跳到主要內容

臺灣博碩士論文加值系統

(3.236.84.188) 您好!臺灣時間:2021/08/02 19:05
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:黃俞榮
研究生(外文):Yu-Jung Huang
論文名稱:香椿葉萃取液對非小細胞肺癌細胞株之抗癌功能研究
論文名稱(外文):Anti-cancer Effect of Toona Sinensis on Non-small Cell Lung Carcinoma Cell Lines
指導教授:許勝光黃明賢黃明賢引用關係
指導教授(外文):Hseng-Kuang HsuMing-Shyan Huang
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:醫學研究所碩士班
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:82
中文關鍵詞:非小細胞肺癌保健食品香椿
外文關鍵詞:NSCLCToona sinensishealth food
相關次數:
  • 被引用被引用:12
  • 點閱點閱:354
  • 評分評分:
  • 下載下載:107
  • 收藏至我的研究室書目清單書目收藏:0
在我們過去的研究中,發現香椿葉水萃物(TSL-CE)有效抑制肺腺癌細胞A549的生長。而A549是非小細胞肺癌(NSCLC)中的一個類型,所以我們想了解香椿對人類肺腺癌細胞(H441)、肺鱗狀細胞癌細胞(H520)及肺大細胞癌細胞(H661)等三種非小細胞肺癌(NSCLC)的抗癌影響,結果顯示香椿樹各部份中以香椿葉最有抑癌效果,且NSCLC處理TSL-1(TSL-CE離心後之上清液)後比TSL-CE更有抑制效果,在對肺癌細胞增生之抗癌影響方面。以流式細胞儀分析發現,TSL-1處理三種細胞株的細胞週期都停滯在G1期,進一步以西方墨點法分析調控細胞週期的相關蛋白質分子表現,發現經TSL-1處理後,會致使Cyclin-D1在H441、H520及H661的表現量均下降,cdk4只在H441及H661的表現減少,p27在H520及H661的表現增加,而H661投予TSL-1後,在cylin D1、cdk4及p27表現量變化均較H441和H520明顯,在MTT assay測得IC50之濃度在H661亦是最低,所以TSL-1對NSCLC中的肺大癌細胞(H661)之抑癌效果是最好。在細胞死亡方式之抗癌影響方面,TSL-1處理後的肺癌細胞在細胞型態、Sub-G1的堆積、PARP被裂解情形、以及PI-Annexin-V雙染和TUNEL assay分析之結果,均證實TSL-1會造成三株細胞的凋亡。肺腺癌是NSCLC中最常見的類型,所以在動物試驗,選用H441(肺腺癌細胞)誘發裸鼠皮下腫瘤,分別以治療及預防二種實驗方式來探討,結果發現在治療上不論是灌食或腹腔注射均能有效抑制癌細胞生長;預防實驗中,先期灌食TSL-1亦使腫瘤細胞生長受到抑制而有預防效用。體內試驗之實驗組動物在血液生化值或組織器官上與對照組無差異。此項發現推測口服TSL-1可作為未來研發抗肺癌保健食品之重要學理依據。
由本研究瞭解香椿在NSCLC抗癌能力上,以香椿葉抗癌效果最好,又以其成份TSL-1對於肺大細胞癌細胞(H661)有極佳的抑癌影響及誘發細胞凋亡;長時間餵食TSL-1對體內不會造成副作用,並可治療及預防癌細胞生長。因此TSL-1具有開發成為預防NSCLC之發生及治療用的保健食品。
Our previous study showed that the crude extract from the leaves of Toona sinensis (TSL-CE) exerts potent anti-proliferative effect on A549 lung cancer cells. Since A549 is one of the Non-small-cell lung cancer (NSCLC) types, it is of interest to know whether TS also has anti-cancer effect on all three types of NSCLC. TSL-1, a fraction of TS crude extraction, is the supernatant parts of TSL-CE after centrifugation. The effects of TSL-1 on the proliferation of H441 cells (adenocarcinoma), H661 cells (large cell carcinoma) and H520 cells (squamous cell carcinoma) were investigated in this study. We tested the anti-cancer effect of TSL-1 on the proliferation and cell death. Our results showed that, the anti-proliferative effect of the TSL-1 is more potent than TSL-CE in all three cell types of NSCLC. Treatment of NSCLC cells with TSL-1 resulted in a G1-phase arrest by Flow cytometry. TSL-1 mediated G1 arrest was accompanied by the down-regulation of protein levels of cyclin-D1 in all three cell types of NSCLC, reduced cdk4 expression in H441 and H661, and the up-regulation of p27 in H520 and H661. We found that the decrease in cyclin D1 and cdk4 and the increase in p27 were most significant in H661 cells, compared to other two cell lines. Among the three cell lines, the IC50 for TSL-1 in H661 cells was the lowest, indicating the anti-cancer effect of TSL-1 is most potent in this cell line. TSL-1-induced apoptosis was confirmed by cell morphology, Sub-G1 peak accumulation, cleavage of PARP, PI-Annexin-V double staining and TUNEL assay in all three types of NSCLC. Adenocarcinoma of the lung cancer is now the most worldwide common subtype of NSCLC. Therefore, we performed in vivo experiments to evaluate the therapy and prevention effects of TSL-1 on H441 cells. Per os and intraperitoneal treatment of TSL-1 was used in therapy study, while only per os treatment of TSL-1 was used in prevention study. Our results revealed that treatment of TSL-1 inhibits H441 tumor growth in either therapy or prevention experiments. Whereas, TSL-1 did not show any side effects in these in vivo studies. Our results suggest that TSL-1 could be used as a health food for preventing and alleviating lung cancer proliferation and possess the potential to be developed as an anticancer drug.
中文摘要……………………...…………................................................I
英文摘要…………...…………………………………………………...Ⅲ
縮寫表 (Abbreviations)…………………………………………….….Ⅴ

