Drug Discov. 6:734-745, 2007.
17.Fizazi K, Sternberg CN, Fitzpatrick JM, Watson RW, Tabesh M:Role of targeted therapy in the treatment of advanced prostate cancer. BJU Int. 105(6):748-67, 2010.
18.Force T, Krause DS, Van Etten RA:Molecular mechanisms of cardiotoxicity of tyrosine kinase inhibition. Nat Rev Cancer. 7(5):332-44, 2007.
19.Giannopoulou E, Dimitropoulos K, Argyriou AA, Koutras AK, Dimitrakopoulos F, Kalofonos HP:An in vitro study, evaluating the effect of sunitinib and/or lapatinib on two glioma cell lines. Invest New Drugs. 2009.
20.Hankey BF, Feuer EJ, Clegg LX, Hayes RB, Legler JM, Prorok PC, Ries LA, Merrill RM, Kaplan RS. Cancer surveillance series:interpreting trends in prostate cancer--part I:Evidence of the effects of screening in recent prostate cancer incidence, mortality, and survival rates. J Natl Cancer Inst, 91(12):1017-24, 1999.
21.Hilchie AL, Furlong SJ, Sutton K, Richardson A, Robichaud MR, Giacomantonio CA, Ridgway ND, Hoskin DW:Curcumin-induced apoptosis in PC3 prostate carcinoma cells is caspase-independent and involves cellular ceramide accumulation and damage to mitochondria. Nutr Cancer. 62(3):379-89, 2010.
22.Hill KM, Kalifa S, Das JR, Bhatti T, Gay M, Williams D, Taliferro-Smith L, De Marzo AM:The role of PI 3-kinase p110beta in AKT signally, cell survival, and proliferation in human prostate cancer cells. Prostate. 70(7):755-64, 2010.
23.Hoffman RM;Gilliland FD;Eley JW;Harlan LC;Stephenson RA;Stanford JL;Albertson PC;Hamilton AS;Hunt WC;Potosky AL. Racial and ethnic differences in advanced-stage prostate cancer:the Prostate Cancer Outcomes Study. J Natl Cancer Inst. 93(5):388-95, 2001.
24.Homsi J, Daud AI:Spectrum of activity and mechanism of action of VEGF/PDGF inhibitors. Cancer Control. 14:285-294, 2007.
25.Hong Kong Urological Association webpage. Detailed Guide:prostate cancer. 2010.
26.Hu H, Jiang C, Li G, Lü J:PKB/AKT and ERK regulation of caspase-mediated apoptosis by methylseleninic acid in LNCaP prostate cancer cells. Carcinogenesis. 26(8):1374-81, 2005.
27.Huang D, Ding Y, Luo WM, Bender S, Qian CN, Kort E, Zhang ZF, VandenBeldt K, Duesbery NS, Resau JH, Teh BT:Inhibition of MAPK kinase signaling pathways suppressed renal cell carcinoma growth and angiogenesis in vivo. Cancer Res. 68(1):81-8, 2008.
28.Huang D, Ding Y, Li Y, Luo WM, Zhang ZF, Snider J, Vandenbeldt K, Qian CN, Teh BT:Sunitinib acts primarily on tumor endothelium rather than tumor cells to inhibit the growth of renal cell carcinoma. Cancer Res. 70(3):1053-62, 2010.
29.Indelicato M, Pucci B, Schito L, Reali V, Aventaggiato M, Mazzarino MC, Stivala F, Fini M, Russo MA, Tafani M:Role of hypoxia and autophagy in MDA-MB-231 invasiveness. J Cell Physiol. 223:359-368, 2010.
30.Jeong YM, Li H, Kim SY, Park WJ, Yun HY, Baek KJ, Kwon NS, Jeong JH, Myung SC, Kim DS:Photo-activated 5-hydroxyindole-3-acetic acid induces apoptosis of prostate and bladder cancer cells. J Photochem Photobiol B. 103(1):50-6, 2011.
31.Kamba T, McDonald DM:Mechanisms of adverse effects of anti-VEGF therapy for cancer. Br J Cancer. 96:1788-1795, 2007.
