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  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/123456789/22676
完整後設資料紀錄
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dc.contributor.advisor謝銘鈞
dc.contributor.authorHuei-Ru Linen
dc.contributor.author林慧如zh_TW
dc.date.accessioned2021-06-08T04:24:18Z-
dc.date.copyright2011-08-22
dc.date.issued2011
dc.date.submitted2011-08-18
dc.identifier.citation1. The Department of Health, Executive Yuan 2007 Cancer registry annual report. R.O.C., Taiwan.
2. Markowitz SD, Bertagnolli MM: Molecular Basis of Colorectal Cancer. N Engl Med. 2009; 361 (25):2449-60.
3. Fearon ER, Vogelstein B: A genetic model for colorectal tumorigenesis. Cell 1990; 61(5):759-767.
4. Saif MW, Chu E: Biology of colorectal cancer. The Cancer Journal 2010; 16(3):196-201.
5. Weitz J, Koch M, Debus J, Hohler T, Galle PR, Buchler MW: Colorectal Cancer. Lancet 2005; 365(9454):153-165.
6. Compton CC: Colorectal carcinoma: Diagnostic, prognostic, and molecular features. Mod Pathol. 2003; 16(4):376-388.
7. Ruers T, Bleichrodt RP: Treatment of liver metastases, an update on the possibilities and results. Eur J Cancer 2002; 38(7):1023-1033.
8. O’Connell JB, Maggard MA, Ko CY: Colon cancer survival rates with the new American Joint Committee on Cancer sixth edition staging. J Natl Cancer Inst. 2004; 96(19):1420-1425.
9. Kollmar O, Rupertus K, Scheuer C, Nickels RM, Haberl GC, Tilton B, Menger MD, Schilling MK: CXCR4 and CXCR7 regulate angiogenesis and CT26. WT tumor growth independent from SDF-1. Int J Cancer 2010; 126(6):1302-1315.
10. Chung DC: Molecular prognostic markers and colorectal cancer: The search goes on. Gastroenterology 1998; 114(6):1330-1332.
11. Geiger TR, Peeper DS: Metastasis mechanisms. Biochim Biophys Acta. 2009; 1796(2):293-308.
12. Robertson JH, Sarkar S, Yang SY, Seifalian AM, Winslet MC: In vivo models for early development of colorectal liver metastasis. Int J Exp Pathol. 2008; 89(1):1-12.
13. Gil J, Stembalska A, Pesz KA, Sasiadek MM: Cancer stem cells: The theory and persectives in cancer therapy. J Appl Genet 2008; 49(2):193-199.
14. Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, Henkelman RM, Cusimano MD, Dirks PB: Identification of human brain tumor initiating cells. Nature 2004; 432(7015):396-401.
15. Jordan CT, Guzman ML, Noble M: Cancer Stem Cells. N Engl J Med. 2006; 355(12):1253-1261.
16. Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF: From the cover: Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A. 2003; 100(7):3983-3988.
17. Li C, Heidt DG, Dalerba P, Burant CF, Zhang L, Adsay V, Wicha M, Clarke MF, Simeone DM: Identification of pancreatic cancer stem cells. Cancer Res. 2007; 67(3):1030-1037.
18. O’Brien CA, Pollett A, Gallinger A, Dick JE: A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 2007; 445(7123):106-110.
19. 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.
20. Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C, De Maria R: Identification and expansion of human colon-cancer-initiating cells. Nature 2007; 445(7123):111-115.
21. Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, Bruns CJ, Heeschen C: Distinct populations of cancer stem cells determine tumor growth and metaststic activity in human pancreatic cancer. Cell Stem Cell 2007; 1(3):313-323.
22. Davis SJ, Divi V, Owen JH, Bradford CR, Carey TE, Papagerakis S, Prince ME: Metastatic potential of CSCs in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 2010; 136(12):1260-1266.
23. Anderson EC, Hessman C, Levin TG, Monroe MM, Wong MH: The role of colorectal cancer stem cells in metastatic disease and therapeutic response. Cancers 2011; 3(1):319-339.
24. Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T, St Clair R, Baljevic M, White I, Jin DK, Chadburn A, Murphy AJ, Valenzuela DM, Gale NW, Thurston G, Yancopoulos GD, D'Angelica M, Kemeny N, Lyden D, Rafii S: CD133 expression is not restricted to stem cells, and both CD133+ and CD133- metastatic colon cancer cells initiate tumors. J Clin Invest. 2008; 118(6):2111-20.