第一章:緒論 (Introduction)
1.1 香椿 (Toona sinensis)…………………………………....1
1.2 肺癌 (Lung cancer)………………………………………2
1.3 肺癌之細胞週期與細胞凋亡(Cell cycle and Apoptosis)..4
1.4 研究目的 (Purpose)…………………………………......10

第二章:材料與方法 (Materials and methods)……………………….11
第一部分:探討香椿成分TSL-1對非小細胞肺癌細胞株
的抑癌影響…………………………………..…13
第二部分:探討香椿成分TSL-1在活體動物抗癌及防癌
作用……………………………………………..25

第三章:結果 (Results)………………………………………………..31
第一部分:探討香椿成分TSL-1對非小細胞肺癌細胞株
的抑癌影響……………………………………..31
第二部分:探討香椿成分TSL-1在活體動物抗癌及防癌
作用……………………………………………..37

第四章:討論 (Discussion)……………………….…………………...39

第五章:參考文獻 (References)………………………………………46

第六章:圖表…………………………………………………………...53


附錄 學會論文發表………………………………………………....76
中文參考文獻

Chang HC, Yeh CH, Huang MC, Hsu C, and Hsu HK: 香椿萃取液減少Bcl 2蛋白質表現而促進H520肺癌細胞自然凋亡。Health food society of Taiwan. 2001; Feb.30.

Chen WI, Chen YR, Wang PH, Lue SI and Hsu HK: 椿葉萃取物對血小板凝集的作用。Health food society of Taiwan. 2002; Feb.70.

Tsai LY, Huang CC, Hsiao JK, Lin SK, and Chang CS: Effects of Toona sinensis Roemor Concentrate on H2O2-induce Oxidative Damage in HL-60 and K562 Cell Lines. 中華民國醫事檢驗學會九十二年度學術發表會,2003/12/13-14,台北榮總致德樓.

張慧秋:探討銀合歡種子內含成分mimosine在試管內與活體動物抗癌作用之分子機制。高雄醫學大學醫學研究所博士論文。(2000)。

王珮憓:椿葉水萃取液在Alloxan所誘發糖尿病鼠中降血糖作用之研究。高雄醫學大學醫學研究所碩士論文。(2001)。

楊馥瑜:香椿對人類精子生理功能之探討。成功大學生物學系研究所碩士論文。(2003)。

徐雯敏:香椿對老化促進小鼠學習記憶能力、抗氧化狀態與精子品質之影響。靜宜大學食品營養學系碩士論文。(2004)。

徐沛甄,許勝光,王銘富,廖俊旺,詹吟菁:香椿葉萃取物對老化促進小鼠腦部氧化狀態及老化病理變化之影響。台灣農化與食品科學,2003。

徐沛甄,徐雯敏,許勝光,廖俊旺,王銘富,詹吟菁:香椿葉萃取物對老化促進小白鼠學習記憶能力之影響。中華民國營養學會第29屆年會暨學術研討會手冊,2003。

楊今詳:抗癌中草藥製劑。渡假出版社有限公司,台北台灣;1992;165頁。
程劍華、李以鑌:抗癌植物藥及其驗方。江西科學出版社,中國;1998;608頁。


英文參考文獻

Bartkova J, Lukas J and Bartek J: Aberrations of the G1-and G1/S-regulating genes in human cancer. Prog. Cell Cycle Res. 1997; 3, 211-220.

