請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23353完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 謝銘鈞 | |
| dc.contributor.author | Chia-Ying Lee | en |
| dc.contributor.author | 李佳穎 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:59:40Z | - |
| dc.date.copyright | 2011-08-24 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-19 | |
| dc.identifier.citation | 1. Gottesman, M.M., T. Fojo, and S.E. Bates, Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer, 2002. 2(1): p. 48-58.
2. King-Jen Chang , W.-H.K., Ming-Yang Wang The Epidemiology of Breast Cancer in Taiwan 中華癌醫會誌 (J. Chinese Oncol. Soc.), 2008. 24(2): p. 85-93. 3. Dean, M., A. Rzhetsky, and R. Allikmets, The human ATP-binding cassette (ABC) transporter superfamily. Genome Res, 2001. 11(7): p. 1156-66. 4. Dean, M., T. Fojo, and S. Bates, Tumour stem cells and drug resistance. Nat Rev Cancer, 2005. 5(4): p. 275-84. 5. Ambudkar, S.V., et al., Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annu Rev Pharmacol Toxicol, 1999. 39: p. 361-98. 6. Shen, D.W., et al., Decreased accumulation of [14C]carboplatin in human cisplatin-resistant cells results from reduced energy-dependent uptake. J Cell Physiol, 2000. 183(1): p. 108-16. 7. Schuetz, E.G., W.T. Beck, and J.D. Schuetz, Modulators and substrates of P-glycoprotein and cytochrome P4503A coordinately up-regulate these proteins in human colon carcinoma cells. Mol Pharmacol, 1996. 49(2): p. 311-8. 8. Lowe, S.W., et al., p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell, 1993. 74(6): p. 957-67. 9. Shieh, M.J., et al., Reversal of doxorubicin-resistance by multifunctional nanoparticles in MCF-7/ADR cells. J Control Release, 2011. 152(3): p. 418-25. 10. Zhang, J., et al., Cellular Pharmacokinetic Mechanisms of Adriamycin Resistance and Its Modulation by 20(S)-Ginsenoside Rh2 in MCF-7/Adr Cells. Br J Pharmacol, 2011. 11. Larsen, A.K., A.E. Escargueil, and A. Skladanowski, Resistance mechanisms associated with altered intracellular distribution of anticancer agents. Pharmacol Ther, 2000. 85(3): p. 217-29. 12. Schindler, M., et al., Defective pH regulation of acidic compartments in human breast cancer cells (MCF-7) is normalized in adriamycin-resistant cells (MCF-7adr). Biochemistry, 1996. 35(9): p. 2811-7. 13. Choi, E.W., L.V. Nayak, and P.J. Bates, Cancer-selective antiproliferative activity is a general property of some G-rich oligodeoxynucleotides. Nucleic Acids Res, 2010. 38(5): p. 1623-35. 14. Bates, P.J., et al., Discovery and development of the G-rich oligonucleotide AS1411 as a novel treatment for cancer. Exp Mol Pathol, 2009. 86(3): p. 151-64. 15. Mikami-Terao, Y., et al., Antitumor activity of TMPyP4 interacting G-quadruplex in retinoblastoma cell lines. Exp Eye Res, 2009. 89(2): p. 200-8. 16. Bagalkot, V., et al., An aptamer-doxorubicin physical conjugate as a novel targeted drug-delivery platform. Angew Chem Int Ed Engl, 2006. 45(48): p. 8149-52. 17. Wang, K., et al., Self-assembly of a bifunctional DNA carrier for drug delivery. Angew Chem Int Ed Engl, 2011. 50(27): p. 6098-101. 18. Kim, D., Y.Y. Jeong, and S. Jon, A drug-loaded aptamer-gold nanoparticle bioconjugate for combined CT imaging and therapy of prostate cancer. ACS Nano, 2010. 4(7): p. 3689-96. 19. Huang, Y.F., et al., Molecular assembly of an aptamer-drug conjugate for targeted drug delivery to tumor cells. Chembiochem, 2009. 10(5): p. 862-8. 