Skip navigation

DSpace

機構典藏 DSpace 系統致力於保存各式數位資料(如:文字、圖片、PDF)並使其易於取用。

點此認識 DSpace
DSpace logo
English
中文
  • 瀏覽論文
    • 校院系所
    • 出版年
    • 作者
    • 標題
    • 關鍵字
    • 指導教授
  • 搜尋 TDR
  • 授權 Q&A
    • 我的頁面
    • 接受 E-mail 通知
    • 編輯個人資料
  1. NTU Theses and Dissertations Repository
  2. 工學院
  3. 醫學工程學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45081
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor謝銘鈞
dc.contributor.authorHsiang-Yu Hanen
dc.contributor.author韓享祐zh_TW
dc.date.accessioned2021-06-15T04:03:44Z-
dc.date.available2020-12-31
dc.date.copyright2010-02-24
dc.date.issued2010
dc.date.submitted2010-02-11
dc.identifier.citation1. Weitz J, Koch M, Debus J, Hohler T, Galle PR, Buchler MW: Colorectal cancer. Lancet 2005, 365 (9454):153-165.
2. Wu X, Gu J, Wu TT, Swisher SG, Liao Z, Correa AM, Liu J, Etzel CJ, Amos CI, Huang M et al: Genetic variations in radiation and chemotherapy drug action pathways predict clinical outcomes in esophageal cancer. J Clin Oncol 2006, 24 (23):3789-3798.
3. Pomeroy M, Moriarty M: CLINICAL-SIGNIFICANCE OF CELLULAR-RESISTANCE IN TUMORS TO CYTOTOXIC CHEMOTHERAPY AND RADIOTHERAPY. Cytotechnology 1993, 12 (1-3):385-391.
4. Alvarez M, Paull K, Monks A, Hose C, Lee JS, Weinstein J, Grever M, Bates S, Fojo T: Generation of a drug resistance profile by quantitation of mdr-1/P-glycoprotein in the cell lines of the National Cancer Institute Anticancer Drug Screen. J Clin Invest 1995, 95 (5):2205-2214.
5. Glavinas H, Krajcsi P, Cserepes J, Sarkadi B: The role of ABC transporters in drug resistance, metabolism and toxicity. Curr Drug Deliv 2004, 1 (1):27-42.
6. Sarela AI, Macadam RC, Farmery SM, Markham AF, Guillou PJ: Expression of the antiapoptosis gene, survivin, predicts death from recurrent colorectal carcinoma. Gut 2000, 46 (5):645-650.
7. Gil J, Stembalska A, Pesz KA, Sasiadek MM: Cancer stem cells: the theory and perspectives in cancer therapy. J Appl Genet 2008, 49 (2):193-199.
8. Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, Henkelman RM, Cusimano MD, Dirks PB: Identification of human brain tumour initiating cells. Nature 2004, 432 (7015):396-401.
9. Jordan CT, Guzman ML, Noble M: Cancer Stem Cells. N Engl J Med 2006, 355 (12):1253-1261.
10. Wang JCY, Dick JE: Cancer stem cells: lessons from leukemia. Trends Cell Biol 2005, 15 (9):494-501.
11. Chu P, Clanton DJ, Snipas TS, Lee J, Mitchell E, Nguyen ML, Hare E, Peach RJ: Characterization of a subpopulation of colon cancer cells with stem cell-like properties. Int J Cancer 2009, 124 (6):1312-1321.
12. Dittfeld C, Dietrich A, Peickert S, Hering S, Baumann M, Grade M, Ried T, Kunz-Schughart LA: CD133 expression is not selective for tumor-initiating or radioresistant cell populations in the CRC cell lines HCT-116. Radiother Oncol 2009, 92 (3):353-361.
13. Haraguchi N, Ohkuma M, Sakashita H, Matsuzaki S, Tanaka F, Mimori K, Kamohara Y, Inoue H, Mori M: CD133(+)CD44(+) population efficiently enriches colon cancer initiating cells. Ann Surg Oncol 2008, 15 (10):2927-2933.
