(3.238.96.184) 您好!臺灣時間:2021/05/12 21:37
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:::

我願授權國圖
: 
twitterline
研究生:李宜靜
研究生(外文):Yi-ching Lee
論文名稱:薑黃素衍生物抑制HER2/neu過度表現癌細胞生長之研究
論文名稱(外文):Curcumin derivatives preferentially inhibit the proliferation of HER2/neu-overexpressing cancer cells
指導教授:魏宗德
學位類別:碩士
校院名稱:中國醫藥大學
系所名稱:生物科技學系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:54
中文關鍵詞:薑黃素表皮生長因子接受器HER2
外文關鍵詞:curcuminEGFRHER2
相關次數:
  • 被引用被引用:0
  • 點閱點閱:275
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
乳癌與卵巢癌是最常被發現的女性惡性腫瘤,發生率佔所有女性癌症的三分之一。HER2基因(又稱為neu或ErbB2)的放大或HER2蛋白的過度表現在乳癌或卵巢癌又佔了將近30%。具有HER2過度表現的乳癌或卵巢癌患者,其治癒率低且復發率高。有研究指出在乳癌或卵巢癌的細胞株若HER2過表現可以促進生長、存活與轉移。薑黃素為一種黃色色素,已有研究指出它具有抗氧化、抗發炎甚至於具有HER2降解的能力。因此,本研究合成了許多薑黃素的衍生物來試驗HER2降解的效果是否會有比薑黃素更好。在本研究中,藉由MTT試驗發現p93cu03與pculin02H具有最佳抑制HER2過表現癌細胞的效果。並且可以誘導細胞週期停滯在G2/M期進一步走向凋亡。本研究也檢測了薑黃素及其衍生物對於HER2蛋白表現量的影響,並發現了衍生物pculin02H對於抑制HER2蛋白的表現量效果優於薑黃素。再者,本研究也想要了解薑黃素衍生物是不是可以促進HER2蛋白的降解,結果顯示薑黃素的衍生物p93cu03可以促進HER2蛋白質降解的速度加快。此外,本研究也利用免疫螢光染色法觀察細胞內HER2蛋白表現量的情況,結果顯示除了控制組以及有轉染HER2基因的細胞其細胞表面仍然有較多的HER2蛋白表現,其餘加了轉譯及轉錄抑制劑跟單純加藥組都有明顯的蛋白質表現下降的現象。接著去探討p93cu03是否會影響細胞轉形(transformation phenotype),藉由固著非依賴性生長檢測,發現 HER2過表現細胞群落形成的狀況很明顯的較HER2未過表現的被抑制了許多。本研究結果可以提供預防或治療HER2過表現癌症的另一有效方法。
Breast carcinoma and ovarian carcinoma are the most frequently diagnosed malignancies, and they account for one-third of all cancers in women. Amplification of the HER2 gene (also known as neu or ErbB2) or overexpression of HER2 protein was found in up to 30% of breast and ovarian carcinomas. Breast and ovarian cancer patients whose tumor cells overexpress HER2 have a poor clinical outcome, such as shorter survival or earlier relapse. HER2 overexpression has been shown to enhance proliferative, prosurvival, and metastatic signals in breast and ovarian cancer cell lines. Curcumin, a yellow pigment, has been reported having the ability of antioxidant, anti-inflammatory, and also having the ability to degrade HER2 protein Therefore, we used the curcumin derivatives to examine whether they have better efficiency to degrade HER2 than curcumin. In the present study, the MTT growth assay showed that p93cu03 and pculin02H had better efficiency to suppress growth of the HER2 -overexpressing cancer cell lines than curcumin. Moreover, p93cu03 and pculin02H preferentially suppressed the growth of HER2-overexpressing cancer cell lines. They both induced G2/M cell cycle arrest followed by apoptosis. In addition, we found that pculin02H has more inhibitory effect in HER2 protein level than curcumin. Interestingly, our results suggested that a posttranslational mechanism contributing to p93cu03 induced HER2 depletion in HER2-overexpressing cancer cells. Furthermore, our results also suggested that p93cu03 significantly more suppressed the colony-forming activity of HER2-overexpressing breast cancer cells than that of nonoverexpressing cell lines. These findings may provide an alternative preventive or therapeutic strategy against HER2-overexpressing cancer cells.