20. Shieh, Y.A., et al., Aptamer-based tumor-targeted drug delivery for photodynamic therapy. ACS Nano, 2010. 4(3): p. 1433-42. 21. Girvan, A.C., et al., AGRO100 inhibits activation of nuclear factor-kappaB (NF-kappaB) by forming a complex with NF-kappaB essential modulator (NEMO) and nucleolin. Mol Cancer Ther, 2006. 5(7): p. 1790-9. 22. Soundararajan, S., et al., The nucleolin targeting aptamer AS1411 destabilizes Bcl-2 messenger RNA in human breast cancer cells. Cancer Res, 2008. 68(7): p. 2358-65. 23. Otake, Y., et al., Overexpression of nucleolin in chronic lymphocytic leukemia cells induces stabilization of bcl2 mRNA. Blood, 2007. 109(7): p. 3069-75. 24. Chu, T.C., et al., Aptamer mediated siRNA delivery. Nucleic Acids Res, 2006. 34(10): p. e73. 25. Min, K., et al., Dual-aptamer-based delivery vehicle of doxorubicin to both PSMA (+) and PSMA (-) prostate cancers. Biomaterials, 2011. 32(8): p. 2124-32. 26. Manet, I., et al., Affinity of the anthracycline antitumor drugs Doxorubicin and Sabarubicin for human telomeric G-quadruplex structures. Phys Chem Chem Phys, 2011. 13(2): p. 540-51. 27. Buratowski, S. and L.A. Chodosh, Mobility shift DNA-binding assay using gel electrophoresis. Curr Protoc Mol Biol, 2001. Chapter 12: p. Unit 12 2. 28. Liu, Y., F. Wu, and G. Zou, Electrophoresis mobility shift assay and biosensor used in studying the interaction between bleomycin A5 and DNA. Anal Chim Acta, 2007. 599(2): p. 310-4. 29. Trere, D., et al., Qualitative and quantitative analysis of AgNOR proteins in chemically induced rat liver carcinogenesis. Hepatology, 1996. 24(5): p. 1269-73. 30. Pich, A., L. Chiusa, and E. Margaria, Prognostic relevance of AgNORs in tumor pathology. Micron, 2000. 31(2): p. 133-41. 31. Lightner, D.A. and J.E. Gurst, Organic conformational analysis and stereochemistry from circular dichroism spectroscopy. Methods in stereochemical analysis. 2000, New York: Wiley-VCH. xiii, 487 p. 32. David Jones, D.D., Ben Doran, Tomi Odumeru, Kirti Shah, Sanne Kuijper, Colin Green, Nigel Courtenay-Luck, Fiona McLaughlin and Hakim Djeha, AS1411, a novel anti-nucleolin aptamer, shows synergy with other anti-cancer drugs in vitro and in vivo in AML models. 2008. 33. Guedin, A., et al., Sequence effects in single-base loops for quadruplexes. Biochimie, 2008. 90(5): p. 686-96. 34. De Cian, A., et al., Targeting telomeres and telomerase. Biochimie, 2008. 90(1): p. 131-55. 35. Monchaud, D., et al., Ligands playing musical chairs with G-quadruplex DNA: a rapid and simple displacement assay for identifying selective G-quadruplex binders. Biochimie, 2008. 90(8): p. 1207-23. 36. Bao, H., et al., Physical and spectral characterization of the human cyclin A gene and its interactions with anthracycline anticancer drugs. Chemical Physics Letters, 2007. 436(1-3): p. 252-257. 37. Esteve, P.O., H.G. Chin, and S. Pradhan, Molecular mechanisms of transactivation and doxorubicin-mediated repression of survivin gene in cancer cells. J Biol Chem, 2007. 282(4): p. 2615-25. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23353 | - |
| dc.description.abstract | 在本研究中，為了解決多重抗藥性細胞對於化學治療藥物的不敏感性，我們利用人工合成的核酸適體做為同時攜帶化療藥物艾黴素 (doxorubicin-HCl)以及可抑制抗藥性基因表現之siRNA,進行合併治療的載體設計．
第一部分進行核酸適體是否能夠作為艾黴素藥物載體的探討，透過此complex物性的探討以及後續細胞毒殺能力以及細胞內藥物分布實驗的分析，我們發現未進行改殖的AS1411核酸適體並非艾黴素此種藥物的優良載體，必須要後續的改殖及條件調整，才可能改善兩者間鍵結的穩定，並使doxorubicin藥物可透過AS1411核酸適體專一性辨認核仁素的特性，逃脫藥物卡在核內體(endosomes)以及溶酶體(lysosomes)的宿命，使藥物能夠進入抗藥性細胞的細胞核中，達到毒殺能力． 第二部分則是利用卵白素 (streptavidin）以及生物素 (biotin)高度且專一的鍵結能力,將AS1411 核酸適體以及抑制抗藥性基因ADR1表現之siRNA均標定生物素，利用與卵白素結合後形成conjugates,以此conjugates作為抑制抗藥性細胞基因表現的新工具，由流式細胞儀分析的結果，證明此研究中所設計之conjugate的確具有抑制抗藥性基因的效果． | zh_TW |