14. Ricci-Vitiani L, Fabrizi E, Palio E, De Maria R: Colon cancer stem cells. J Mol Med 2009, 87 (11):1097-1104.
15. Dean M: ABC transporters, drug resistance, and cancer stem cells. J Mammary Gland Biol Neoplasia 2009, 14 (1):3-9.
16. Jin F, Li HS, Zhao L, Wei YJ, Zhang H, Guo YJ, Pang R, Jiang XB, Zhao HY: Expression of anti-apoptotic and multi-drug resistance-associated protein genes in cancer stem cell isolated from TJ905 glioblastoma multiforme cell line. Zhonghua Yi Xue Za Zhi 2008, 88 (33):2312-2316.
17. Jin F, Zhao L, Zhao HY, Guo SG, Feng J, Jiang XB, Zhang SL, Wei YJ, Fu R, Zhao JS: Comparison between cells and cancer stem-like cells isolated from glioblastoma and astrocytoma on resistance-associated protein genes. Neuroscience 2008, 154 (2):541-550.
18. Zhao HJ, Peehl DM: Tumor-Promoting Phenotype of CD90(hi) Prostate Cancer-Associated Fibroblasts. Prostate 2009, 69 (9):991-1000.
19. Lan HA, Zain RB, Saitoh M, Muramatsu Y, Shrestha P, Mori M: Proliferating cell nuclear antigen (PCNA) and p53 in epithelial dysplasia and squamous cell carcinoma of oral mucosa - A marker for poor tumor differentiation, increasing nuclear atypia and invasiveness? Anticancer Res 1996, 16 (5B):3059-3065.
20. Shivji MKK, Kenny MK, Wood RD: PROLIFERATING CELL NUCLEAR ANTIGEN IS REQUIRED FOR DNA EXCISION REPAIR. Cell 1992, 69 (2):367-374.
21. Mayol JF, Loeuillet C, Herodin F, Wion D: Characterisation of normal and cancer stem cells: One experimental paradigm for two kinds of stem cells. Bioessays 2009, 31 (9):993-1001.
22. Li HZ, Yi TB, Wu ZY: Suspension culture combined with chemotherapeutic agents for sorting of breast cancer stem cells. BMC Cancer 2008, 8:135.
23. Gerlier D, Thomasset N: USE OF MTT COLORIMETRIC ASSAY TO MEASURE CELL ACTIVATION. Journal of Immunological Methods 1986, 94 (1-2):57-63.
24. Schmandt T, Meents E, Gossrau G, Gornik V, Okabe S, Brustle O: High-purity lineage selection of embryonic stem cell-derived neurons. Stem Cells Dev 2005, 14 (1):55-64.
25. Gan LS, vanSetten G, Seregard S, Fagerholm P: Proliferating cell nuclear antigen colocalization with corneal epithelial stem cells and involvement in physiological cell turnover. Acta Ophthalmol Scand 1995, 73 (6):491-495.
26. Riganti C, Miraglia E, Viarisio D, Costamagna C, Pescarmona G, Ghigo D, Bosia A: Nitric oxide reverts the resistance to doxorubicin in human colon cancer cells by inhibiting the drug efflux. Cancer Res 2005, 65 (2):516-525.
27. Raymond E, Louvet C, Tournigand C, Coudray AM, Faivre S, De Gramont A, Gespach C: Pemetrexed disodium combined with oxaliplatin, SN38, or 5-fluorouracil, based on the quantitation of drug interactions in human HT29 colon cancer cells. Int J Oncol 2002, 21 (2):361-367.
28. Jansen WJ, Zwart B, Hulscher ST, Giaccone G, Pinedo HM, Boven E: CPT-11 in human colon-cancer cell lines and xenografts: characterization of cellular sensitivity determinants. Int J Cancer 1997, 70 (3):335-340.
29. Nissen E, Pauli G, Vollenbroich D: WST-1 assay--a simple colorimetric method for virus titration. In Vitro Cell Dev Biol Anim 1997, 33 (1):28-29.
30. Tavakoli T, Xu XR, Derby E, Serebryakova Y, Reid Y, Rao MS, Mattson MP, Ma W: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines 13, 16 and BG01V. BMC Cell Biol 2009, 10:15.
31. Zhang F, Yan QG, Yan W, Cheng H, Hui YP, Li Q: Cancer is a disease of unregulated expansion of somatic stem cells resulting from disrupted asymmetric division. Med Hypotheses 2008, 70 (1):208-209.
32. Dalerba P, Dylla SJ, Park IK, Liu R, Wang XH, Cho RW, Hoey T, Gurney A, Huang EH, Simeone DM et al: Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci U S A 2007, 104 (24):10158-10163.
33. Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeulen L, Lovino F, Tripodo C, Russo A, Gulotta G, Medema JP et al: Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell 2007, 1 (4):389-402.
34. Schinkel AH, Jonker JW: Mammalian drug efflux transporters of the ATP binding cassette (ABC) family: an overview. Adv Drug Deliv Rev 2003, 55 (1):3-29.
35. Pavillard V, Agostini C, Richard S, Charasson V, Montaudon D, Robert J: Determinants of the cytotoxicity of irinotecan in two human colorectal tumor cell lines. Cancer Chemother Pharmacol 2002, 49 (4):329-335.
36. Konopleva M, Zhao SR, Xie Z, Segall H, Younes A, Claxton DF, Estrov Z, Kornblau SM, Andreeff M: Apoptosis - Molecules and mechanisms. In: Drug Resistance in Leukemia and Lymphoma Iii. Edited by Kaspers GJL, Pieters R, Veerman AJP, vol. 457. New York: Kluwer Academic / Plenum Publ; 1999: 217-236.
37. Liu GT, Yuan XP, Zeng ZH, Tunici P, Ng HS, Abdulkadir IR, Lu LZ, Irvin D, Black KL, Yu JS: Analysis of gene expression and chemoresistance of CDI33(+) cancer stem cells in glioblastoma. Mol Cancer 2006, 5:12.
38. Bunting KD: ABC transporters as phenotypic markers and functional regulators of stem cells. Stem Cells 2002, 20 (1):11-20.
39. Dean M, Fojo T, Bates S: Tumour stem cells and drug resistance. Nat Rev Cancer 2005, 5 (4):275-284.
40. Leung AYH, Leung JCK, Chan LYY, Ma ESK, Kwan TTF, Lai KN, Meng A, Liang R: Proliferating cell nuclear antigen (PCNA) as a proliferative marker during embryonic and adult zebrafish hematopoiesis. Histochem Cell Biol 2005, 124 (2):105-111.
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45081-
dc.description.abstract傳統治療癌症的策略為大量毒殺癌細胞。然而對於癌症後期的病患,這樣的方式並無法有效治癒。目前,越來越多的證據顯示只有少數特定的腫瘤細胞能形成新的癌組織。這些細胞與正常幹細胞所表現的性質有許多相似之處,例如具有自我更新、分化與不斷的增生的能力,因此被稱為癌症幹細胞。
癌症幹細胞因為具有較強的抗藥及抗細胞凋亡機制,不易被化療藥物殺死,而成為腫瘤復發的源頭。因此,進一步研究癌症幹細胞的抗藥及抗細胞凋亡機制有其必要性。在本篇論文中,我們選擇大腸直腸癌幹細胞做相關研究。
首先,利用流式細胞分選儀,將大腸直腸癌病患腫瘤組織分離出來原代培養細胞,以及大腸直腸癌症細胞株(HT-29)中帶有癌症幹細胞表面抗原的細胞篩選出來。
在細胞毒性測試中,我們驗證了癌症幹細胞對目前臨床化療藥物有比較強的抗藥性。接著比較非癌症幹細胞與癌症幹細胞兩者抗藥性及抗細胞凋亡基因的相對表現量。其中在大腸直腸癌症株幹細胞中,抗藥性基因ABCB1及ABCC3有相較於大腸直腸癌細胞8-21倍的表現。至於在抗細胞凋亡基因BCL-XL及NAIP,在人類大腸直腸癌症幹細胞有相較於大腸直腸癌細胞9-12倍高的相對基因表現。
在原代培養細胞中,癌症幹細胞的抗藥性基因ABCB1的表現量約是非癌症幹細胞52倍。至於抗細胞凋亡基因BCL-XL、NAIP、livin及Survivin,在大腸直腸癌幹細胞約高於大腸直腸癌細胞3-58倍的相對基因表現。由以上研究結果可推論,抑制這些高表現量基因,將可降低大腸直腸癌症幹細胞的抗細胞凋亡能力和對化療藥物的抗性,再結合臨床的標靶治療,或許是更有潛力的大腸直腸癌治療模式。
zh_TW
dc.description.abstractIn recent years, the hypothesis of cancer stem-like cells (CSCs) received striking attention. This hypothesis indicated CSCs possess stem cell properties, the ability of self-renew and differentiate, and proliferate infinitely. Besides, they also have characteristics of chemotherapy-resistance and anti-apoptosis. CSCs are more difficult to eradicate and easy to cause cancer relapse because of these properties. Therefore, the mechanisms for CSCs which can escape from death after therapy merits further study.
