臺灣博碩士論文加值系統

由本研究合成之生物可分解高分子聚乙二醇-聚己內酯-聚乙二醇 (PEG-PCL-PEG) 團聯共聚物形成的奈米微胞，經過動物體外溶血測試得到濃度 2.0 mg/mL以下的奈米微胞溶液具有良好的血液相容性。以未包覆藥物之微胞於動物體內做 14 天急性毒性測試，注射劑量為 71.43 mg/kg，觀察體重變化並沒有發現老鼠體重持續下降的情況，且老鼠存活率為 100 %。進而觀察老鼠的肝臟與腎臟組織切片，其臟器的組織細胞型態屬於正常的情形。kupffer 細胞與 mesangial 細胞分別為肝臟與腎臟裡的發炎細胞，在組織切片中並沒有明顯增多的現象。在單一劑量腫瘤抑制實驗中，注射 doxorubicin 的老鼠在一週內有延緩腫瘤生長情形，注射微胞包覆 doxorubicin 的老鼠在一週後至兩週內的期間有延緩腫瘤生長的情形，並且能降低doxorubicin對肝臟的傷害。而多重劑量腫瘤抑制實驗中，注射微胞包覆 doxorubicin 的腫瘤比注射 doxorubicin 的腫瘤小約 25 %。再經由生物分佈的結果顯示，微胞包覆 doxorubicin 能夠延長在動物體內的血中濃度，由以上結果顯示，生物可分解高分子 PEG-PCL-PEG 具有良好的生物相容性並能搭載藥物延緩腫瘤生長。

In this study, the biodegradable poly(ethylene glycol)-poly(ε-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG) for the nano-sized micelles synthesized. The in vitro hemolysis test showed that the micelles which concentrations of 2.0 mg/mL or less had good blood biocompatibility. The in vitro experiment of acute toxicity using empty micelles (placebo) at dosage of 71.43 mg/kg showed no apparent change of body weight in mice, and the survival rate was 100 %. Furthermore, in the tissue slices of liver and kidney, the cell morphology was normal. And there was unobvious increasing of kupffer cells and mesangial cells in the liver and kidney respectively. In the tumor inhibition experiment, the mice injected with free doxorubicin (DOX) exhibited the delayed tumor growth delay in the first week of administration of the micelle, whereas delayed growth of tumor was found between the first and the second week for mice injected with DOX-loaded PEG-PCL-PEG micelle. In addition, DOX-loaded micelle can reduce the injury to the liver. In the experiment of biodistribution, DOX-loaded micelle could prolong the retention time of DOX in blood. Base on these results, the biodegradable PEG-PCL-PEG triblock copolymer is potential in inhibiting the human breast cancerous tumor growth in nude mice.

目錄
摘要 I
Abstract II
目錄 III
圖索引 VI
表索引 VIII
中英對照表 IX
第一章 緒論 1
1.1 研究背景 1
1.2 研究動機與目的 2
第二章 文獻回顧 3
2.1 抗癌藥物阿黴素 3
2.2 藥物載體 5
2.3 藥物劑量與忍受度 9
2.4 藥物載體進入動物體內的作用 11
第三章 材料與方法 12
3.1實驗架構 12
3.1.1實驗儀器 13
3.1.2實驗藥品 13
3.2 實驗藥品配置 17
3.3 PEG5000 - PCL22000 - PEG5000 (E50C220E50) 材料合成與微胞製作 19
3.3.1 未包覆藥物之微胞製作 19
3.3.2 包覆藥物之微胞製作 19
3.4 溶血測試 21
3.4.1 建立血紅素標準曲線 21
3.4.2 全血定量與調整 21
3.4.3 溶血試驗 21
