跳到主要內容

臺灣博碩士論文加值系統

(3.235.185.78) 您好!臺灣時間:2021/07/27 17:29
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:李宗恒
研究生(外文):Tsung-Heng Lee
論文名稱:探討BO-1051,9-anilinoacridine N-mustard衍生物,誘導細胞自噬調控的細胞死亡與對口腔癌細胞產生毒性的機制
論文名稱(外文):BO-1051, a N-mustard derivatives of 9-anilinoacridine, induces autophagy-mediated cell death and its underlying mechanism of cytotoxicity in oral cancer cells
指導教授:羅正汎邱士華邱士華引用關係
指導教授(外文):Jeng-Fan LoShih-Hwa Chiou
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:口腔生物研究所
學門:醫藥衛生學門
學類:牙醫學類
論文種類:學術論文
畢業學年度:97
語文別:中文
論文頁數:77
中文關鍵詞:口腔癌
外文關鍵詞:BO-1051Autophagy
相關次數:
  • 被引用被引用:0
  • 點閱點閱:138
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
頭頸鱗狀上皮癌(Head and neck squamous cell carcinoma),包括口腔癌(oral squamous cell carcinoma),在常見人類癌症種類中為排名第六位。在東南亞國家,飲酒、抽菸與嚼食檳榔為腫瘤生成的主要高危險因子。從臨床治療的經驗得知,通常透過手術、放射與化學治療的口腔癌第三、四期病患,其五年存活率大約為50%,在近十年間並無明顯改善。BO-1051是一種新的9-anilinoacridine N-mustard衍生抗癌藥物,在人類腫瘤異種移植的裸鼠上能有效抑制腫瘤生長。在本研究中發現,當口腔癌細胞處理低劑量BO-1051的IC50值濃度時,產生的細胞毒性能抑制其生長並且誘導細胞週期停滯。而隨著checkpoint kinases的表現量上升顯示BO-1051能導致DNA損傷並且參與DNA的修復機制。另一方面,BO-1051會隨時間及劑量的上升而活化細胞自噬中的LC3 I-II蛋白。然而,處理24小時高劑量的BO-1051仍可誘導細胞凋亡中的caspase3及PARP表現量上升,表示未來應該利用細胞自噬的抑制劑3-MA或bafilomycin A1來測試死亡種類的機制順序。此外,BO-1051結合放射線照射雖然能延緩異種移植裸鼠的腫瘤生長,但無法完全產生放射線加成的效果。綜合以上結果顯示,BO-1051結合放射線治療在未來能當作治療口腔癌的重要療程。
Head and neck squamous cell carcinoma (HNSCC), including oral cancer, is the sixth most common types of human cancer. In South-Eastern Asia, alcohol, tobacco and areca nut abuse are main risk of carcinogenic factors. The five year survival rate of approximately 50% patients, particularly in stage III or IV, has not improved through surgery, radiotherapy or chemotherapy during decade. BO-1051 is a new 9-anilinoacridine N-mustard derivative and exhibits potential antitumor therapeutic efficacy in nude mice against human tumor xenogeafts. In this study, we found that treatment of BO-1051 in oral cancer cell lines induce significant cytotoxicity with low IC50 and cell cycle arrest. Data of up-regulated checkpoint kinases expression demonstrated that BO-1051 caused DNA damage and involved in mechanism of DNA repair. Our data also showed that the new compound could active autophagic cell death through expression of marked LC3II with time- and dose- dependent manner. However, BO-1051 might induce apoptosis over 24 hour by up-ragulated caspase 3 and PARP. It suggests that we have to examine the order of cell death-induced with autophagy inhibitors 3-MA or bafilomycin A1. On the other hand, we also found that BO-1051 could cause tumor growth delay, but not enhance the radiosensitivity of a SAS-derived xenograft. Together, these results suggest that BO-1051 may be a potential cancer agent combined with radiation for oral therapy.
論文電子檔著作權授權書......................................
論文審定同書................................................
致謝.......................................................1
中文摘要...................................................2
英文摘要...................................................3
目錄.......................................................4
圖目錄.....................................................8
第一章 緒言...............................................9
1、口腔癌.............................................10
2、BO-1051............................................14
3、Autophagy..........................................16
4、DNA 損傷反應.......................................21
第二章 研究目的..........................................22
第三章 研究材料及方法....................................26
1、細胞培養 (Cell culture)................................27
2、細胞數目計數...........................................27
3、細胞冷凍...............................................27
4、細胞解凍...............................................28
5、細胞活性 (Cell viability) .............................28
6、流式細胞儀偵測Cell cycle...............................28
7、流式細胞儀偵測AVO (Acidic vesicular organelles)........29
8、流式細胞儀偵測Apoptosis................................29
9、免疫螢光染色 (Immunofluorescence ).....................30
10、西方點墨法 (Western blotting).........................30
10.1、蛋白質萃取 (Protein extraction) ....................30
10.2、凝膠體電泳法 (Sodium dodecyl sulfate polyacrylamide
gel electrophoresis)..................31
10.3、蛋白質轉印法 (Electrophoretic transfer).............32
10.4、免疫染色法 (Immunoblotting).........................32
11、單細胞膠體電泳 (Single cell gel electrophoresis or
Comet assay)..........................33
12、腫瘤生長能力試驗 (Tumor growth delay assay)...........34
第四章 結果..............................................36
第五章 討論..............................................42
第六章 結論..............................................49
參考文獻..................................................51
圖........................................................59
附錄......................................................72
Anslow, W.P., Jr., Karnofsky, D.A., and et al. (1948). The intravenous, subcutaneous and cutaneous toxicity of bis (beta-chloroethyl) sulfide (mustard gas) and of various derivatives. J Pharmacol Exp Ther 93, 1-9.