Bishop JM: Molecular themes in oncogenesis. Cell. 1991; 64, 235-240.

Blain SW, Scher HI, Cordon-Cardo C, and Koff A: p27 as a target for cancer therapeutics. Cancer Cell. 2003; Feb, 3(2), 111-5.

Bodver W, Bishop T, and Karran P: Genetic steps in colorectal cancer. Nat Genet 1994; 6, 217-222.

Cantley LC, Auger KR, Carpenter C, Duckworth B, Graziani A, Kapeller R, and Soltoff S: Oncogenes and signal transduction. Cell 1991; 64, 281-285.

Catzavelos C, Taso MS, DeBoer G, Bhattacharya N, Shepherd FA and Slingerland JM: Reduced expression of the cell cycle inhibitor p27Kip1 in non-small cell lung carcinoma: a prognostic factor independent of Ras. Cancer Res. 1999; 59, 684-688.

Chang HC, Weng CF, Yen MH, Chuang LY, Hung WC: Modulation of cell cycle regulatory protein expression and suppression of tumor growth by mimosine in nude mice. Int J Oncol. 2000: Oct, 17 (4), 659-65.

Chang HC, Hung WC, Huang MS and Hsu HK:Extract from the leaves of Toona sinensis exerts potent anti-proliferative effect on human lung cancer cells. Am J Chinese 2002; Med, 30, 2&3, 307-314.

Chang HC, Huang YC, and Hung WC: Antiproliferative and chemopreventive effects of adlay seed on lung cancer in vitro and in vivo. J Agric Food Chem. 2003; Jun 4; 51 (12), 3656-60.

Chang HC, Chang FR, Wu YC, and Lai YH: Anti-cancer effect of liriodenine on human lung cancer cells. Kaohsiung J Med Sci. 2004; Aug, 20 (8), 365-71.

Cao MY, Lee Y, Feng NP, Al-Qawasmeh RA, Viau S, Gu XP, Lau L, Jin H, Wang M, Vassilakos A, Wright JA, and Young AH: NC381, a novel anticancer agent, arrests the cell cycle in G0-G1 and inhibits lung tumor cell growth in vitro and in vivo. J Pharmacol Exp Ther. 2004; Feb, 308 (2), 538-46.

Chen PL, Scully P, Shew JY, Wang J, and Lee WH: Phosphorylation of the retinoblastoma gene product is modulated during the cell cycle and cellular differentiation. Cell 1989; 58, 1193-1199.

Chiba I, Takahashi T, Nau MM, D’Amico D, Curiel DT, Mitsudomi T, Buchhagen DL, Carbone D, Piantadosi S, and Koga H: Mutaions in the p53 gene are frequent in primary, resected non-small cell lung cancer. Oncogene 1990; 5, 1603-1610.

Chiu NY, Technician, and Chang KH:Toona sinensis(A.Juss) Roem. In the ullustrated medicinal plants of Taiwan, SMC publishing Inc Taiwan ROC 1995: 1st, 131-132.

D’Amico D, Carbone D, and Mitsudomi T: High frequency of somatically acquired p53 mutations in small cell lung cancer cell lines and tumors. Oncogene 1992; 7, 339-346.

Doronin K, Kuppuswamy M, Toth K, Tollefson AE, Krajcsi P, Krougliak V, Wold WS: Tissue-specific, tumor-selective, replication-competent adenovirus vector for cancer gene therapy. J Virol. 2001; Apr, 75(7), 3314-24.

Edmonds JM, and Staniforth M:Toona Sinensis(Meliaceae). Curtis’s Botanical Magazine 1998; 15 (3), 186-193.

Elledge RM, Fuqua SA, Clark GM, Pujol P, Allred DC, and McGuire WL: Prognostic significance of p53 gene alterations in node-negative breast cancer. Breast Cancer Res Treat 1993; 26, 225-229.

