研究生: |
林楷涵 Kai-Han Lin |
---|---|
論文名稱: |
三環藥物Teroxirone對非小細胞肺癌細胞增生機制的探討 The Mechanistic Study of Growth Inhibition by Teroxirone in Human Non-Small Cell Lung Cancer Cells |
指導教授: |
方剛
Fang, Kang |
學位類別: |
碩士 Master |
系所名稱: |
生命科學系 Department of Life Science |
論文出版年: | 2012 |
畢業學年度: | 100 |
語文別: | 中文 |
論文頁數: | 78 |
中文關鍵詞: | Teroxirone 、DNA損傷 、細胞凋亡 |
英文關鍵詞: | Teroxirone, DNA damage, apoptosis |
論文種類: | 學術論文 |
相關次數: | 點閱:214 下載:10 |
分享至: |
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報 |
肺癌細胞依其組織型態可分為兩類:小細胞肺癌 (small cell lung cancer, SCLC) 以及非小細胞肺癌 (non-small cell lung cancer, NSCLC),而其中大多數屬於非小細胞肺癌。目前在臨床治療上經常使用的抗癌藥物 (例如: camptothecin, doxorubicin, etoposide, cisplatin) 容易產生抗藥性,導致治療的困難,因此本文開發目前臨床上還未徹底了解,但具有抗癌效果的化合物。
研究目標是發展小分子且低劑量的三環氧化物teroxirone,了解teroxirone如何引發細胞凋亡。實驗以MTT及細胞群落分析 (colony formation assay) 確認teroxirone確實能降低非小細胞肺癌的存活率。接著要確認非小細胞肺癌細胞的存活率下降的確是由於細胞凋亡所造成。再利用單細胞凝膠電泳 (single cell gel electrophoresis (SCGE) / comet assay),偵測teroxirone是否造成DNA 損傷。再利用propidium iodide (PI) 染色觀察teroxirone對細胞週期的影響,再利用二維流式細胞儀及DNA片段化實驗觀察teroxirone所誘導的細胞凋亡。
實驗結果顯示,teroxirone都會對NSCLC細胞造成不同程度DNA損傷,且細胞凋亡與p53的基因型有關。
Lung cancer according to biological characteristics can be classified two types: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), most of which belong to non-small cell lung cancer. Clinical treatment used anticancer drugs (ex: camptothecin, doxorubicin, etoposide, cisplatin) frequently is easy to become resistant, leading to difficulties of treatment. The purpose of the thesis is to study the effectiveness of potential anticancer drugs.
The goal of this study is to understand how low doses of teroxirone induce NSCLC cell apoptosis. Teroxirone can reduce the proliferation rate of non-small cell lung cancer on indicated by MTT assay and colony forming assay. To confirm the reduced NSCLC cell growth rate is indeed due to cell death. To be have used the single cell gel electrophoresis (SCGE) / comet assay to detect DNA damage. To be have used propidium iodide (PI) staining to observe that impact of teroxirone on cell cycle and the induced apoptosis by flow cytometry. Double staining with PI and Annexin V – FITC of cell death for apoptosis. At the same time, DNA fragmentation also confirmed teroxirone indeed induced cells apoptosis.
The experimental results showed that teroxirone caused different degrees of DNA damage in NSCLC cells, but is caused apoptosis only occurred in p53 wild-type cells.
Ames, M.M., Kovach, J.S. and Rubin, J. (1984) Pharmacological characterization of teroxirone, a triepoxide antitumor agent, in rats, rabbits, and humans, Cancer Research, 44, 4151-4156.
Ashkenazi, A. and Dixit, V.M. (1999) Apoptosis control by death and decoy receptors, Current Opinion in Cell Biology, 11, 255-260.
Atassi, G., et al. (1980) Antitumoral effect in mice of a new triepoxyde derivative: 1, 3, 5-triglycidyl-s-triazinetrione (NSC 296934), European Journal of Cancer, 16, 1561-1567.
Baldi, A., et al. (2011) Tumor suppressors and cell-cycle proteins in lung cancer, Pathology Research International, Article ID 605042.