Ausio, J., and Abbott, D.W. (2002). The many tales of a tail: carboxyl-terminal tail heterogeneity specializes histone H2A variants for defined chromatin function. Biochemistry 41, 5945-5949.

Baraldi, P.G., Romagnoli, R., Guadix, A.E., Pineda de las Infantas, M.J., Gallo, M.A., Espinosa, A., Martinez, A., Bingham, J.P., and Hartley, J.A. (2002). Design, synthesis, and biological activity of hybrid compounds between uramustine and DNA minor groove binder distamycin A. J Med Chem 45, 3630-3638.

Brendel, M., and Ruhland, A. (1984). Relationships between functionality and genetic toxicology of selected DNA-damaging agents. Mutat Res 133, 51-85.

Budach, W., Hehr, T., Budach, V., Belka, C., and Dietz, K. (2006). A meta-analysis of hyperfractionated and accelerated radiotherapy and combined chemotherapy and radiotherapy regimens in unresected locally advanced squamous cell carcinoma of the head and neck. BMC Cancer 6, 28.

Bursch, W., Ellinger, A., Kienzl, H., Torok, L., Pandey, S., Sikorska, M., Walker, R., and Hermann, R.S. (1996). Active cell death induced by the anti-estrogens tamoxifen and ICI 164 384 in human mammary carcinoma cells (MCF-7) in culture: the role of autophagy. Carcinogenesis 17, 1595-1607.

de Castro, G., Jr., Snitcovsky, I.M., Gebrim, E.M., Leitao, G.M., Nadalin, W., Ferraz, A.R., and Federico, M.H. (2007). High-dose cisplatin concurrent to conventionally delivered radiotherapy is associated with unacceptable toxicity in unresectable, non-metastatic stage IV head and neck squamous cell carcinoma. Eur Arch Otorhinolaryngol 264, 1475-1482.

Derk, C.T., Rasheed, M., Artlett, C.M., and Jimenez, S.A. (2006). A cohort study of cancer incidence in systemic sclerosis. J Rheumatol 33, 1113-1116.