Fan S, el-Deiry WS, Bae I, Freeman J, Jondle D, Bhatia K, Fornace AJ Jr, Magrath I, Kohn KW, and O''Connor PM:p53 gene mutations are associated with decreased sensitivity of human lymphoma cells to DNA damaging agents. Cancer Res. 1994; Nov 15, 54(22), 5824-30.

Haber DA. Splicing into senescence: the curious case of p16 and p19ARF. Cell 1997; 91, 555–558.

Hsieh TC and Wu JM: Apoptosis and restriction of G1/S cell cycle by Fenretide in Burkitt’s lymphoma Mutu 1 cell line accessed with bcl-6 down-regulation. Biochemical and Biophysical Research Communication 2000; 276, 1295-1301.

Hsieh TJ, Liu TZ, Chia YC, Chern CL, Lu FJ, Chuang MC, Mau SY, Chen SH, Syu YH, and Chen CH: Protective effect of methyl gallate from Toona sinensis (Meliaceae) against hydrogen peroxide-induced oxidative stress and DNA damage in MDCK cells. Food Chem Toxicol 2004; May, 42 (5), 843-50.

Ikegami Y, Yano S, and Nakao K: Antitumor effect of CGP41251, a new selective protein kinase C inhibitor, on human non-small cell lung cancer cells. Jpn J Pharmacol. 1996; Jan, 70 (1), 65-72.

Jennifer M, Edmonds, Martin S: Toona Sinensis. Bentham-Moxon Trust. 1998; 186-196.

Jiang W, Kahn SM, Tomia N, Zhang YJ, Lu SH, and Weinstein IB: Amplification and expression of the human cyclin D gene is esophageal cancer. Cancer Res 1992; 52, 2980-2986.

John Kerr and apoptosis. Med. J. 2000; Aust. 173 (11-12), 616-617.

Kamesaki H: Mechanisms involved in chemotherapy-induced apoptosis and their implications in cancer chemotherapy, Int. J. Hematol. 1998; 68, 29-43.

Kerr JF, Winterford CM, and Harmon BV, Apoptois. Its significance in cancer and cancer therapy, Cancer 1994; 73 (8), 2013-2026.

Khan QA, and Anderson LM: Natural protection from apoptosis by surfactant protein A in type Ⅱ pneumocytes. Exp Cell Res. 2001; Feb 15, 263 (2), 183-192.

Labuda J, Buckova M, Heilerova L, Silhar S, Stepanek I: Evaluation of the redox properties and anti/pro-oxidant effects of selected flavonoids by means of a DNA-based electrochemical biosensor. Anal Bioanal Chem. 2003; 376, 168-173.

Lai RS, Wang JS, Hsu HK, Chang HC, Lin CH and Lin MH:Prognstic evaluation of the expression of p53 and bcl-2 oncoprotein in patients with surgically resected non-small cell lung cancer. Jpn J. Clin. Oncol. 2002; 32 (10), 393-397.

Lee JH, Lee CT, Yoo CG, Hong YK, Kim CM, Han SK, Shim YS, Carbone DP, and Kim YW: The inhibitory effect of adenovirus-mediated p16INK4a gene transfer on the proliferation of lung cancer cell line. Anticancer Res. 1998; Sep-Oct, 18 (5A), 3257-61.

Liu LF, Liang CH, Shiu LY, Lin WL, Lin CC, Kuo KW: Action of solamargine on human lung cancer cells--enhancement of the susceptibility of cancer cells to TNFs. FEBS Lett. 2004; Nov 5, 577(1-2), 67-74.

Merlo A, Hermann JG, and Mao L: 5*CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers. Nature Med 1995; 1, 686–692.

Moody TW, and Jensen RT: CI-988 Inhibits Growth of Small Cell Lung Cancer Cells. J Pharm Exp Ther. 2001; 299, 1154–1160.

Narang D, Sood S, Thomas MK, Dinda AK, Maulik SK: Effect of dietary palm olein oil on oxidative stress associated with ischemic-reperfusion injury in isolated rat heart. BMC Pharmacol. 2004; Nov 09, 4 (1), 29.

O''Connor PM:Mammalian G1 and G2 phase checkpoints, Cancer Surv. 1997; 29, 151-182.

Park JC, Yu YB Yu, Lee JH, Chol JS, and Ok KD:Phenolic Compounds form the Rachis of Cedrela sinensis. Kor J Pharmacogn, 1996; 27 (3), 219-223.