| dc.description.abstract | To overcome the drug resistance ability of MCF-7/ADR cells, a conjugate containg drug and siRNA was designed. AS1411 aptamer was used as the drug carrier and targeting ligand that specifically bind to nucleolin expressing MCF-7/ADR breast cancer cells. Moreover, the streptavidin-biotin system was used to construct the AS1411/siRNA/ streptavidin conjugate.
Results showed that AS1411 aptamer is not a suitable carrier for doxorubicin drug, or more modifications should be done to make AS1411 aptamer a good doxorubicin carrier. Fourtunately, the silencing ability of AS1411aptamer/ streptavidin/ anti- ABCB1 siRNA conjugate was proved to work, about 50% p-glycoprotein expression was silenced by the siRNA delivered by the conjugate. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:59:40Z (GMT). No. of bitstreams: 1 ntu-100-R98548008-1.pdf: 4785108 bytes, checksum: cd26c5f94c314a6fd687b43ce52d95a7 (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 口試委員會審定書-----------------------------------------------------I
中文摘要------------------------------------------------------------II 英文摘要-----------------------------------------------------------III 目錄----------------------------------------------------------------IV 圖目錄--------------------------------------------------------------VI 表目錄------------------------------------------------------------VIII 1. Introduction--------------------------------------------------------------------------------------1 2. Materials-----------------------------------------------------------------------------------------8 3. Methods-----------------------------------------------------------------------------------------14 3.1Synthesis and Characteristics of the Apt-Doxorubicin Hydrochloride Complex- 15 3.2 Biological Effects of the Apt-Doxorubicin Hydrochloride Complex --------------16 3.3 Synthesis and Characteristics of the AS1411 aptamer: siRNA Complex ---------18 3.4 Biological Effects of AS1411 aptamer: siRNA complex Conjugate ---------------19 3.5 Biological Effects of AS1411 aptamer: siRNA: doxo Complex--------------------20 4. Results-------------------------------------------------------------------------------------------22 4.1 Synthesis and Characteristics of the Apt-Doxorubicin Hydrochloride Complex-22 4.2 Biological Effects of the Apt-Doxorubicin Hydrochloride Complex --------------24 4.3 Synthesis and Characteristics of the AS1411 aptamer: siRNA Complex ---------25 4.4 Biological Effects of AS1411 aptamer: siRNA Complex conjugates Transfection Efficiency Analysis------------------------------------------------------------------------26 4.5 Biological Effects of AS1411 aptamer: siRNA: doxo Complex--------------------26 5. Discussion--------------------------------------------------------------------------------------28 6. Conclusions------------------------------------------------------------------------------------35 7. Future Works---------------------------------------------------------------------------------- 36 8.參考文獻---------------------------------------------------------------------------------------38 9. 