Here, colorectal cancer cells (CRCs) were studied by investigating the characteristics of drug-resistance and anti-apoptosis within colon CSCs and non-CSCs. First, colon CSCs and non-CSCs were separated by FACS. The cytotoxicity assay showed that CSCs were more resistant to clinic chemotherapeutic drugs than non-CSCs. Moreover, the result of real-time PCR indicated that the gene expression of CSCs on certain drug resistant and anti-apoptosis was at a higher level than non-CSCs. Therefore, these findings provided practical information to design more effective therapeutic strategy for eradicating CRC.
en
dc.description.provenanceMade available in DSpace on 2021-06-15T04:03:44Z (GMT). No. of bitstreams: 1
ntu-99-R96548058-1.pdf: 800477 bytes, checksum: 0aea669938b5d3b4b6de2da67bcf549e (MD5)
Previous issue date: 2010
en
dc.description.tableofcontents中文摘要 Ι
英文摘要 Π
1.Introduction 1
2.Materials and methods 5
2.1 Materials 6
2.2Flow Cytometry Activated Cell Sorting ( FACSAria cell sorter) with CD133 mAb and CD44 mAb Staining 8
2.3 Sphere culture assay 9
2.4 Soft Agar Assay for Colony formation 10
2.5 Immunocytochemistry and confocal microscopy 10
2.6 Evaluation of cytotoxicity (MTT and WST-1 assay) 11
2.7 real-time PCR 12
2.8 Statistical analysis 15
3. Results 16
3.1 Flow Cytometry Activated Cell Sorting ( FACSAria cell sorter) 16
3.2 The stem cell culture condition 16
3.3 The colony formation assay 16
3.4 The proliferation ability determined by PCNA immunofluorescence staining 17
3.5 The sensitivity to chemotherapy drugs 17
3.6 real-time PCR 18
4. Discussion 19
5. Conclusion 23
6. References 24
7. Appendix 31
dc.language.isoen
dc.subject抗藥性zh_TW
dc.subject流式細胞分選儀zh_TW
dc.subject大腸直腸癌zh_TW
dc.subject癌症幹細胞zh_TW
dc.subject抗細胞凋亡zh_TW
dc.subjectcell sortingen
dc.subjectcancer stem-like cellen
dc.subjectsphere cultureen
dc.subjectanti-apoptosisen
dc.subjectdrug resistanceen
dc.subjectreal-time PCRen
dc.title大腸癌細胞與癌幹細胞之抗藥性及抗細胞凋亡特性之研究zh_TW
dc.titleInvestigation of the disparity of chemoresistance and anti-apoptosis between colon cancer cells and colon cancer stem-like cellsen
dc.typeThesis
dc.date.schoolyear98-1
dc.description.degree碩士
dc.contributor.oralexamcommittee楊台鴻,洪士杰,賴秉杉
dc.subject.keyword大腸直腸癌,癌症幹細胞,抗藥性,抗細胞凋亡,流式細胞分選儀,zh_TW
dc.subject.keywordcancer stem-like cell,real-time PCR,drug resistance,anti-apoptosis,sphere culture,cell sorting,en
dc.relation.page38
dc.rights.note有償授權
dc.date.accepted2010-02-11
dc.contributor.author-college工學院zh_TW
dc.contributor.author-dept醫學工程學研究所zh_TW
顯示於系所單位:醫學工程學研究所

文件中的檔案:
檔案 大小格式 
ntu-99-1.pdf
  未授權公開取用
781.72 kBAdobe PDF
顯示文件簡單紀錄


系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。

社群連結
聯絡資訊
10617臺北市大安區羅斯福路四段1號
No.1 Sec.4, Roosevelt Rd., Taipei, Taiwan, R.O.C. 106
Tel: (02)33662353
Email: ntuetds@ntu.edu.tw
意見箱
相關連結
館藏目錄
國內圖書館整合查詢 MetaCat
臺大學術典藏 NTU Scholars
臺大圖書館數位典藏館
本站聲明
© NTU Library All Rights Reserved