Downs, J.A., Lowndes, N.F., and Jackson, S.P. (2000). A role for
Saccharomyces cerevisiae histone H2A in DNA repair. Nature 408,
1001-1004.

Fernandez-Capetillo, O., Lee, A., Nussenzweig, M., and Nussenzweig, A.
(2004). H2AX: the histone guardian of the genome. DNA Repair (Amst)
3, 959-967.

Fu, K.K., Pajak, T.F., Trotti, A., Jones, C.U., Spencer, S.A., Phillips, T.L., Garden, A.S., Ridge, J.A., Cooper, J.S., and Ang, K.K. (2000). A Radiation Therapy Oncology Group (RTOG) phase III randomized study to compare hyperfractionation and two variants of accelerated fractionation to standard fractionation radiotherapy for head and neck squamous cell carcinomas: first report of RTOG 9003. Int J Radiat Oncol Biol Phys 48, 7-16.

Gibson, M.K., Li, Y., Murphy, B., Hussain, M.H., DeConti, R.C., Ensley, J., and Forastiere, A.A. (2005). Randomized phase III evaluation of cisplatin plus fluorouracil versus cisplatin plus paclitaxel in advanced head and neck cancer (E1395): an intergroup trial of the Eastern Cooperative Oncology Group. J Clin Oncol 23, 3562-3567.

Gillison, M.L., Koch, W.M., Capone, R.B., Spafford, M., Westra, W.H., Wu, L., Zahurak, M.L., Daniel, R.W., Viglione, M., Symer, D.E., et al. (2000). Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst 92, 709-720.

Gourdie, T.A., Valu, K.K., Gravatt, G.L., Boritzki, T.J., Baguley, B.C., Wakelin, L.P., Wilson, W.R., Woodgate, P.D., and Denny, W.A. (1990). DNA-directed alkylating agents. 1. Structure-activity relationships for acridine-linked aniline mustards: consequences of varying the reactivity of the mustard. J Med Chem 33, 1177-1186.

Gunn, J.M., Clark, M.G., Knowles, S.E., Hopgood, M.F., and Ballard, F.J. (1977). Reduced rates of proteolysis in transformed cells. Nature 266, 58-60.

Haddad, R.I., and Shin, D.M. (2008). Recent advances in head and neck cancer. N Engl J Med 359, 1143-1154.

Hanahan, D., and Weinberg, R.A. (2000). The hallmarks of cancer. Cell
100, 57-70.

Horiot, J.C., Bontemps, P., van den Bogaert, W., Le Fur, R., van den Weijngaert, D., Bolla, M., Bernier, J., Lusinchi, A., Stuschke, M., Lopez-Torrecilla, J., et al. (1997). Accelerated fractionation (AF) compared to conventional fractionation (CF) improves loco-regional control in the radiotherapy of advanced head and neck cancers: results of the EORTC 22851 randomized trial. Radiother Oncol 44, 111-121.

Huang, X., Halicka, H.D., and Darzynkiewicz, Z. (2004). Detection of histone H2AX phosphorylation on Ser-139 as an indicator of DNA damage (DNA double-strand breaks). Curr Protoc Cytom Chapter 7, Unit 7 27.

Hurley, L.H. (2002). DNA and its associated processes as targets for cancer therapy. Nat Rev Cancer 2, 188-200.

Kanzawa, T., Germano, I.M., Komata, T., Ito, H., Kondo, Y., and Kondo, S. (2004). Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ 11, 448-457.

Kanzawa, T., Zhang, L., Xiao, L., Germano, I.M., Kondo, Y., and Kondo, S. (2005). Arsenic trioxide induces autophagic cell death in malignant glioma cells by upregulation of mitochondrial cell death protein BNIP3. Oncogene 24, 980-991.

Kapuriya, N., Kapuriya, K., Zhang, X., Chou, T.C., Kakadiya, R., Wu, Y.T., Tsai, T.H., Chen, Y.T., Lee, T.C., Shah, A., et al. (2008). Synthesis and biological activity of stable and potent antitumor agents, aniline nitrogen mustards linked to 9-anilinoacridines via a urea linkage. Bioorg Med Chem 16, 5413-5423.