32.Kinkade CW, Castillo-Martin M, Puzio-Kuter A, Yan J, Foster TH, Gao H, Sun Y, Ouyang X, Gerald WL, Cordon-Cardo C, Abate-Shen C:Targeting AKT/mTOR and ERK MAPK signaling inhibits hormone-refractory prostate cancer in a preclinical mouse model. J Clin Invest. 118(9):3051-64, 2008.
33.Kumar R, Srinivasan S, Pahari P, Rohr J, Damodaran C:Activating stress-activated protein kinase-mediated cell death and inhibiting epidermal growth factor receptor signaling: a promising therapeutic strategy for prostate cancer. Mol Cancer Ther. 9(9):2488-96, 2010.
34.Lee CF, Chang SY, Hsieh DS, Yu DS:Treatment of bladder carcinomas using recombinant BCG DNA Vaccines and electroporative gene immunotherapy. Cancer Gene Therapy, 11:194-207, 2004.
35.Lee CF and Yu DS:BCG DNA Cancer Vaccine, US PATENT No. US7, 605, 139 B2, 2009.
36.Lee EY, Xia Y, Kim WS, Kim MH, Kim TH, Kim KJ, Park BS, Sung JH:Hypoxia-enhanced wound-healing function of adipose-derived stem cells:increase in stem cell proliferation and up-regulation of VEGF and bFGF. Wound Repair Regen. 17:540-547, 2009.
37.Li ZD, Bork JP, Krueger B, Patsenker E, Schulze-Krebs A, Hahn EG, Schuppan D:VEGF induces proliferation, migration, and TGF-beta1 expression in mouse glomerular endothelial cells via mitogen-activated protein kinase and phosphatidylinositol 3-kinase. Biochem Biophys Res Commun. 334:1049-1060, 2005.
38.Meshki J, Caino MC, von Burstin VA, Griner E, Kazanietz MG:Regulation of prostate cancer cell survival by protein kinase Cepsilon involves bad phosphorylation and modulation of the TNFalpha/JNK pathway. J Biol Chem. 285(34):26033-40, 2010.
39.Miller, DC;Hafez, KS, Stewart, A, et al. Prostate carcinoma presentation, diagnosis, and staging:an update form the National Cancer Data Base. Cancer, 98:1169, 2003.
40.Moore, K., Dalley, A. Clinically Oriented Anatomy. Lippincott Williams & Wilkins, Baltimore, Maryland. 1999.
41.Moulder JE, Rockwell S:Tumor hypoxia:its impact on cancer therapy. Cancer Metastasis Rev. 5:313-341, 1987.
42.Park IJ, Kim MJ, Park OJ, Park MG, Choe W, Kang I, Kim SS, Ha J:Cryptotanshinone sensitizes DU145 prostate cancer cells to Fas(APO1/CD95)-mediated apoptosis through Bcl-2 and MAPK regulation. Cancer Lett. 298(1):88-98, 2010.
43.Pedram A, Razandi M, Hu RM, Levin ER:Vasoactive peptides modulate vascular endothelial cell growth factor production and endothelial cell proliferation and invasion. J Biol Chem. 272:17097-17103, 1997.
44.Ping SY, Wu CL, Yu DS:Sunitinib can enhance BCG mediated cytotoxicity to transitional cell carcinoma through apoptosis pathway. Urol Oncol. 2010.
45.Rasola A, Sciacovelli M, Chiara F, Pantic B, Brusilow WS, Bernardi P:Activation of mitochondrial ERK protects cancer cells from death through inhibition of the permeability transition. Proc Natl Acad Sci U S A. 107(2):726-31, 2010.
46.Sarmishtha D, Olga R, Noel PM, Timothy OT, Jun Q, and Tatiana VB:VEGF–integrin interplay controls tumor growth and vascularization. PNAS. 102(21):7589–7594, 2005.
47.Schulze-Bergkamen H, Weinmann A, Moehler M, Siebler J, Galle PR:Novel ways to sensitise gastrointestinal cancer to apoptosis. Gut. 58(7):1010-24, 2009.
48.Smith ND, Rubenstein JN, Eggener SE, Kozlowski JM:The p53 tumor suppressor gene and nuclear protein:basic science review and relevance in the management of bladder cancer. J Urol. 169:1219-1228, 2003.