附錄-------------------------------------------------------------------------------------------- 43 圖目錄 Fig.1 mechanisms resulting in cellular drug resistance -------------------------------------43 Fig. 2 Fluorescence spectra of doxorubicin solution with increasing molar ratios of the AS1411 aptamer----------------------------------------------------------------------------------45 Fig. 3 Circular Dichroism (CD) spectra-------------------------------------------------------46 Fig. 4 In vitro release profile of free doxorubicin and apt-doxo complex----------------48 Fig.5 Cytotoxicity Effect of free doxorubicin and apt-doxo on MCF-7 cells ------------------------------------------------------------------------------------------------------49 Fig.6 Cytotoxicity Effect of free doxorubicin and apt-doxo on MCF-7/ADR cells ------------------------------------------------------------------------------------------------------50 Fig.7 Subcellular localization of doxorubicin hydrochloride drug (red) in MCF-7/ADR cells-------------------------------------------------------------------------------------------------52 Fig.8 EMSA gel shift analysis of siRNA/aptamer/streptavidin conjugates---------------54 Fig.9 EMSA gel shift analysis of siRNA//aptamer/streptavidin conjugates with reducing agent------------------------------------------------------------------------------------------------55 Fig.10 Comparison of P-glycoprotein expression of MCF-7/ ADR and MCF-7 cells after siRNA transfection by the AS1411aptamer /streptavidin/siRNA conjugate -------------------------------------------------------------------------------------------------------56 Fig.11 Transfection Efficiency and inhibition ability of the AS1411aptamer /streptavidin/siRNA conjugate on the P-glycoprotein expression of MCF-7/ADR cells -------------------------------------------------------------------------------------------------------58 Fig.12 Cytotoxicity Effect of different amount of free doxorubicin and apt-doxo-siRNA complex with different treating time (24 hours, 48 hours, and 72 hours) on MCF-7ADR cells-------------------------------------------------------------------------------------------------61 表目錄 Tabel 1 ABCB binding protein----------------------------------------------------------------43 (modified from http://www.medscape.com/viewarticle/502815) | |
| dc.language.iso | en | |
| dc.subject | 核酸適體 | zh_TW |
| dc.subject | 小干擾核甘核酸 | zh_TW |
| dc.subject | 生物素 | zh_TW |
| dc.subject | 卵白素 | zh_TW |
| dc.subject | 艾黴素 | zh_TW |
| dc.subject | 多重抗藥性 | zh_TW |
| dc.subject | 基因治療 | zh_TW |
| dc.subject | siRNA | en |
| dc.subject | gene therapy | en |
| dc.subject | aptamer | en |
| dc.subject | multidrug resistance | en |
| dc.subject | doxorubicin-HCl | en |
| dc.subject | streptavidin | en |
| dc.subject | biotin | en |
| dc.title | 利用核酸適體作為基因治療及藥物載體抵抗癌症細胞抗藥性之研究 | zh_TW |
| dc.title | Reversal of Multidrug Resistance by Aptamer-Mediated siRNA and Drug Delivery System | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 賴秉杉,羅彩月,婁培人,蕭仲凱 | |
| dc.subject.keyword | 基因治療,核酸適體,多重抗藥性,艾黴素,卵白素,生物素,小干擾核甘核酸, | zh_TW |
| dc.subject.keyword | gene therapy,aptamer,multidrug resistance,doxorubicin-HCl,streptavidin,biotin,siRNA, | en |
| dc.relation.page | 61 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2011-08-19 | |
| 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-100-1.pdf 未授權公開取用 | 4.67 MB | Adobe PDF |
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。