Bonior, J., et al. (2012) Long-lasting effect of infant rats endotoxemia on heat shock protein 60 in the pancreatic acinar cells: involvement of toll-like receptor 4, International Journal of Inflammation, Article ID 354904.
Chatelut, E. (2011) Pharmacology of platinum compounds: differences between the three molecules and factors of interpatient variability, Bulletin du Cancer, 98, 1253-1261.
Cryns, V. and Yuan, J. (1998) Proteases to die for, Genes & development, 12, 1551-1570.
Dibb, M. and Ang, Y.S. (2011) Targeting the cell cycle in esophageal adenocarcinoma: an adjunct to anticancer treatment, World Journal of Gastroenterology, 17, 2063-2069.
Ding, D., et al. (2011) Cisplatin ototoxicity in rat cochlear organotypic cultures, Hearing Research, 282, 196-203.
Duarte, M.L., et al. (2009) Role of p53 in the induction of cyclooxygenase-2 by cisplatin or paclitaxel in non-small cell lung cancer cell lines, Cancer Letters, 279, 57-64.
Falchook, G.S., et al. (2011) Age-stratified phase I trial of a combination of bortezomib, gemcitabine, and liposomal doxorubicin in patients with advanced malignancies, Cancer Chemotherapy Pharmacology, 69, 1117-1126 .
Faleiro, L., et al. (1997) Multiple species of CPP32 and Mch2 are the major active caspases present in apoptotic cells, The EMBO Journal, 16, 2271-2281.
Feng, B., Wang, R. and Chen, L.B. (2011) MiR-100 resensitizes docetaxel-resistant human lung adenocarcinoma cells (SPC-A1) to docetaxel by targeting Plk1, Cancer Letters, 317, 184-191.
Fidler, J.M., et al. (2003) PG490-88, a derivative of triptolide, causes tumor regression and sensitizes tumors to chemotherapy, Molecular Cancer Therapeutic, 2, 855-862.
Galano, G., et al. (2011) Efficacy and tolerability of vinorelbine in the cancer therapy, Current Drug Safety, 6, 185-193.
Giannakakou, P., et al. (2001) Low concentrations of paclitaxel induce cell type-dependent p53, p21 and G1/G2 arrest instead of mitotic arrest: molecular determinants of paclitaxel-induced cytotoxicity, Oncogene, 20, 3806-3813.
Giono, L.E. and Manfredi, J.J. (2006) The p53 tumor suppressor participates in multiple cell cycle checkpoints, Journal of Cellular Physiology, 209, 13-20.
Guo, M., et al. (2004) Hypermethylation of the GATA genes in lung cancer, Clinical Cancer Research, 10, 7917-7924.
Halazonetis, T.D., Gorgoulis, V.G. and Bartek, J. (2008) An oncogene-induced DNA damage model for cancer development, Science, 319, 1352-1355.
Hanahan, D. and Weinberg, R.A. (2000) The hallmarks of cancer, Cell, 100, 57-70.
Harper, J.W. and Elledge, S.J. (2007) The DNA damage response: ten years after, Molecular Cell, 28, 739-745.
Hattangadi, D.K., et al. (2004) Influence of p53 and caspase 3 activity on cell death and senescence in response to methotrexate in the breast tumor cell, Biochemical Pharmacology, 68, 1699-1708.
Hengartner, M.O. (2000) The biochemistry of apoptosis, Nature, 407, 770-776.
Hsieh, J.L., et al. (2012) Acquisition of an enhanced aggressive phenotype in human lung cancer cells selected by suboptimal doses of cisplatin following cell deattachment and reattachment, Cancer Letters, 321, 36-44.
Huang, D.C. and Strasser, A. (2000) BH3-Only proteins-essential initiators of apoptotic cell death, Cell, 103, 839-842.
Huang, P., et al. (1991) Action of 2',2'-difluorodeoxycytidine on DNA synthesis, Cancer Research, 51, 6110-6117.
Huisman, C., et al. (2002) Paclitaxel triggers cell death primarily via caspase-independent routes in the non-small cell lung cancer cell line NCI-H460, Clinical Cancer Research, 8, 596-606.