25. Pang R, Law WL, Chu AC, Poon JT, Lam CS, Chow AK, Ng L, Cheung LW, Lan XR, Lan HY, Tan VP, Yau TC, Poon RT, Wong BC: A subpopulation of CD26+ cancer stem cells with metastatic capacity in human colorectal cancer. Cell Stem Cell. 2010; 6(6):603-15.
26. de Jong GM, Aarts F, Hendriks T, Boerman OC, Bleichrodt RP: Animal models for liver metastases of colorectal cancer: research review of preclinical studies in rodents. J Surg Res. 2009; 154(1):167-76.
27. Leduc EH: Metastasis of transplantable hepatomas from the spleen to the liver in mice. Cancer Res. 1959; 19:1091-1095.
28. Kozlowski JM, Fidler IJ, Campbell D, Xu ZL, Kaighn ME, Hart IR: Metastatic behavior of human tumor cell lines grown in the nude mouse. Cancer Res. 1984; 44(8):3522-3529.
29. Bouvet M, Tsuji K, Yang M, Jiang P, Moossa AR, Hoffman RM: In vivo color-coded imaging of the interaction of colon cancer cells and splenocytes in the formation of liver metastases. Cancer Res. 2006; 66(23):11293-7.
30. Edinger M, Cao YA, Hornig YS, Jenkins DE, Verneris MR, Bachmann MH, Negrin RS, Contag CH: Advancing animal models of neoplasia through in vivo bioluminescence imaging. Eur J Cancer 2002; 38(16):2128-36.
31. Buchhorn HM, Seidl C, Beck R, Saur D, Apostolidis C, Morgenstern A, Schwaiger M, Senekowitsch-Schmidtke R: Non-invasive visualisation of the development of peritoneal carcinomatosis and tumour regression after 213Bi-radioimmunotherapy using bioluminescence imaging. Eur J Nucl Med Mol Imaging 2007; 34(6):841-849.
32. Zeamari S, Rumping G, Floot B, Lyons S, Stewart FA: In vivo bioluminescence imaging of locally disseminated colon carcinoma in rats. Br J Cancer 2004; 90(6):1259-1264.
33. Rehemtulla A, Stegman LD, Cardozo SJ, Gupta S, Hall DE, Contag CH, Ross BD: Rapid and quantitative assessment of cancer treatment response using in vivo bioluminescence imaging. Neoplasia 2000; 2(6):491-495.
34. Saur D, Seidler B, Schneider G, Algül H, Beck R, Senekowitsch-Schmidtke R, Schwaiger M, Schmid RM: CXCR4 expression increases liver and lung metastasis in a mouse model of pancreatic cancer. Gatroenterology 2005; 129(4):1237-1250.
35. Kingston RE, Chen CA, Okayama H: Calcium phosphate transfection. Curr Protoc Cell Biol. 2003; Chapter 20: Unit 20.3.1-20.3.8.
36. 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.
37. Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeulen L, Iovino F, Tripodo C, Russo A, Gulotta G, Medema JP, Stassi G: Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell. 2007; 1 (4):389-402.
38. Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA, and Daidone MG: Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res. 2005; 65 (13):5506-5511.
39. Chen JY, Tang YA, Huang SM, Juan HF, Wu LW, Sun YC, Wang SC, Wu KW, Balraj G, Chang TT, Li WS, Cheng HC, Wang YC: A novel sialyltransferase inhibitor suppresses FAK/paxillin signaling and cancer angiogenesis and metastasis pathways. Cancer Res. 2011; 71(2):473-83.
40. Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM, McEwan RN: A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res. 1987; 47(12):3239-45.
41. Liu L, Zhang Q, Zhang Y, Wang S, Ding Y: Lentivirus-mediated silencing of Tiam1 gene influences multiple functions of a human colorectal cancer cell line. Neoplasia. 2006; 8(11):917-924.
42. Shaw LM: Tumor cell invasion assays. Methods Mol Biol. 2005; 294:97-105.
43. Buchanan FG, Gorden DL, Matta P, Shi Q, Matrisian LM, DuBois RN: Role of beta-arrestin 1 in the metastatic progression of colorectal cancer. Proc Natl Acad Sci USA. 2006; 103(5):1492-1497.
44. Susan JB, Bruce R: Metastatic model of colon carcinoma in mice: Utility in the study of tumor growth and progression. Curr Protoc in Pharmacol. 2007; Chapter 14(38): Unit 14.5.1-14.5.13.