總目錄.............………………………………………………………………..I
圖目錄……………………………………………………………………….IV
符號縮寫表…………………………………………………………………..V
致謝………………………………………………………………………...VII
中文摘要……………………………………………………………………...1
英文摘要……………………………………………………………………...2
第一章 前言.....................................................................................................3
第一節 研究緣起………………………………………………………..3
第二節 文獻回顧………………………………………………………..4
一、 癌症……………………………………………………………4
二、 表皮生長因子接受器家族……………………………………5
三、 癌細胞與HER2的關係………………….................................6
四、 薑黃素…………………………………………………………9
第三節 實驗目的與設計流程………………………………………...13
第二章 研究方法………………………………….………...……………...14
第一節 實驗材料…………………………………………………….14
一、 細胞株………………………………………………………..14
二、 實驗藥物……………………………………………………..14
三、 藥品試劑……………………………………………………..15
四、 設備儀器……………………………………………………..16
第二節 實驗方法………………………………………...…………..18
一、 藥物配製…………………………………………………….18
二、 細胞培養…………………………………………………….18
三、 解凍細胞..................................................................................19
四、 冷凍細胞..................................................................................20
五、 細胞存活率分析 (MTT assay)……………………………...20
六、 流式細胞儀分析 (Flow Cytometry)………………………...21
七、 西方點墨法 (Western blotting)……………………………...22
八、 固著非依賴性生長分析 (Anchorage-Independent Growth Assay).........................................................................................24
九、 免疫螢光染色法(Immunofluorescence Assay)……………...25
十、 細胞轉染(Cell Transfection)…………………………………26
第三章 研究結果…………………………………………………………...27
一、 利用薑黃素衍生物對HER2過表現的卵巢癌細胞進行存活率分析比較…………………………………………………..27
二、 薑黃素衍生物p93cu03與pculin02H對於HER2過表現的卵巢癌細胞週期影響分析……………………………………..27
三、 薑黃素衍生物p93cu03與pculin02H對HER2蛋白質表現量的影響………………………………………………………..28
四、 薑黃素衍生物p93cu03與pculin02H對於不同HER2表現量的細胞株存活率影響分析…………………………………..28
五、 薑黃素衍生物p93cu03促進HER2蛋白降解的研究………29
六、 薑黃素衍生物p93cu03干擾細胞內HER2的表現量的研究……………………………………….…………………….29
七、 薑黃素衍生物p93cu03影響細胞轉形(transformation phenotype)的研究……………………………………………30
第四章 討論………………………………………………………………...31
第五章 結論………………………………………………………………...34
第六章 參考文獻…………………………………………………..……….47
1.行政院衛生署. 十大死因統計; 2009.
2.Hanahan D, Weinberg RA. The hallmarks of cancer. Cell 2000;100(1):57-70.
3.Riese DJ, 2nd, Stern DF. Specificity within the EGF family/ErbB receptor family signaling network. Bioessays 1998;20(1):41-8.
4.Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001;2(2):127-37.
5.Olayioye MA, Neve RM, Lane HA, Hynes NE. The ErbB signaling network: receptor heterodimerization in development and cancer. EMBO J 2000;19(13):3159-67.
6.Schlessinger J. Common and distinct elements in cellular signaling via EGF and FGF receptors. Science 2004;306(5701):1506-7.
7.Muthuswamy SK, Gilman M, Brugge JS. Controlled dimerization of ErbB receptors provides evidence for differential signaling by homo- and heterodimers. Mol Cell Biol 1999;19(10):6845-57.
8.Klapper LN, Waterman H, Sela M, Yarden Y. Tumor-inhibitory antibodies to HER-2/ErbB-2 may act by recruiting c-Cbl and enhancing ubiquitination of HER-2. Cancer Res 2000;60(13):3384-8.
9.Ullrich A, Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell 1990;61(2):203-12.
10.Schechter AL, Hung MC, Vaidyanathan L, et al. The neu gene: an erbB-homologous gene distinct from and unlinked to the gene encoding the EGF receptor. Science 1985;229(4717):976-8.