Inoue, A., et al. (2006) Prospective phase II study of gefitinib for chemotherapy-naive patients with advanced non-small-cell lung cancer with epidermal growth factor receptor gene mutations, Joural Clinical Oncology, 24, 3340-3346.
Jackson, S.P. and Bartek, J. (2009) The DNA-damage response in human biology and disease, Nature, 461, 1071-1078.
Jiang, X.H., et al. (2001) Functional p53 is required for triptolide-induced apoptosis and AP-1 and nuclear factor-kappaB activation in gastric cancer cells, Oncogene, 20, 8009-8018.
Jinturkar, K.A., et al. (2012) Liposomal formulations of Etoposide and Docetaxel for p53 mediated enhanced cytotoxicity in lung cancer cell lines, Biomaterials, 33, 2492-2507.
Johnson, J.R., et al. (2005) Approval summary for erlotinib for treatment of patients with locally advanced or metastatic non-small cell lung cancer after failure of at least one prior chemotherapy regimen, Clinical Cancer Research, 11, 6414-6421.
Kelly, G.L. and Strasser, A. (2011) The essential role of evasion from cell death in cancer, Advances in Cancer Research, 111, 39-96.
Kiviharju, T.M., et al. (2002) Antiproliferative and proapoptotic activities of triptolide (PG490), a natural product entering clinical trials, on primary cultures of human prostatic epithelial cells, Clinical Cancer Resaerch, 8, 2666-2674.
Kobayashi, T., et al. (2012) A phase II trial of erlotinib in patients with EGFR wild-type advanced non-small-cell lung cancer, Cancer Chemotherapy Pharmacology, 69, 1241-1246.
Lee, Y.R., et al. (2012) Reversine suppresses oral squamous cell carcinoma via cell cycle arrest and concomitantly apoptosis and autophagy, Journal of Biomedical Science, 19, 9-20.
Li, R., et al. (2005) Apoptosis of non-small-cell lung cancer cell lines after paclitaxel treatment involves the BH3-only proapoptotic protein Bim, Cell Death and Differentiation, 12, 292-303.
Lyseng-Williamson, K.A. and Fenton, C. (2005) Docetaxel: a review of its use in metastatic breast cancer, Drugs, 65, 2513-2531.
Maemondo, M., et al. (2010) Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR, The New England Journal of Medicine, 362, 2380-2388.
Malumbres, M. and Barbacid, M. (2009) Cell cycle, CDKs and cancer: a changing paradigm, Nature Reviews Cancer, 9, 153-166.
Martins, L.M., et al. (1997) Activation of multiple interleukin-1beta converting enzyme homologues in cytosol and nuclei of HL-60 cells during etoposide-induced apoptosis, The Journal of Biological Chemistry, 272, 7421-7430.
Mashima, T., et al. (1995) Aspartate-based inhibitor of interleukin-1 beta-converting enzyme prevents antitumor agent-induced apoptosis in human myeloid leukemia U937 cells, Biochemical and Biophysical Research Communications, 209, 907-915.
Mayo, L.D., et al. (2005) Phosphorylation of human p53 at serine 46 determines promoter selection and whether apoptosis is attenuated or amplified, The Journal of Biological Chemistry, 280, 25953-25959.
Mirshahidi, H.R. and Hsueh, C.T. (2010) Updates in non-small cell lung cancer - insights from the 2009 45(th) annual meeting of the American Society of Clinical Oncology, Journal Hematoology &Oncology, 3, 18-29.
Mok, T.S., et al. (2009) Gefitinib or carboplatin-paclitaxel in pulmonary adenocarcinoma, The New England Journal of Medicine, 361, 947-957.
Moretti, M., et al. (2011) A study protocol for the evaluation of occupational mutagenic/carcinogenic risks in subjects exposed to antineoplastic drugs: a multicentric project, BMC Public Health, 11, 195-204.
Muchmore, S.W., et al. (1996) X-ray and NMR structure of human Bcl-xL, an inhibitor of programmed cell death, Nature, 381, 335-341.
Neidhart, J.A., et al. (1984) Phase I trial of teroxirone, Cancer Treatment Reports, 68, 1115-1119.
Nicholson, D.W., et al. (1995) Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis, Nature, 376, 37-43.