3.5 E50C220E50 之動物體內毒性測試 23
3.5.1 急性毒性測試 23
3.5.2 白血球計數 24
3.5.3 存活率 24
3.5.4 組織切片觀察 24
3.6 乳癌腫瘤治療評估 26
3.6.1 人類乳癌細胞株 (MCF-7/wt) 培養 26
3.6.2 裸鼠飼養 26
3.6.3 裸鼠腫瘤模型 26
3.6.4 藥物給予 27
3.7 藥物動力學 28
3.7.1 建立 DOX 標準曲線 28
3.7.2 比較液相萃取與固相萃取法 28
3.7.3 動物體內生物分佈 28
第四章 結果與討論 30
4.1 E50C220E50材料特性鑑定 30
4.2 溶血測試結果 31
4.3 動物體內毒性測試 32
4.3.1 體重與白血球計數 32
4.3.2 存活率 34
4.3.3 組織切片觀察 35
4.4 裸鼠腫瘤抑制實驗 (單一劑量) 39
4.4.1 腫瘤模型 40
4.4.2 腫瘤抑制效果 41
4.4.3 裸鼠組織切片觀察 50
4.5 裸鼠腫瘤抑制實驗 (多重劑量) 61
4.5.1 腫瘤抑制效果 61
4.5.2 組織切片觀察 63
4.6 藥物動力學 69
4.6.1 HPLC 條件選定 69
4.6.2 動物體內生物分佈結果 69
第五章 結論 71
參考文獻 72
附件 80
圖索引
圖2 - 1 阿黴素 (DOX) 的化學結構 4
圖2 - 2 微胞 (miccelle) 結構 7
圖2 - 3 脂質體 (liposome) 結構 7
圖2 - 4 樹枝狀高分子 (dendrimer) 結構 8
圖2 - 5 奈米碳管 (carbon nanotube) 結構 8
圖2 - 6 微胞藉由EPR效應從血管到達腫瘤細胞處 11
圖3 - 1 PEG-PCL-PEG 之合成計畫 20
圖3 - 2 溶血測試實驗流程 22
圖3 - 3 血球計數盤 24
圖3 - 4 動物體內生物分佈實驗流程 29
圖4 - 1 E50C220E50 之 FTIR 圖譜分析 30
圖4 - 2 E50C220E50 溶血測試結果 31
圖4 - 3 小鼠毒性測試 0 - 14 天體重記錄 32
圖4 - 4 小鼠毒性測試 0 - 14 天體重記錄，注射高劑量 (92 mg/kg) 控制組 33
圖4 - 5 小鼠毒性測試0 - 14天白血球數量記錄 33
圖4 - 6 小鼠毒性測試0 - 14天老鼠存活率 34
圖4 - 7 毒性測試肝臟組織切片 (HE stain) 36
圖4 - 8 毒性測試肝臟組織切片 (HE stain) 37
圖4 - 9 毒性測試單一高劑量組 (92 mg/kg) 38
圖4 - 10 裸鼠體重記錄 39
圖4 - 11 裸鼠腫瘤生長記錄 41
圖4 - 12 裸鼠1 - L2、2 - non的腫瘤生長趨勢 42
圖4 - 13 裸鼠1 - L2、2 - non的體重變化趨勢 42
圖4 - 14 裸鼠1 - R1、2 - L2 的腫瘤生長趨勢 44
圖4 - 15 裸鼠 1 - R1、2 - L2 的體重變化趨勢 44
圖4 - 16 裸鼠 1 - L1、2 - R1 的腫瘤生長趨勢 45
圖4 - 17 裸鼠 1 - L1、2 - R1 的體重變化趨勢 45
圖4 - 18 裸鼠腫瘤照片(2-R1) 46
圖4 - 19 注射 DOX 溶液的裸鼠腫瘤生長趨勢 48
圖4 - 20 注射DOX溶液後的裸鼠體重變化趨勢 48
圖4 - 21 注射微胞包覆 DOX 溶液的裸鼠腫瘤生長趨勢 49
圖4 - 22 注射微胞包覆DOX溶液後的裸鼠體重變化趨勢 49
圖4 - 23 裸鼠1 - L2腫瘤組織切片(HE stain) 51
圖4 - 24 裸鼠2- non腫瘤組織切片(HE stain) 52
圖4 - 25 裸鼠 1 - R1 腫瘤組織切片(HE stain) 54
圖4 - 26 裸鼠 1 - L1 腫瘤組織切片(HE stain) 55
圖4 - 27 裸鼠 2 - L2 腫瘤組織切片(HE stain) 56
圖4 - 28 裸鼠肝臟組織切片(HE stain) 58
圖4 - 29 裸鼠腎臟組織切片(HE stain) 59
圖4 - 30 裸鼠心臟組織切片(HE stain) 60
圖4 - 31 多重劑量之裸鼠腫瘤生長趨勢 62
圖4 - 32 多重劑量之裸鼠體重變化記錄 62
圖4 - 33 多重劑量裸鼠腫瘤組織切片(HE stain) 64
圖4 - 34 多重劑量裸鼠肝臟組織切片(HE stain) 66
圖4 - 35 多重劑量裸鼠腎臟組織切片(HE stain) 67
圖4 - 36 多重劑量裸鼠心臟組織切片(HE stain) 68
圖4 - 37 注射 DOX 溶液之生物分佈結果 70
圖4 - 38 注射微胞包覆 DOX 溶液之生物分佈結果 70
表索引
表3 - 1 儀器設備目錄 13
表3 - 2 實驗藥品目錄 13

參考文獻
[1] 任一凡，犬乳腺與其腫瘤碳酸酐酶定位及基因表現之研究，國立嘉義大學碩士論文，2006。
[2] Jang SH, Wientjes MG, Lu D, Au JLS. Drug delivery and transport to solid tumors. Pharm Res-Dord. 2003;20:1337-50.
