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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 江俊斌(Chun-Pin Chiang),郭英雄(Ying-Shiung Kuo) | |
dc.contributor.author | Chien-Yang Yeh | en |
dc.contributor.author | 葉建陽 | zh_TW |
dc.date.accessioned | 2021-06-08T05:28:06Z | - |
dc.date.copyright | 2005-08-02 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-07-13 | |
dc.identifier.citation | REFERENCES:
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/24491 | - |
dc.description.abstract | 背景和目的︰以往的研究顯示環氧化酵素-2 (Cyclooxygenase-2, COX-2) 在口腔癌細胞中有過度表現的情形。而COX-2 本身會透過血管生成,細胞增殖及轉移等方式,來促進口腔癌細胞的擴展。5-氨基酮戊酸之光動力治療(ALA-PDT),在針對口腔癌前病變與口腔癌的治療上,已有不錯的療效。Celebrex為一種選擇性的COX-2 抑制劑,目前已被當成一種化學預防性的藥物,應用在不同類型的癌症治療上。在本研究中,我們探討是否Celebrex的給予,對於ALA-PDT在口腔癌細胞株的生長抑制上,能有加強的效果。以及Celebrex的給予,是否能夠增強ALA-PDT所誘導之口腔癌細胞之凋亡。
材料和方法︰在這項實驗中,我們使用兩株不同的口腔癌細胞株,SAS和Ca9-22。將個別SAS和Ca9-22的口腔癌細胞株,分成四個組別,分別以單獨照光、給予Celebrex並照光、ALA-PDT、給予Celebrex合併ALA-PDT處理,以西方墨點法,探討口腔癌細胞株中COX-2的表現;以MTT法,評估口腔癌細胞株活性;以Hoechst 33258螢光染色,研究口腔癌細胞株凋亡細胞的形態;以DNA laddering 分析法,研究口腔癌細胞株凋亡所引起的DNA片斷碎裂等。 結果︰ 在沒有任何外界刺激的情況之下,Ca9-22細胞有COX-2的表現,而SAS細胞則無。當以ALA-PDT處理時,可以發現SAS細胞的COX-2會被誘發而表現,Ca9-22細胞的COX-2則會有增強表現的情形,且COX-2之表現,有劑量與時間依存的情形。若SAS細胞給予Celebrex,再以ALA-PDT處理時,則會減少SAS細胞中COX-2之表現。以MTT細胞活性分析,和未做任何治療的控制組比較,可以發現SAS與Ca9-22細胞,在48小時的培養後,給予Celebrex的組別,會有明顯的細胞生長抑制之情形(p<0.05)。當SAS和Ca9-22細胞在單獨照光和給予Celebrex合併照光之後,並培養4小時的組別裡,縱使光照的能量增強,兩者皆不會有顯著細胞生長抑制的情形。在ALA-PDT和Celebrex合併ALA-PDT的治療組別,則有顯著的細胞生長抑制之情形(p<0.05)。這個協同增強生長抑制的效果,是發生在Celebrex合併ALA-PDT治療的組別上。以Hoechst 33258螢光染色觀察顯示,在光照 (SAS, 40 J/cm2; Ca9-22, 4 J/cm2) 後的72小時,可以發現兩株口腔癌細胞,有ALA-PDT所誘導的細胞凋亡之現象產生。不過更多細胞凋亡的情形,發生在Celebrex合併ALA-PDT的治療組別上。DNA laddering 分析亦顯示Celebrex合併ALA-PDT治療的組別,會比單獨作ALA-PDT治療的組別,有較早產生細胞凋亡、較大量產生細胞凋亡、以及光照劑量較少即有大量細胞凋亡產生之情形。 結論︰ 由於Celebrex的給予,確實能增強ALA-PDT對SAS與Ca9-22口腔癌細胞生長抑制的效果,並且亦能增加ALA-PDT對SAS與Ca9-22所誘導的細胞凋亡,我們認為光動力治療(ALA-PDT)合併COX-2抑制劑治療,對口腔癌細胞株之毒殺上,有協同增強的治療成效。 | zh_TW |
dc.description.abstract | Background and purpose: Previous studies showed overexpression of cyclooxygenase-2 (COX-2) in oral cancers. COX-2 may promote oral cancer progression via an increase in tumor angiogenesis, cell proliferation, and metastasis. 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) has been used to treat oral premalignant and malignant lesions with promising results. A selective COX-2 inhibitor – Celebrex has been used as a chemopreventive drug for different types of cancer. In this study, we tested whether addition of Celebrex to ALA-PDT could increase the inhibitory effect of ALA-PDT on the growth of oral cancer cell lines and could enhance the ALA-PDT-induced apoptosis of oral cancer cell lines.