45. Li G, Liu C, Yuan J, Xiao X, Tang N, Hao J, Wang H, Bian X, Deng Y, Ding Y: CD133(+) single cell-derived progenies of colorectal cancer cell line SW480 with different invasive and metastatic potential. Clin Exp Metastasis 2010; 27(7):517-527.
46. Belov L, Zhou J, Christopherson RI: Cell surface markers in colorectal cancer prognosis. Int J Mol Sci. 2010; 12(1):78-113.
47. Balic M, Lin H, Young L, Hawes D, Giuliano A, McNamara G, Datar RH, Cote RJ: Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype. Clin Cancer Res. 2006; 12(19): 5615-5621.
48. Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, Bruns CJ, Heeschen C: Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell. 2007; 1(3):313-323.
49. Rice BW, Cable MD, Nelson MB: In vivo imaging of light-emitting probes. J Biomed Opt. 2001; 6(4):432-440.
50. Strube A, Stepina E, Mumberg D, Scholz A, Hauff P, Käkönen SM: Characterization of a new renal cell carcinoma bone metastasis mouse model. Clin Exp Metastasis. 2010; 27(5):319-30.
51. Zhang H, Qiao H, Bakken A, Gao F, Huang B, Liu YY, El-Deiry W, Ferrari VA, Zhou R: Utility of dual-modality bioluminescence and MRI in monitoring stem cell survival and impact on post myocardial infarct remodeling. Acad Radiol. 2011; 18(1):3-12.
52. Inoue Y, Izawa K, Tojo A, Nomura Y, Sekine R, Oyaizu N, Ohtomo K: Monitoring of disease progression by bioluminescence imaging and magnetic resonance imaging in an animal model of hematologic malignancy. Exp Hematol. 2007; 35(3):407-15.
53. Ribatti D, Mangialardi G, Vacca A: Stephen Paget and the 'seed and soil' theory of metastatic dissemination. Clin Exp Med. 2006; 6(4):145-149.
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22676-
dc.description.abstract癌症轉移是造成癌症病患死亡的重要原因,在罹患大腸直腸癌的患者中很高比例會發生肝轉移的現象,嚴重影響大腸直腸癌患者的預後情形。近年來,許多研究致力於癌症幹細胞的理論,證據顯示腫瘤細胞中只有少數細胞有能力形成新的腫瘤病灶,並且主導腫瘤組織的生長,也有愈來愈多的研究結果指出癌症幹細胞不只與癌症的惡性程度有關,也極有可能在癌症轉移、復發的過程中扮演重要角色。目前,我們對這項假設的詳細機制所知不多,也缺乏完整建立的活體動物模型系統來進行這方面的探討。因此,設計實驗方法研究大腸直腸癌幹細胞在肝轉移過程中所扮演的角色有其必要性。
在本篇論文中,利用細胞試驗和動物試驗,並結合非侵入性的小動物活體影像系統來探討大腸直腸癌幹細胞的轉移潛力。首先,為了進行非侵入性光學影像的觀察,人類大腸直腸癌細胞株皆透過病毒的方式轉染冷光酵素基因。在體外細胞試驗的部份,主要研究細胞的侵襲力和移動能力,在體內動物實驗的部份,注射癌症細胞株和癌症幹細胞進入小鼠脾臟,透過門脈循環散播細胞,進一步結合非侵入性的活體影像系統觀察植入細胞在動物體內的生長及轉移情形。
結果顯示帶有癌症幹細胞表面抗原比例高的細胞株相較於抗原比例低的細胞株可能表現出較強的侵襲力和移動能力,在小鼠體內較易形成肝轉移病灶,但其表現程度與抗原表現程度並未完全一致,進一步利用癌症幹細胞表面抗原篩選出癌症幹細胞,細胞試驗結果發現這些細胞具有較強的侵襲和移動能力,在小鼠體內形成腫瘤的能力也比非癌症幹細胞強。由以上研究結果可知,此種結合體外試驗及動物模型並利用非侵入性活體影像系統的方法,應用於評估細胞轉移情形的研究是可行的,此外大腸直腸癌幹細胞確實表現出較強的轉移潛力。
zh_TW
dc.description.abstractCancer metastasis is the primary cause of cancer-related death. Liver is the major site of metastasis for patient with colorectal cancer and it is also the major determinant of survival. In the last several years, cancer stem cells (CSCs) have been identified in several human malignancies, including colorectal cancer. Moreover, more and more studies evidence that CSCs may not only involve in malignant cancer initiation and progression, but also drive cancer metastasis and responsible for tumor recurrence, result in the poor prognostic. However, there is little known about the role of CSCs in metastasis of human cancer, also lacking a well established in vivo animal model system for present studies. Therefore, it is necessary to help elucidate the role of colorectal cancer stem cells in the liver metastatic spread.