Nodera, H., et al. (2011) Neuroprotective effects of Kv7 channel agonist, retigabine, for cisplatin-induced peripheral neuropathy, Neuroscience Letters, 505, 223-227.
Olewinska, E., et al. (2010) Level of DNA damage in lead-exposed workers, Annals of Agricultural and Environmental Medicine, 17, 231-236.
Osaki, S., et al. (2000) Alteration of drug chemosensitivity caused by the adenovirus-mediated transfer of the wild-type p53 gene in human lung cancer cells, Cancer Gene Therapy, 7, 300-307.
Patel, T., Gores, G.J. and Kaufmann, S.H. (1996) The role of proteases during apoptosis, The FASEB Journal, 10, 587-597.
Pierceall, W.E., et al. (2012) Cisplatin benefit is predicted by immunohistochemical analysis of DNA repair proteins in squamous cell carcinoma but not adenocarcinoma: theranostic modeling by NSCLC constituent histological subclasses, Annals of Oncology, 23, 2245-2252.
Plunkett, W., Huang, P. and Gandhi, V. (1995) Preclinical characteristics of gemcitabine, Anti-Cancer Drugs, 6, 7-13.
Plunkett, W., et al. (1995) Gemcitabine: metabolism, mechanisms of action, and self-potentiation, Seminars in Oncology, 22, 3-10.
Polverino, A.J. and Patterson, S.D. (1997) Selective activation of caspases during apoptotic induction in HL-60 cells. Effects of a tetrapeptide inhibitor, The Journal of Biological Chemistry, 272, 7013-7021.
Porter, A.G. and Janicke, R.U. (1999) Emerging roles of caspase-3 in apoptosis, Cell Death and Differentiation, 6, 99-104.
Sahu, B.D., et al. (2011) Carnosic acid attenuates renal injury in an experimental model of rat cisplatin-induced nephrotoxicity, Food and Chemical Toxicology, 49, 3090-3097.
Salvesen, G.S. and Dixit, V.M. (1997) Caspases: intracellular signaling by proteolysis, Cell, 91, 443-446.
Satyanarayana, A. and Kaldis, P. (2009) Mammalian cell-cycle regulation: several Cdks, numerous cyclins and diverse compensatory mechanisms, Oncogene, 28, 2925-2939.
Segawa, Y., et al. (2011) Small cell carcinoma of the tonsil treated with irinotecan and Cisplatin: a case report and literature review, Case Reports in Oncology, 4, 587-591.
Selvendiran, K., et al. (2010) Anticancer efficacy of a difluorodiarylidenyl piperidone (HO-3867) in human ovarian cancer cells and tumor xenografts, Molcular Cancer Thereputics, 9, 1169-1179.
Sengupta, S., et al. (2000) Etoposide encapsulated in positively charged liposomes: pharmacokinetic studies in mice and formulation stability studies, Pharmacological Research, 42, 459-464.
Shamon, L.A., et al. (1997) Evaluation of the mutagenic, cytotoxic, and antitumor potential of triptolide, a highly oxygenated diterpene isolated from Tripterygium wilfordii, Cancer Letters, 112, 113-117.
Shepherd, F.A., et al. (2005) Erlotinib in previously treated non-small-cell lung cancer, The New England Journal of Medicine, 353, 123-132.
Shi, Y. (2002) Mechanisms of caspase activation and inhibition during apoptosis, Molecular Cell, 9, 459-470.
Sorte, K., et al. (2011) Quantitative assessment of DNA damage directly in lens epithelial cells from senile cataract patients, Molecular Vision, 17, 1-6.
Soussi, T. and Beroud, C. (2001) Assessing TP53 status in human tumours to evaluate clinical outcome, Nature Reviews Cancer, 1, 233-240.
Spigel, D.R., et al. (2011) A phase II study of higher dose weekly topotecan in relapsed small-cell lung cancer, Clinical Lung Cancer, 12, 187-191.
Strasser, A., et al. (1995) Bcl-2 and Fas/APO-1 regulate distinct pathways to lymphocyte apoptosis, The EMBO Journal, 14, 6136-6147.