[3] Haley B, Frenkel E. Nanoparticles for drug delivery in cancer treatment. Urol Oncol. 2008;26:57-64.
[4] Singh R, Lillard JW, Jr. Nanoparticle-based targeted drug delivery. Exp Mol Pathol. 2009;86:215-23.
[5] Savic R, Eisenberg A, Maysinger D. Block copolymer micelles as delivery vehicles of hydrophobic drugs: Micelle-cell interactions. Journal of Drug Targeting. 2006;14:343-55.
[6] Zambetti M, Moliterni A, Materazzo C, Stefanelli M, Cipriani S, Valagussa P, et al. Long-term cardiac sequelae in operable breast cancer patients given adjuvant chemotherapy with or without doxorubicin and breast irradiation. J Clin Oncol. 2001;19:37-43.
[7] Wei GL, Xiao SH, Si DY, Liu CX. Improved HPLC method for doxorubicin quantification in rat plasma to study the pharmacokinetics of micelle-encapsulated and liposome-encapsulated doxorubicin formulations. Biomed Chromatogr. 2008;22:1252-8.
[8] Zhang J, Chen XG, Li YY, Liu CS. Self-assembled nanoparticles based on hydrophobically modified chitosan as carriers for doxorubicin. Nanomed-Nanotechnol. 2007;3:258-65.
[9] Letchford K, Burt H. A review of the formation and classification of amphiphilic block copolymer nanoparticulate structures: micelles, nanospheres, nanocapsules and polymersomes. Eur J Pharm Biopharm. 2007;65:259-69.
[10] Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J Clin. 2009;59:225-49.
[11] Jemal A, Siegel R, Ward E, Hao YP, Xu JQ, Murray T, et al. Cancer statistics, 2008. Ca-Cancer J Clin. 2008;58:71-96.
[12] Chung HC, Rha SY, Kim JH, Roh JK, Min JS, Lee KS, et al. P-glycoprotein: the intermediate end point of drug response to induction chemotherapy in locally advanced breast cancer. Breast Cancer Res Treat. 1997;42:65-72.
[13] Nguyen-Van Cuong M-FH, Yung-Tsung Chen, Ian Liau. Synthesis and Characterization of PEG–PCL–PEG Triblock Copolymers as Carriers of Doxorubicin for the Treatment of Breast Cancer. J Appl Polym Sci. 2010;117:3694–703.
[14] Al-Abd AM, Kim NH, Song SC, Lee SJ, Kuh HJ. A simple HPLC method for doxorubicin in plasma and tissues of nude mice. Arch Pharm Res. 2009;32:605-11.
[15] Al-Abd AM, Hong KY, Song SC, Kuh HJ. Pharmacokinetics of doxorubicin after intratumoral injection using a thermosensitive hydrogel in tumor-bearing mice. Journal of Controlled Release. 2010;142:101-7.
[16] Binaschi M, Bigioni M, Cipollone A, Rossi C, Goso C, Maggi CA, et al. Anthracyclines: selected new developments. Curr Med Chem Anticancer Agents. 2001;1:113-30.
[17] Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L. Anthracyclines: Molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev. 2004;56:185-229.
[18] Peters JH, Gordon GR, Kashiwase D, Acton EM. Tissue distribution of doxorubicin and doxorubicinol in rats receiving multiple doses of doxorubicin. Cancer Chemother Pharmacol. 1981;7:65-9.