Knaggs, S., Malkin, H., Osborn, H.M., Williams, N.A., and Yaqoob, P. (2005). New prodrugs derived from 6-aminodopamine and 4-aminophenol as candidates for melanocyte-directed enzyme prodrug therapy (MDEPT). Org Biomol Chem 3, 4002-4010.

Leach, T.J., Mazzeo, M., Chotkowski, H.L., Madigan, J.P., Wotring, M.G., and Glaser, R.L. (2000). Histone H2A.Z is widely but nonrandomly distributed in chromosomes of Drosophila melanogaster. J Biol Chem 275, 23267-23272.

Lee, H.K., Jones, R.T., Myers, R.A., and Marzella, L. (1992). Regulation of protein degradation in normal and transformed human bronchial epithelial cells in culture. Arch Biochem Biophys 296, 271-278.

Liang, X.H., Jackson, S., Seaman, M., Brown, K., Kempkes, B., Hibshoosh, H., and Levine, B. (1999). Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402, 672-676.

Lockshin, R.A., and Zakeri, Z. (2002). Caspase-independent cell deaths. Curr Opin Cell Biol 14, 727-733.

Mains, R.E., and May, V. (1988). The role of a low pH intracellular compartment in the processing, storage, and secretion of ACTH and endorphin. J Biol Chem 263, 7887-7894.

Majno, G., and Joris, I. (1995). Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146, 3-15.

Mannironi, C., Bonner, W.M., and Hatch, C.L. (1989). H2A.X. a histone isoprotein with a conserved C-terminal sequence, is encoded by a novel mRNA with both DNA replication type and polyA 3' processing signals. Nucleic Acids Res 17, 9113-9126.

McClean, S., Costelloe, C., Denny, W.A., Searcey, M., and Wakelin, L.P. (1999). Sequence selectivity, cross-linking efficiency and cytotoxicity of DNA-targeted 4-anilinoquinoline aniline mustards. Anticancer Drug Des 14, 187-204.

Meng, C.L., Tu, L.C., Chang, L.C., Hsu, S.F., Wu, C.H. (1985). Establishment and characterization of a human oral epidermoid carcinoma cell line. Molecular Biology of neoplasia(Academia sinica, Taipei 374-6.

Mignogna, M.D., Fedele, S., Lo Russo, L., Lo Muzio, L., and Bucci, E. (2004). Immune activation and chronic inflammation as the cause of malignancy in oral lichen planus: is there any evidence ? Oral Oncol 40, 120-130.

Mignogna, M.D., Fedele, S., Lo Russo, L., Mignogna, C., de Rosa, G., and Porter, S.R. (2007). Field cancerization in oral lichen planus. Eur J Surg Oncol 33, 383-389.

Molinolo, A.A., Amornphimoltham, P., Squarize, C.H., Castilho, R.M., Patel, V., and Gutkind, J.S. (2009). Dysregulated molecular networks in head and neck carcinogenesis. Oral Oncol 45, 324-334.

Munger, K., Werness, B.A., Dyson, N., Phelps, W.C., Harlow, E., and Howley, P.M. (1989). Complex formation of human papillomavirus E7 proteins with the retinoblastoma tumor suppressor gene product. EMBO J 8, 4099-4105.

Muramatsu, H., Kogawa, K., Tanaka, M., Okumura, K., Nishihori, Y., Koike, K., Kuga, T., and Niitsu, Y. (1995). Superoxide dismutase in SAS human tongue carcinoma cell line is a factor defining invasiveness and cell motility. Cancer Res 55, 6210-6214.

Ogier-Denis, E., and Codogno, P. (2003). Autophagy: a barrier or an adaptive response to cancer. Biochim Biophys Acta 1603, 113-128.