Reed SI, Bailly E, Dulic V, Hengst L, Resnitzk D, and Slingerland J:G1 control in mammalian cells, J. Cell Sci. Suppi. 1994; 18, 69-73.

Reed JC:Apoptosis-regulating proteins as targets for drug discovery, Trends Mol. 2001; Med, 7, 314-319.

Rosenberg Cl, Wong E, Petty WM, Bale AE, Tsujimoto Y, Harris NL, andArnold A: PRAD1, a candidate BCL1 oncogene: Mapping and expression in centrocytic lymphoma. Proc Natl Acad Sci 1991; 88, 9638-9643.

Scudiero DA, Shoemaker RH, Paull KD, Monks A, Tierney S, Nofziger TH, Currens MJ, Seniff D, Boyd MR. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res. 1988; Sep 1, 48(17), 4827-33.

Shafer SH, and Williams CL: Non-small and small cell lung carcinoma cell lines exhibit cell type-specific sensitivity to edelfosine-induced cell death and different cell line-specific responses to edelfosine treatment. Int J Oncol. 2003; Aug, 23 (2), 389-400.

Shapiro GI, Edwards CD, Kobzik L, Godleski J, Richards W, Sugarbaker DJ, and Rollins BJ: Reciprocal Rb inactivation and p16INK4 expression in primary lung cancers and cell lines. Cancer Res. 1995; 55, 505-509.

Shapiro GI, Park JE, Edwards CD, Mao L, Merlo A, Sidransky D, Ewen ME, Rollins BJ: Multiple mechanisms of p16INK4A inactivation in non-small cell lung cancer cell lines. Cancer Res. 1995; Dec 15, 55 (24), 6200-6209.

Sherr CJ. Cancer cell cycle. Science 1996; 274, 1672-1677.

Singh N, Sun Y, Nakamura K, Smith MR, and Colburn NH: C-JUN/AP-1 as possible mediators of tumor necrosis factor-alpha-induced apoptotic response in mouse JB6 tumor cells. Oncol Res 1995; 7 (7-8), 353-362.

Stuschke M, Sak A, Wurm R, Sinn B, Wolf G, Stuben G, and Budach V: Radiation-induced apoptosis in human non-small-cell lung cancer cell lines is secondary to cell-cycle progression beyond the G2-phase checkpoint. Int J Radiat Biol. 2002; Sep, 78 (9), 807-19.

Takano YS, Harmon BV, and Kerr JF:Apoptosis induced by mild hyperthermia in human and murine tumour cell lines: a study using electron microscopy and DNA gel electrophoresis. J Pathol. 1991; Apr, 163 (4), 329-336.

Weinberg RA. The retinoblastoma protein and cell cycle control. Cell 1995; 81, 323-330.

Wyllie AH, Kerr JF, and Currie AR: Cell death:the significance of apoptosis. Int Rev Cytol. 1980; 68, 251-306.

Xu XJ, Hokfelt T, Hughes J, and Wiesenfeld-Hallin Z: The CCK-B antagonist CI988 enhances the reflex-depressive effect of morphine in axotomized rats. Neuroreport. 1994; Feb 24, (6), 718-20.

Yatabe Y, Masuda A, Koshikawa T, Nakamura S, Kuroishi T, Osada H, Taahashi T, Mitsudomi T, and Takahashi T: p27KIP1 in human lung cancers: differential changes in small cell and non-small cell carcinomas. Cancer Res 1998; 58, 1042-1047.

Yuan J, Knorr J, Altmannsberger M, Goeckenjan G, Ahr A, Scharl A, and Strebhardt K. Expression of p16 and lack of pRB in primary small cell lung cancer.J Pathol. 1999; Nov, 189 (3), 358-62.

Zhang YJ, Jiang W, Chen CJ, Lee CS, Kahn SM, Santella RM, and Weinstein IB:Amplification and overexpression of cyclin D1 in human hepatocellular carcinoma. Biochem Biophys Res Commun 1993; 196 (2), 1010-1016.

Zhang YA, Nemunaitis J, Scanlon KJ, Tong AW: Anti-tumorigenic effect of a K-ras ribozyme against human lung cancer cell line heterotransplants in nude mice. Gene Ther. 2000; Dec, 7(23), 2041-50.

Zochbauer-Muller S, Gazdar, A F, and Minna J D: Molecular pathogenesis of lung cance. Ann. Rev Physiol. 2002; 64, 681-708.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top