Responsive image
博碩士論文 etd-0814112-153336 詳細資訊
Title page for etd-0814112-153336
論文名稱
Title
天然化合物AN30於口腔癌細胞上的毒性與蛋白質體學分析
Cytotoxicity and Proteomes Studies of AN30 on Oral Cancer Cell
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
32
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2012-07-25
繳交日期
Date of Submission
2012-08-14
關鍵字
Keywords
蛋白質體學、口腔癌、AN30、天然化合物
oral cancer, proteomes, natural compound
統計
Statistics
本論文已被瀏覽 5692 次,被下載 6
The thesis/dissertation has been browsed 5692 times, has been downloaded 6 times.
中文摘要
天然化合物代號AN30從蕨類植物(Thelypteris torresiana)純化,在本實驗中證實對口腔癌細胞(Ca9-22)及正常口腔細胞(HGF-1)有效。根據文獻指出AN30在乳腺癌、前列腺癌及肺癌都能有效的誘導細胞凋亡現象並且使細胞週期停滯的現象造成DNA的破壞,但這些現象都出現於腺體癌症上並未有實驗出現在非腺體癌症上應用。所以此實驗藉由口腔癌細胞Ca9-22(鱗狀細胞癌)來進行實驗並了解其作用機制。實驗中已初步證AN30對口腔癌細胞在48小時呈現毒性但對正常的口腔細胞則無。並利用comet assay 及western blot證實細胞凋亡現象,利用流式儀觀察細胞週期有S-phase停滯的現象。且於二維電泳中分析發現約有20個差異點將更進一步經由LC/MS來鑑定其蛋白質身分。
Abstract
The effect of natural compound AN30 on oral cancer cell line (Ca9-22), and normal oral cell line (HGF-1) was investigated. AN30 was purified from native fern plant Thelypteris torresiana. It had been reported to induce cell cycle arrest, apoptosis and DNA damage, cell growth inhibition in lung cancer cells, breast cancer cells and prostate cancer cells, but the effect in oral cancer is unknown. In a preliminary test, AN30 shows high toxicity to oral cancer cells but not normal oral cells. After treatment of AN30 to oral cancer cells, caspase activation and comet assays were performed to verity apoptosis induction. 2-D electrophoresis was used to find out the differentially expressed proteins between normal and cancer oral cell line under AN30 treatment. The proteins were further identified by LC/MS and western blot. The mechanism of these candidate proteins on drug toxicity remains further investigation.
目次 Table of Contents
英文縮寫表 I
中文摘要 II
英文摘要 III
壹、序言
1、口腔癌介紹…………………………………………………………………….10
(1)口腔癌誘因及病理
(2)口腔癌治療方式
2、天然萃取物於癌症上使用................................................................................12
(1)天然萃取物的過去研究
(2)AN30的介紹及結構式
3、蛋白質體學的優勢……………………………………………………………13
貳、實驗流程 (Experiment flowchart) ……………………………......................14
参、材料與方法 (Materials and Methods) …………………………………........15
1.細胞條件與藥物配置
(1)天然化合藥物(AN30)配置
(2)細胞株
(3)細胞培養
2.二維電泳
(1)細胞蛋白質萃取
(2)一維電泳
(3)膠條平衡
(4)藍染及銀染
3.西方點墨法
(1)蛋白質萃取與定量
(2)蛋白質電泳
4. MTT assay
5.Comet assay 實驗
6.流式細胞儀分析
肆、實驗結果……………………………………………………………………20
1. MTT assay 測出IC50與對正常細胞(HGF-1)細胞毒性
2. AN30誘導癌症細胞凋亡並使DNA產生斷裂現象
3. AN30所誘導Caspase活化機制
4. AN30誘導細胞週期停滯於S-phase
伍、實驗討論……………………………………………………………………22
1. AN30細胞毒性分析
2.彗星實驗(Comet assay)對DNA斷裂的探討
3. AN30會使口腔癌細胞造成細胞凋亡現象
4.流式細胞儀顯現出sub-G1的增加及細胞周期停滯於S-phase
陸、圖表…………………………………………………………………………24
Figure 1. MTT assay of AN30 on Ca9-22 and HGF-1 cell line
Figure 2. Western blotting of PARP and caspase3 of treated Ca9-22 cell
Figure3 . Comet assay of AN30 treated Ca9-22 cells .
Figure 4. AN30 induce sub-G1 accumlation and cell cycle arrest in Ca9-22
Figure 5. 2-D Electrophoresis Analysis.
柒、參考文獻……………………………………………………………………30
參考文獻 References
1.Epidemiology and prevention of head and neck cancer: a global view. Contemporary issues in oral cancer. 2000
2.Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery. Mol. Cancer. 2006;5:20
3.The signaling pathway of taxol-induced apoptosis in human oral cancer cell line. 國立成功大學 藍宇彥 碩士論文
4.Jia LQ,et al. Screening the p53 status of human cell lines using a yeast functional assay. Mol. Carcinog. 1997;19:243-53.
5.Sang-Heng Koket al.Norcantharidin-induced apoptosis in oral cancer cells is associated with an increase of proapoptotic to antiapoptotic protein ratio. Cancer Lett. 2005 ;217 :43-52
6.Fang-Rong Chang,et al.Anti-inflammatory and cytotoxic diterpenes from Formosan Polyalthial longifolia var.pendula. Planta Med. 2006 ;72: 1344-7