11.Slamon DJ. Proto-oncogenes and human cancers. N Engl J Med 1987;317(15):955-7.
12.van de Vijver MJ, Mooi WJ, Peterse JL, Nusse R. Amplification and over-expression of the neu oncogene in human breast carcinomas. Eur J Surg Oncol 1988;14(2):111-4.
13.van de Vijver MJ, Mooi WJ, Wisman P, Peterse JL, Nusse R. Immunohistochemical detection of the neu protein in tissue sections of human breast tumors with amplified neu DNA. Oncogene 1988;2(2):175-8.
14.van de Vijver MJ, Peterse JL, Mooi WJ, et al. Neu-protein overexpression in breast cancer. Association with comedo-type ductal carcinoma in situ and limited prognostic value in stage II breast cancer. N Engl J Med 1988;319(19):1239-45.
15.Vaitkevicius P, Wright JG, Fleg JL. Effect of estrogen replacement therapy on the ST-segment response to postural and hyperventilation stimuli. Am J Cardiol 1989;64(16):1076-7.
16.Heintz NH, Leslie KO, Rogers LA, Howard PL. Amplification of the c-erb B-2 oncogene and prognosis of breast adenocarcinoma. Arch Pathol Lab Med 1990;114(2):160-3.
17.Paik S, Hazan R, Fisher ER, et al. Pathologic findings from the National Surgical Adjuvant Breast and Bowel Project: prognostic significance of erbB-2 protein overexpression in primary breast cancer. J Clin Oncol 1990;8(1):103-12.
18.Lodato RF, Maguire HC, Jr., Greene MI, Weiner DB, LiVolsi VA. Immunohistochemical evaluation of c-erbB-2 oncogene expression in ductal carcinoma in situ and atypical ductal hyperplasia of the breast. Mod Pathol 1990;3(4):449-54.
19.Tiwari RK, Borgen PI, Wong GY, Cordon-Cardo C, Osborne MP. HER-2/neu amplification and overexpression in primary human breast cancer is associated with early metastasis. Anticancer Res 1992;12(2):419-25.
20.Wright C, Prasad K, Lennard TJ. erbB2 expression in breast and other human tumours. J Clin Pathol 1992;45(5):459-60.
21.Press MF, Pike MC, Chazin VR, et al. Her-2/neu expression in node-negative breast cancer: direct tissue quantitation by computerized image analysis and association of overexpression with increased risk of recurrent disease. Cancer Res 1993;53(20):4960-70.
22.Hartmann LC, Ingle JN, Wold LE, et al. Prognostic value of c-erbB2 overexpression in axillary lymph node positive breast cancer. Results from a randomized adjuvant treatment protocol. Cancer 1994;74(11):2956-63.
23.Drudis T, Arroyo C, Van Hoeven K, Cordon-Cardo C, Rosen PP. The pathology of low-grade adenosquamous carcinoma of the breast. An immunohistochemical study. Pathol Annu 1994;29 ( Pt 2):181-97.
24.Muss HB, Thor AD, Berry DA, et al. c-erbB-2 expression and response to adjuvant therapy in women with node-positive early breast cancer. N Engl J Med 1994;330(18):1260-6.
25.Giai M, Roagna R, Ponzone R, De Bortoli M, Dati C, Sismondi P. Prognostic and predictive relevance of c-erbB-2 and ras expression in node positive and negative breast cancer. Anticancer Res 1994;14(3B):1441-50.
26.Quenel N, Wafflart J, Bonichon F, et al. The prognostic value of c-erbB2 in primary breast carcinomas: a study on 942 cases. Breast Cancer Res Treat 1995;35(3):283-91.
27.Leal CB, Schmitt FC, Bento MJ, Maia NC, Lopes CS. Ductal carcinoma in situ of the breast. Histologic categorization and its relationship to ploidy and immunohistochemical expression of hormone receptors, p53, and c-erbB-2 protein. Cancer 1995;75(8):2123-31.
28.Leitzel K, Teramoto Y, Konrad K, et al. Elevated serum c-erbB-2 antigen levels and decreased response to hormone therapy of breast cancer. J Clin Oncol 1995;13(5):1129-35.