Materials and methods: Two oral cancer cell lines, SAS and Ca9-22, were treated with light alone, Celebrex plus light, ALA-PDT alone, and Celebrex plus ALA-PDT. Western blot, MTT assay, Hoechst 33258 staining, and DNA laddering analysis were used to study the COX-2 expression, cell viability, morphology of apoptotic cells, and apoptosis-induced DNA fragmentation in SAS and Ca9-22 cells, respectively. Results: The COX-2 expression was found in Ca9-22 cells but not in SAS cells without external stimulation. The COX-2 expression could be induced in SAS cells and up-regulated in Ca9-22 cells by ALA-PDT in a time- and dose-dependent manner. Addition of Celebrex to the cultures decreased the ALA-PDT-induced COX-2 expression in SAS cells. Compared to the control group without any treatment, a significant inhibition of growth of SAS and Ca9-22 cells was found by the MTT assay after co-incubation with Celebrex for 48 hours (P<0.05). When SAS or Ca9-22 cells were treated with a series of increasing light doses and then cultured for 4 hours, compared to the control group no significant inhibition of cell growth was found in the light only and Celebrex plus light groups, whereas a significant inhibition of cell growth was found in ALA-PDT alone and Celebrex plus ALA-PDT groups (P<0.05). A synergic inhibition of cell growth was found when cells were treated with Celebrex plus ALA-PDT. ALA-PDT-induced apoptosis of SAS and Ca9-22 cells could be observed by fluorescence microscopy with the aid of Hoechst 33258 staining 72 hours after treatment with ALA-PDT with the light dose of 40 and 4 J/cm2, respectively. More apoptotic SAS and Ca9-22 cells were found in the Celebrex plus ALA-PDT group than in the ALA-PDT alone group. DNA laddering analysis showed that apoptosis-induced DNA fragmentation occurred earlier, was induced with less light dose, and was more abundant in the Celebrex plus ALA-PDT group than in the ALA-PDT alone group. Conclusion: Because addition of Celebrex to ALA-PDT could increase the inhibitory effect of ALA-PDT on the growth of SAS and Ca9-22 cells and could enhance the ALA-PDT-induced apoptosis of SAS and Ca9-22 cells. We conclude that COX-2 inhibitor combined with ALA-PDT have the synergic effect on the killing of oral cancer cell lines. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T05:28:06Z (GMT). No. of bitstreams: 1 ntu-94-R92422026-1.pdf: 3068410 bytes, checksum: 672e83f7db8094a0755e04f664f204f4 (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 中文摘要 6
ABSTRACT 8 CHAPTER I 10 INTRODUCTION 10 Squamous Cell Carcinoma in Oral Cavity 10 Photodynamic Therapy 12 Photosensitizers 14 5-Aminolevulinic Acid-mediated PDT 16 Oxygen 21 Light Emitting Diodes (LEDs) 21 The Advantages of PDT 22 The Future of PDT 24 Apoptosis 25 PDT-induced Apoptosis 26 Cyclooxygenase-2 ( COX-2 ) 28 COX-2 and Carcinogenesis 29 COX-2 Inhibitor 30 CHAPTER II 33 MATERIALS AND METHODS 33 Drugs and Reagents 33 Cell Lines 33 Photodynamic Therapy 34 Treatment Protocols 34 Cell Viability Assay 35 Preparation of Cellular Protein Extracts 36 Western Immunoblot Analysis 36 Hoechst 33258 Staining 38 DNA Ladder Analysis 38 Statistical Analysis 39 CHAPTER III 40 RESULTS 40 Cyclooxygenase-2 (COX-2) Expression in Oral SCC Cell Lines ………40 COX-2 Expression in Oral SCC Cell Lines after ALA-PDT with/without COX-2 Inhibitor …………………41 Inhibition of Cell Growth by Celebrex ………………………42 Light Does-dependent Inhibition of Growth of SAS and Ca9-22 Cells after ALA-PDT with or without Celebrex …………………………… 43 Induction of Cell Apoptosis by ALA-PDT with or without Celebrex …44 DNA Laddering Analysis ………45 CHAPTER IV 47 DISCUSSION ……………………………………………………………47 CONCLUSIONS ………………………………………54 CHAPTER V (Morphology Assessment) 56 APPENDIX I ( Figures ) ………………………………………57 APPENDIX II ( Abbreviation ) ………………………………………89 REFERENCES ………………………………………………91 | |
dc.language.iso | en | |
dc.title | 光動力合併化學治療對口腔癌細胞株之影響 | zh_TW |
dc.title | Effect of photodynamic therapy plus chemotherapy on oral cancer cell lines | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張龍昌 | |
dc.subject.keyword | 光動力治療,五-氨基酮戊酸,環氧化酵素-2抑制劑,口腔癌,口腔癌細胞株, | zh_TW |
dc.subject.keyword | photodynamic therapy,5-aminolevulinic acid,cyclooxygenase-2 inhibitor,oral cancer,oral cancer cell line., | en |
dc.relation.page | 101 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2005-07-13 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
顯示於系所單位: | 臨床牙醫學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-94-1.pdf 目前未授權公開取用 | 3 MB | Adobe PDF |
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