[19] O'Brien MER, Wigler N, Inbar M, Rosso R, Grischke E, Santoro A, et al. Reduced cardiotoxicity and comparable efficacy in a phase III trial of pegylated liposomal doxorubicin HCl (CAELYX (TM)/Doxil (R)) versus conventional doxorubicin for first-line treatment of metastatic breast cancer. Ann Oncol. 2004;15:440-9.
[20] Biganzoli L, Cufer T, Bruning P, Coleman RE, Duchateau L, Rapoport B, et al. Doxorubicin-paclitaxel - A safe regimen in terms of cardiac toxicity in metastatic breast carcinoma patients. Results from a European organization for research and treatment of cancer multicenter trial. Cancer. 2003;97:40-5.
[21] Olson RD, Mushlin PS. Doxorubicin cardiotoxicity: analysis of prevailing hypotheses. Faseb J. 1990;4:3076-86.
[22] Licata S, Saponiero A, Mordente A, Minotti G. Doxorubicin metabolism and toxicity in human myocardium: Role of cytoplasmic deglycosidation and carbonyl reduction. Chem Res Toxicol. 2000;13:414-20.
[23] Okuda T, Kawakami S, Higuchi Y, Satoh T, Oka Y, Yokoyama M, et al. Enhanced in vivo antitumor efficacy of fenretinide encapsulated in polymeric micelles. Int J Pharm. 2009;373:100-6.
[24] Kataoka K, Harada A, Nagasaki Y. Block copolymer micelles for drug delivery: design, characterization and biological significance. Adv Drug Deliver Rev. 2001;47:113-31.
[25] Prabu P, Chaudhari AA, Dharmaraj N, Khil MS, Park SY, Kim HY. Preparation, characterization, in-vitro drug release and cellular uptake of poly(caprolactone) grafted dextran copolymeric nanoparticles loaded with anticancer drug. J Biomed Mater Res A. 2009;90A:1128-36.
[26] Kim D, Lee ES, Park K, Kwon IC, Bae YH. Doxorubicin Loaded pH-sensitive Micelle: Antitumoral Efficacy against Ovarian A2780/DOXR Tumor. Pharm Res-Dord. 2008;25:2074-82.
[27] Aliabadi HM, Brocks DR, Lavasanifar A. Polymeric micelles for the solubilization and delivery of cyclosporine A: pharmacokinetics and biodistribution. Biomaterials. 2005;26:7251-9.
[28] Ogawara K, Un K, Tanaka K, Higaki K, Kimura T. In vivo anti-tumor effect of PEG liposomal doxorubicin (DOX) in DOX-resistant tumor-bearing mice: Involvement of cytotoxic effect on vascular endothelial cells. J Control Release. 2009;133:4-10.
[29] Gao ZG, Lee DH, Kim DI, Bae YH. Doxorubicin loaded pH-sensitive micelle targeting acidic extracellular pH of human ovarian A2780 tumor in mice. Journal of Drug Targeting. 2005;13:391-7.
[30] Allen C, Maysinger D, Eisenberg A. Nano-engineering block copolymer aggregates for drug delivery. Colloids and Surfaces B: Biointerfaces. 1999;16:3-27.
[31] Danhier F, Magotteaux N, Ucakar B, Lecouturier N, Brewster M, Preat V. Novel self-assembling PEG-p-(CL-co-TMC) polymeric micelles as safe and effective delivery system for Paclitaxel. Eur J Pharm Biopharm. 2009;73:230-8.
[32] Sakakibara T, Chen FA, Kida H, Kunieda K, Cuenca RE, Martin FJ, et al. Doxorubicin encapsulated in sterically stabilized liposomes is superior to free drug or drug-containing conventional liposomes at suppressing growth and metastases of human lung tumor xenografts. Cancer Res. 1996;56:3743-6.
[33] Lasic DD. Doxorubicin in sterically stabilized liposomes. Nature. 1996;380:561-2.
[34] Rawat M, Singh D, Saraf S, Saraf S. Nanocarriers: Promising vehicle for bioactive drugs. Biol Pharm Bull. 2006;29:1790-8.
[35] Ishida T, Shiraga E, Kiwada H. Synergistic antitumor activity of metronomic dosing of cyclophosphamide in combination with doxorubicin-containing PEGylated liposomes in a murine solid tumor model. Journal of Controlled Release. 2009;134:194-200.