Otsuka, H., and Moskowitz, M. (1978). Differences in the rates of protein degradation in untrasformed and transformed cell lines. Exp Cell Res 112, 127-135.

Paglin, S., Hollister, T., Delohery, T., Hackett, N., McMahill, M., Sphicas, E., Domingo, D., and Yahalom, J. (2001). A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res 61, 439-444.

Parkin, D.M., Bray, F., Ferlay, J., and Pisani, P. (2005). Global cancer statistics, 2002. CA Cancer J Clin 55, 74-108.

Petrini, J.H., and Stracker, T.H. (2003). The cellular response to DNA double-strand breaks: defining the sensors and mediators. Trends Cell Biol 13, 458-462.

Pignon, J.P., Bourhis, J., Domenge, C., and Designe, L. (2000). Chemotherapy added to locoregional treatment for head and neck squamous-cell carcinoma: three meta-analyses of updated individual data. MACH-NC Collaborative Group. Meta-Analysis of Chemotherapy on Head and Neck Cancer. Lancet 355, 949-955.

Redon, C., Pilch, D., Rogakou, E., Sedelnikova, O., Newrock, K., and Bonner, W. (2002). Histone H2A variants H2AX and H2AZ. Curr Opin Genet Dev 12, 162-169.

Rogakou, E.P., Pilch, D.R., Orr, A.H., Ivanova, V.S., and Bonner, W.M. (1998). DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem 273, 5858-5868.

Scudiero, D.A., Shoemaker, R.H., Paull, K.D., Monks, A., Tierney, S., Nofziger, T.H., Currens, M.J., Seniff, D., and Boyd, M.R. (1988). Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res 48, 4827-4833.

Seiwert, T.Y., and Cohen, E.E. (2005). State-of-the-art management of locally advanced head and neck cancer. Br J Cancer 92, 1341-1348.

Shao, Y., Gao, Z., Marks, P.A., and Jiang, X. (2004). Apoptotic and autophagic cell death induced by histone deacetylase inhibitors. Proc Natl Acad Sci U S A 101, 18030-18035.

Skehan, P., Storeng, R., Scudiero, D., Monks, A., McMahon, J., Vistica, D., Warren, J.T., Bokesch, H., Kenney, S., and Boyd, M.R. (1990). New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82, 1107-1112.

Stucki, M., Clapperton, J.A., Mohammad, D., Yaffe, M.B., Smerdon, S.J., and Jackson, S.P. (2005). MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell 123, 1213-1226.

Sudbo, J., Kildal, W., Risberg, B., Koppang, H.S., Danielsen, H.E., and Reith, A. (2001). DNA content as a prognostic marker in patients with oral leukoplakia. N Engl J Med 344, 1270-1278.

Suzukake, K., Vistica, B.P., and Vistica, D.T. (1983). Dechlorination of L-phenylalanine mustard by sensitive and resistant tumor cells and its relationship to intracellular glutathione content. Biochem Pharmacol 32, 165-167.

Takeuchi, H., Kondo, Y., Fujiwara, K., Kanzawa, T., Aoki, H., Mills, G.B., and Kondo, S. (2005). Synergistic augmentation of rapamycin-induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors. Cancer Res 65, 3336-3346.

Traganos, F., and Darzynkiewicz, Z. (1994). Lysosomal proton pump activity: supravital cell staining with acridine orange differentiates leukocyte subpopulations. Methods Cell Biol 41, 185-194.

Werness, B.A., Levine, A.J., and Howley, P.M. (1990). Association of human papillomavirus types 16 and 18 E6 proteins with p53. Science 248, 76-79.

Yao, K.C., Komata, T., Kondo, Y., Kanzawa, T., Kondo, S., and Germano, I.M. (2003). Molecular response of human glioblastoma multiforme cells to ionizing radiation: cell cycle arrest, modulation of the expression of cyclin-dependent kinase inhibitors, and autophagy. J Neurosurg 98, 378-384.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top