7.Klaus Schulze-Osthoff,et al.New approaches and therapeutics targetin apoptosis in disease [review]. Pharmacol Rev. 2005 ; 57 :187-215
8.Jun-Hee Kim,et al.Apoptotic EffectlofeTolfenamic Acid in KB Human Oral Cancer Cells: Possible Involvement of the p38 MAPK Pathway. Clin. Biochem. Nutr. 2010 ; 47: 74–80.
9.Ching-Yu Yen,et al.4β-Hydroxywithanolide E from Physalis peruviana (golden berry) inhibits growth of human lung cancer cells through DNA damage, apoptosis and G2/M arrest. BMC Cancer. 2010;10:46.
10. Funk GF,et al. Presentation, treatment, and outcome of oral cavity cancer: a National Cancer Data Base report. Head and Neck. 2002 ;24:165-80
11.The apoptoic mechanism of 4β-hydroxywithanolide E in human A549 cells.
嘉南藥理科技大學 鐘珮蓉 碩士論文
12. Ying-Ray Lee,et al.Reversine suppresses oral squamous cellcarcinoma via cell cycle arrest and concomitantly apoptosis and autophagy. Journal of Biomedical Science. 2012;19:9
13. Cheng-Chang Yeh,et al.DR5 expression cancer cells to TRAIL-induced apoptosis through increaseSuberoylanilide hydroxamic acid sensitizes human oral. Mol. Cancer Ther. 2009 ; 8: 2718.
14. Ye H,et al.Proteomic Based Identification of Manganese Superoxide Dismutase 2 (SOD2) as a Metastasis Marker for Oral Squamous Cell Carcinoma.CANCER GENOMICS& PROTEOMICS .2008 ;5:85-94.
15.Noor Hasima Nagoor,et al.Regulation of Apoptotic Effects by Erythrocarpine E, a Cytotoxic Limonoid from Chisocheton erythrocarpus in HSC-4 Human Oral Cancer Cells. PLoS ONE. 2011; 6: e23661.
16. Tsai WC,et al.Methylantcinate A induces tumor specific growth inhibition in oral cancer cells via Bax-mediated mitochondrial apoptotic pathway. Bioorganic & Medicinal Chemistry Letters. 2010 ;20:6145-8.
17. Chen YW,et al.Triptolide exerts anti-tumor effect on oral cancer and KB cells in vitro and in vivo. Oral Oncology . 2009 ;45:562-8
18. Young-Man Lee,et al.Isoliquiritigenin 20-methyl ether induces growth inhibition and apoptosis in oral cancer cells via heme oxygenase-1. Toxicology in Vitro. 2010; 24:776-82.
19. Chien-Chih Chiu,et al.Fern Plant–Derived Protoapigenone Leads to DNA Damage, Apoptosis, and G2/M Arrest in Lung Cancer Cell Line H1299. DNA and CELL BIOLOGY .2009; 28: 501-506.
20. Chang HL,et al.Protoapigenone, a Novel Flavonoid, Induces Apoptosis in Human Prostate cancer Cells through Activation of p38 Mitogen-Activated Protein Kinase and c-Jun NH2-Terminal Kinase 1/2.Journal of Pharmacology and Experimental Therapeutics . 2008 ;325:841-9.
21. Chen WY,et al.Protoapigenone, a natural derivative of apigenin, induces mitogen-activated protein kinase-dependent apoptosis in human breast cancer cells associated with induction of oxidative stress and inhibition of glutathione S-transferase π. Invest. New Drugs. 2011 ;29:1347-59.
22. Yang-Hao Yu,et al.Ganoderma tsugae Induces S Phase Arrest and Apoptosis in Doxorubicin-Resistant Lung Adenocarcinoma H23/0.3 Cells via Modulation of the PI3K/Akt Signaling Pathway. Evidence-Based Complementary and Alternative Medicine. 2012; 37:1286.
23. Yu,C.C,et al. Norcantharidin triggers cell death and DNA damage through S-phase arrest and ROS-modulated apoptotic pathways in TSGH 8301 human urinary bladder carcinoma cells. Int. J. Oncol. 2012; 41:1050-60.
電子全文 Fulltext
本電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。
論文使用權限 Thesis access permission:自定論文開放時間 user define
開放時間 Available:
校內 Campus: 已公開 available
校外 Off-campus:永不公開 not available

您的 IP(校外) 位址是 54.226.126.38
論文開放下載的時間是 校外不公開

Your IP address is 54.226.126.38
This thesis will be available to you on Indicate off-campus access is not available.

紙本論文 Printed copies
紙本論文的公開資訊在102學年度以後相對較為完整。如果需要查詢101學年度以前的紙本論文公開資訊,請聯繫圖資處紙本論文服務櫃台。如有不便之處敬請見諒。
開放時間 available 已公開 available

QR Code