49.Sonpavde G, Jian W, Liu H, Wu MF, Shen SS, Lerner SP:2009 Sunitinib malate is active against human urothelial carcinoma and enhances the activity of cisplatin in a preclinical model. In: Urol Oncol, vol. 27, pp 391-399.
50.Sophie B, Paulette H, James GC, Edwige L, Pascal G, Eric R, Jean SG:Antiangiogenic and anti-invasive effects of sunitinib on experimental human glioblastoma. Neuro Oncol. 9(4):412-423, 2007.
51.Steive, H. Männliche Genitalorgane. In:Handbuch der mikroskopischen Anatomie des Menschen. Vol. VII Part 2, pp. 1-399. Berlin:Springer 1930.
52.Streeter EH, Harris AL:Angiogenesis in bladder cancer--prognostic marker and target for future therapy. Surg Oncol. 11:85-100, 2002.
53.Sun JC, Beilke JN, Lanier LL:Adaptive immune features of natural killer cells. Nature. 457(7229):557-61, 2009.
54.Sun JC, Lopez-Verges S, Kim CC, DeRisi JL, Lanier LL:NK cells and immune "memory". J Immunol. 186(4):1891-7, 2011.
55.Sykiotis GP, Bohmann D:Stress-Activated Protein Kinases. Heidelberg. Springer Berlin. 225-241, 2008.
56.Tanaka Y, Gavrielides MV, Mitsuuchi Y, Fujii T, Kazanietz MG:Protein kinase C promotes apoptosis in LNCaP prostate cancer cells through activation of p38 MAPK and inhibition of the Akt survival pathway. J Biol Chem. 278(36):33753-62, 2003.
57.Ugolini S, Vivier E:Immunology: Natural killer cells remember. Nature. 457(7229):544-5, 2009.
58.Van der Cruijsen-Koeter IW, Vis AN, Roobol MJ, Wildhagen MF, de Koning HJ, van der Kwast TH, Schroder FH. Comparison of screen detected and clinically diagnosed prostate cancer in the European randomized study of screening for prostate cancer, section rotterdam. Urol, 174(1):121-5 , 2005.
59.Vogl UM, Berger W, Micksche M, Pirker C, Lamm W, Pichelmeyer O, Zielinski CC, Schmidinger M:Synergistic effect of Sorafenib and Sunitinib with Enzastaurin, a selective protein kinase C inhibitor in renal cell carcinoma cell lines. Cancer Lett. 277:218-226, 2009.
60.Wagner EF, Nebreda AR:Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 9(8):537-49, 2009.
61.Weston CR, Davis RJ:The JNK signal transduction pathway. Curr Opin Cell Biol. 19 (2): 142-149, 2007.
62.Wykoff CC, Beasley NJ, Watson PH, Turner KJ, Pastorek J, Sibtain A, Wilson GD, Turley H, Talks KL, Maxwell PH, Pugh CW, Ratcliffe PJ, Harris AL:Hypoxia-inducible expression of tumor-associated carbonic anhydrases. Cancer Res. 60:7075-7083, 2000.
63.Xin H, Zhang C, Herrmann A, Du Y, Figlin R, Yu H:Sunitinib inhibition of Stat3 induces renal cell carcinoma tumor cell apoptosis and reduces immunosuppressive cells. Cancer Res. 69:2506-2513, 2009.
64.Yap TA, Garrett MD, Walton MI, Raynaud F, de Bono JS, Workman P:Targeting the PI3K-AKT-mTOR pathway: progress, pitfalls, and promises. Curr Opin Pharmacol. 8(4):393-412, 2008.
65.Yu DS, Tao MH, Chang SY, Lee CF:Antitumor effects of Bacillus Calmette-Guerin recombinant protein Mpt-64 on xenografted murine bladder cancer. The FEBS Journal, 275 Suppl 1:411, 2008.
66.Zeng Y, Opeskin K, Goad J, Williams ED:Tumor-induced activation of lymphatic endothelial cells via vascular endothelial growth factor receptor-2 is critical for prostate cancer lymphatic metastasis. Cancer Res. 66(19):9566-75,2006.

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