Strasser, A., O'Connor, L. and Dixit, V.M. (2000) Apoptosis signaling, Annual Review of Biochemistry, 69, 217-245.
Stratton, M.R., Campbell, P.J. and Futreal, P.A. (2009) The cancer genome, Nature, 458, 719-724.
Suzuki, K. and Matsubara, H. (2011) Recent advances in p53 research and cancer treatment, Journal of Biomedicine & Biotechnology, 2011, 978312.
Suzuki, M., Youle, R.J. and Tjandra, N. (2000) Structure of Bax: coregulation of dimer formation and intracellular localization, Cell, 103, 645-654.
Swennen, E.L., et al. (2010) ATP sensitizes H460 lung carcinoma cells to cisplatin-induced apoptosis, Chemico-Biological Interactions, 184, 338-345.
Takaki, T., et al. (2008) Polo-like kinase 1 reaches beyond mitosis--cytokinesis, DNA damage response, and development, Current Opinion in Cell Biology, 20, 650-660.
Taylor, W.R. and Stark, G.R. (2001) Regulation of the G2/M transition by p53, Oncogene, 20, 1803-1815.
Van de Craen, M., et al. (1998) Identification of a new caspase homologue: caspase-14, Cell Death and Differentiation, 5, 838-846.
van Vugt, M.A. and Yaffe, M.B. (2010) Cell cycle re-entry mechanisms after DNA damage checkpoints: giving it some gas to shut off the breaks!, Cell Cycle, 9, 2097-2101.
Videira, M., Almeida, A.J. and Fabra, A. (2011) Preclinical evaluation of a pulmonary delivered paclitaxel-loaded lipid nanocarrier antitumor effect, Nanomedicine : Nanotechnology, Biology, and Medicine, 1-8.
Wajed, S.A., Laird, P.W. and DeMeester, T.R. (2001) DNA methylation: an alternative pathway to cancer, Annals of Surgery, 234, 10-20.
Wang, F., et al. (2003) Taxol inhibits melanoma metastases through apoptosis induction, angiogenesis inhibition, and restoration of E-cadherin and nm23 expression, Journal of Pharmacological Sciences, 93, 197-203.
Wu, F.Y.-H. and Pecq, J.-B.L. (1982) Mechanistic studies of a novel antitumor drug, α-1,3,5- Triglycidyl-s- Triazinetrion, Molecular Pharmacology, 23, 182-189.
Wu, H.C., Chang, D.K. and Huang, C.T. (2006) Targeted therapy for cancer, Journal of Cancer Molecules 2, 57-66.
Yamada, M., et al. (2001) Effect of microsome-liposome fusion on the rotational mobility of cytochrome P450IIB4 in rabbit liver microsomes, Journal of Inorganic Biochemistry, 83, 261-268.
Yang, S., et al. (2003) Triptolide inhibits the growth and metastasis of solid tumors, Molecular Cancer Therapeutics, 2, 65-72.
Youle, R.J. and Strasser, A. (2008) The BCL-2 protein family: opposing activities that mediate cell death, Nature Reviews. Molecular Cell Biology, 9, 47-59.
Yvon, A.M., Wadsworth, P. and Jordan, M.A. (1999) Taxol suppresses dynamics of individual microtubules in living human tumor cells, Molecular Biology of The Cell, 10, 947-959.
Zaffaroni, N., et al. (1998) Induction of apoptosis by taxol and cisplatin and effect on cell cycle-related proteins in cisplatin-sensitive and -resistant human ovarian cells, British Journal Cancer, 77, 1378-1385.
Zou, S., et al. (2011) Identification of dAven, a Drosophila melanogaster ortholog of the cell cycle regulator Aven, Cell Cycle, 10, 989-998.
行政院衛生署 (2012) 100年國人主要死因統計. http://www.doh.gov.tw/CHT2006/DM/DM2_p01.aspx?class_no=25&level_no=1&doc_no=84788.
婦癌基金會 (2012) 台灣100年10大癌症死因. http://www.femalecancer.org.tw/index.php?option=com_content&task=view&id=483&Itemid=31.
聯合新聞網 (2012) 健康醫藥. http://mag.udn.com/mag/life/itempage.jsp?f_SUB_ID=2778.