In this study, in vitro cells experiments and in vivo animal models were established to observe colorectal cancer stem-like cells metastasis. In order to visualize by bioluminescence imaging system, cancer cells were all transfected with firefly luciferase gene. The metastatic potential of four colon cancer cell lines and CD133 sorted CSCs were assessed. The results showed that the CD133high cell lines were performed higher metastatic potential than CD133low cell lines, but not exact corresponded with CD133 expression levels. On the other hand, in vitro results indicated that the CD133+ cancer cells have great capacities of invasion and migration than CD133- cancer cells; in vivo experimental results showed that CD133+ cells expressed great tumor initiated ability in comparison with CD133- cells.
In conclusion, the in vitro and in vivo model in this study can give better way to help understand the role of CD133+ colorectal cancer stem-like cells in liver metastasis, and also apply to other type of cancer stem cells metastatic researches. Additionally, the CD133+ cancer cells may play a crucial role in colorectal cancer metastatic process.
en
dc.description.provenanceMade available in DSpace on 2021-06-08T04:24:18Z (GMT). No. of bitstreams: 1
ntu-100-R98548027-1.pdf: 3925481 bytes, checksum: eaf6e06c3301c9027d896cdc180759ce (MD5)
Previous issue date: 2011
en
dc.description.tableofcontents中文摘要 I
Abstract II
1. Introduction 1
1.1 Colorectal cancer and metastasis 1
1.2 Colon cancer stem cells 2
1.3 Experimental liver metastasis animal model 3
1.4 Non-invasive imaging 5
1.5 Aim of study 6
2. Materials and methods 7
2.1 Flowchart 7
2.2 Materials 8
2.3 Cell culture 13
2.4 Luciferase and green fluorescence genes transfection 13
2.4.1 Transduced cells selecting 14
2.4.2 Luciferase activities assay 14
2.5 Flow cytometry analysis for CD133 expression of colon cancer cell lines 15
2.6 Cell sorting for isolation of cancer stem cells (CSCs) 16
2.7 Invasion and migration assays 17
2.7.1 Wound-healing cell migration assay 17
2.7.2 Boyden chamber cell invasion assay 18
2.7.3 Transwell cell migration assay 19
2.8 In vivo experimental metastasis assay 19
2.8.1 Animals 19
2.8.2 Intrasplenic injection model for colon cancer liver metastasis 20
2.9 Non-invasive imaging system 21
2.9.1 Bioluminescence imaging 21
2.9.2 High frequency ultrasound imaging 22
2.10 Statistical analysis 23
3. Results 24
3.1 Luciferase activities assay 24
3.2 Flow cytometry analysis for CD133 expression of colon cancer cell lines 24
3.3 Invasion and migration assays 24
3.4 In vivo experimental metastasis assay 25
4. Discussion 28
5. Conclusion 32
6. Reference 34
7. Appendix 43
dc.language.isoen
dc.subject活體影像系統zh_TW
dc.subject癌症幹細胞zh_TW
dc.subject動物模型zh_TW
dc.subject大腸直腸癌zh_TW
dc.subject肝轉移zh_TW
dc.subjectanimal modelen
dc.subjectcolorectal canceren
dc.subjectliver metastasisen
dc.subjectcancer stem cellsen
dc.subjectbioluminescence imagingen
dc.subjectultrasounden
dc.title利用細胞試驗及動物活體影像系統評估人類大腸直腸癌幹細胞之轉移潛力zh_TW
dc.titleUtilizing In vitro and In vivo System to Assess the Metastatic Potential of Cancer Stem-like Cells in Human Colorectal Canceren
dc.typeThesis
dc.date.schoolyear99-2
dc.description.degree碩士
dc.contributor.oralexamcommittee蕭仲凱,婁培人,羅彩月,賴秉杉
dc.subject.keyword肝轉移,癌症幹細胞,活體影像系統,大腸直腸癌,動物模型,zh_TW
dc.subject.keywordcolorectal cancer,liver metastasis,cancer stem cells,bioluminescence imaging,ultrasound,animal model,en
dc.relation.page56
dc.rights.note未授權
dc.date.accepted2011-08-18
dc.contributor.author-college工學院zh_TW
dc.contributor.author-dept醫學工程學研究所zh_TW
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