[36] Yan W, Huang L. Recent advances in liposome-based nanoparticles for antigen delivery. Polym Rev. 2007;47:329-44.
[37] Hughes GA. Nanostructure-mediated drug delivery. Dm-Dis Mon. 2005;51:342-61.
[38] Aulenta F, Hayes W, Rannard S. Dendrimers: a new class of nanoscopic containers and delivery devices. Eur Polym J. 2003;39:1741-71.
[39] Bianco A, Kostarelos K, Prato M. Applications of carbon nanotubes in drug delivery. Curr Opin Chem Biol. 2005;9:674-9.
[40] Liu Z, Davis C, Cai WB, He L, Chen XY, Dai HJ. Circulation and long-term fate of functionalized, biocompatible single-walled carbon nanotubes in mice probed by Raman spectroscopy. P Natl Acad Sci USA. 2008;105:1410-5.
[41] Zhang XK, Meng LJ, Lu QG, Fei ZF, Dyson PJ. Targeted delivery and controlled release of doxorubicin to cancer cells using modified single wall carbon nanotubes. Biomaterials. 2009;30:6041-7.
[42] Gong CY, Wei XW, Wang XH, Wang YJ, Guo G, Mao YQ, et al. Biodegradable self-assembled PEG-PCL-PEG micelles for hydrophobic honokiol delivery: I. Preparation and characterization. Nanotechnology. 2010;21:-.
[43] Priyabrata P, Tathagata R. Improving liposome integrity and easing bottlenecks to production. Pharmaceutical Technology Europe. 2009;22.
[44] Biricova V, Laznickova A. Dendrimers: Analytical characterization and applications. Bioorg Chem. 2009;37:185-92.
[45] Venkatesan N, Yoshimitsu J, Ito Y, Shibata N, Takada K. Liquid filled nanoparticles as a drug delivery tool for protein therapeutics. Biomaterials. 2005;26:7154-63.
[46] Park K, Lee GY, Kim YS, Yu M, Park RW, Kim IS, et al. Heparin-deoxycholic acid chemical conjugate as an anticancer drug carrier and its antitumor activity. Journal of Controlled Release. 2006;114:300-6.
[47] Liu BR, Yang M, Li RT, Ding YT, Qian XP, Yu LX, et al. The antitumor effect of novel docetaxel-loaded thermosensitive micelles. Eur J Pharm Biopharm. 2008;69:527-34.
[48] Son YJ, Jang JS, Cho YW, Chung H, Park RW, Kwon IC, et al. Biodistribution and anti-tumor efficacy of doxorubicin loaded glycol-chitosan nanoaggregates by EPR effect. Journal of Controlled Release. 2003;91:135-45.
[49] Lee ES, Na K, Bae YH. Doxorubicin loaded pH-sensitive polymeric micelles for reversal of resistant MCF-7 tumor. Journal of Controlled Release. 2005;103:405-18.
[50] Laginha KM, Verwoert S, Charrois GJR, Allen TM. Determination of doxorubicin levels in whole tumor and tumor nuclei in murine breast cancer tumors. Clin Cancer Res. 2005;11:6944-9.
[51] Wang X, Wang YG, Chen XM, Wang JC, Zhang X, Zhang Q. NGR-modified micelles enhance their interaction with CD13-overexpressing tumor and endothelial cells. Journal of Controlled Release. 2009;139:56-62.
[52] Kim TH, Mount CW, Gombotz WR, Pun SH. The delivery of doxorubicin to 3-D multicellular spheroids and tumors in a murine xenograft model using tumor-penetrating triblock polymeric micelles. Biomaterials. 2010.
[53] Oh KT, Lee ES, Kim D, Bae YH. L-Histidine-based pH-sensitive anticancer drug carrier micelle: Reconstitution and brief evaluation of its systemic toxicity. Int J Pharm. 2008;358:177-83.
[54] Chytil P, Etrych T, Konak C, Sirova M, Mrkvan T, Boucek J, et al. New HPMA copolymer-based drug carriers with covalently bound hydrophobic substituents for solid tumour targeting. Journal of Controlled Release. 2008;127:121-30.
[55] Zhang C, Qu GW, Sun YJ, Wu XJ, Yao ZL, Guo QL, et al. Pharmacokinetics, biodistribution, efficacy and safety of N-octyl-O-sulfate chitosan micelles loaded with paclitaxel. Biomaterials. 2008;29:1233-41.
[56] Vega-Villa KR, Takemoto JK, Yanez JA, Remsberg CM, Forrest ML, Davies NM. Clinical toxicities of nanocarrier systems. Adv Drug Deliver Rev. 2008;60:929-38.
[57] Bae Y, Nishiyama N, Fukushima S, Koyama H, Yasuhiro M, Kataoka K. Preparation and biological characterization of polymeric micelle drug carriers with intracellular pH-triggered drug release property: Tumor permeability, controlled subcellular drug distribution, and enhanced in vivo antitumor efficacy. Bioconjugate Chem. 2005;16:122-30.
[58] Moghimi SM, Hunter AC, Murray JC. Long-circulating and target-specific nanoparticles: Theory to practice. Pharmacol Rev. 2001;53:283-318.
[59] Yokoyama M, Okano T, Sakurai Y, Fukushima S, Okamoto K, Kataoka K. Selective delivery of adriamycin to a solid tumor using a polymeric micelle carrier system. J Drug Target. 1999;7:171-86.
[60] Hamaguchi T, Matsumura Y, Suzuki M, Shimizu K, Goda R, Nakamura I, et al. NK105, a paclitaxel-incorporating micellar nanoparticle formulation, can extend in vivo antitumour activity and reduce the neurotoxicity of paclitaxel. Br J Cancer. 2005;92:1240-6.
[61] Li Y, Qi XR, Maitani Y, Nagai T. PEG-PLA diblock copolymer micelle-like nanoparticles as all-trans-retinoic acid carrier: in vitro and in vivo characterizations. Nanotechnology. 2009;20:5.
[62] Xiong XB, Huang Y, Lu WL, Zhang X, Zhang H, Nagai T, et al. Enhanced intracellular delivery and improved antitumor efficacy of doxorubicin by sterically stabilized liposomes modified with a synthetic RGD mimetic. Journal of Controlled Release. 2005;107:262-75.
[63] Gordon AN, Fleagle JT, Guthrie D, Parkin DE, Gore ME, Lacave AJ. Recurrent epithelial ovarian carcinoma: A randomized phase III study of pegylated liposomal doxorubicin versus topotecan. J Clin Oncol. 2001;19:3312-22.
[64] Muggia FM. Doxorubicin-polymer conjugates: further demonstration of the concept of enhanced permeability and retention. Clin Cancer Res. 1999;5:7-8.
[65] Hoffman AS. The origins and evolution of "controlled" drug delivery systems. J Control Release. 2008;132:153-63.
[66] Longmuir KJ, Haynes SM, Baratta JL, Kasabwalla N, Robertson RT. Liposomal delivery of doxorubicin to hepatocytes in vivo by targeting heparan sulfate. Int J Pharm. 2009;382:222-33.
[67] Saad M, Garbuzenko OB, Ber E, Chandna P, Khandare JJ, Pozharov VP, et al. Receptor targeted polymers, dendrimers, liposomes: Which nanocarrier is the most efficient for tumor-specific treatment and imaging? Journal of Controlled Release. 2008;130:107-14.
[68] Schluep T, Hwang J, Cheng JJ, Heidel JD, Bartlett DW, Hollister B, et al. Preclinical efficacy of the camptothecin-polymer conjugate IT-101 in multiple cancer models. Clin Cancer Res. 2006;12:1606-14.
[69] 陳怡坊，大豆異黃酮對DMBA誘發有無卵巢切除雌鼠乳腺腫瘤之延緩效應，北醫學大學碩士論文，2004。
[70] Lee ES, Gao Z, Kim D, Park K, Kwon IC, Bae YH. Super pH-sensitive multifunctional polymeric micelle for tumor pH(e) specific TAT exposure and multidrug resistance. J Control Release. 2008;129:228-36.
[71] American Society for Test and Materials. ASTM F756-00 2000.
[72] 黃華民，徐淑媛，組織學彩色圖譜，合記圖書，2002。
[73] 徐國成，邱雪杉，韓秋生，病理學彩色圖譜，新文京開發，2005。
[74] Brunner N, Bronzert D, Vindelov LL, Rygaard K, Spang-Thomsen M, Lippman ME. Effect on growth and cell cycle kinetics of estradiol and tamoxifen on MCF-7 human breast cancer cells grown in vitro and in nude mice. Cancer Res. 1989;49:1515-20.