29.Hieken TJ, Das Gupta TK. Mutant p53 expression: a marker of diminished survival in well-differentiated soft tissue sarcoma. Clin Cancer Res 1996;2(8):1391-5.
30.Xing RH, Rabbani SA. Overexpression of urokinase receptor in breast cancer cells results in increased tumor invasion, growth and metastasis. Int J Cancer 1996;67(3):423-9.
31.Yu D, Liu B, Tan M, Li J, Wang SS, Hung MC. Overexpression of c-erbB-2/neu in breast cancer cells confers increased resistance to Taxol via mdr-1-independent mechanisms. Oncogene 1996;13(6):1359-65.
32.Fernandez Acenero MJ, Farina Gonzalez J, Arangoncillo Ballesteros P. Immunohistochemical expression of p53 and c-erbB-2 in breast carcinoma: relation with epidemiologic factors, histologic features and prognosis. Gen Diagn Pathol 1997;142(5-6):289-96.
33.Moreno A, Lloveras B, Figueras A, et al. Ductal carcinoma in situ of the breast: correlation between histologic classifications and biologic markers. Mod Pathol 1997;10(11):1088-92.
34.Ratcliffe N, Wells W, Wheeler K, Memoli V. The combination of in situ hybridization and immunohistochemical analysis: an evaluation of Her2/neu expression in paraffin-embedded breast carcinomas and adjacent normal-appearing breast epithelium. Mod Pathol 1997;10(12):1247-52.
35.Brien TP, Depowski PL, Sheehan CE, Ross JS, McKenna BJ. Prognostic factors in gastric cancer. Mod Pathol 1998;11(9):870-7.
36.Sjogren S, Inganas M, Lindgren A, Holmberg L, Bergh J. Prognostic and predictive value of c-erbB-2 overexpression in primary breast cancer, alone and in combination with other prognostic markers. J Clin Oncol 1998;16(2):462-9.
37.Jimenez RE, Wallis T, Tabasczka P, Visscher DW. Determination of Her-2/Neu status in breast carcinoma: comparative analysis of immunohistochemistry and fluorescent in situ hybridization. Mod Pathol 2000;13(1):37-45.
38.Seidman A, Hudis C, Pierri MK, et al. Cardiac dysfunction in the trastuzumab clinical trials experience. J Clin Oncol 2002;20(5):1215-21.
39.Bishop JM. The molecular genetics of cancer. Science 1987;235 (4786):305-11.
40.Ross R. Platelet-derived growth factor. Lancet 1989;1(8648):1179-82.
41.Elder JT, Fisher GJ, Lindquist PB, et al. Overexpression of transforming growth factor alpha in psoriatic epidermis. Science 1989;243(4892):811-4.
42.Vassar R, Fuchs E. Transgenic mice provide new insights into the role of TGF-alpha during epidermal development and differentiation. Genes Dev 1991;5(5):714-27.
43.Xu W, Mimnaugh E, Rosser MF, et al. Sensitivity of mature Erbb2 to geldanamycin is conferred by its kinase domain and is mediated by the chaperone protein Hsp90. J Biol Chem 2001;276(5):3702-8.
44.Mimnaugh EG, Chavany C, Neckers L. Polyubiquitination and proteasomal degradation of the p185c-erbB-2 receptor protein-tyrosine kinase induced by geldanamycin. J Biol Chem 1996;271(37):22796-801.
45.Hong RL, Spohn WH, Hung MC. Curcumin inhibits tyrosine kinase activity of p185neu and also depletes p185neu. Clin Cancer Res 1999;5(7):1884-91.
46.張賢哲. 本草備要解析: 中國醫藥學院出版組; 1988.
47.現代本草. 中國藥材學(上.下): 啟業書局; 1976.
48.陳欽銘. 當代中藥學: 協進圖書有限公司; 1982.
49.顏正華. 中藥學(下)高等中醫研究參考叢書14: 知音出版社; 1991.
50.Huang MT, Smart RC, Wong CQ, Conney AH. Inhibitory effect of curcumin, chlorogenic acid, caffeic acid, and ferulic acid on tumor promotion in mouse skin by 12-O-tetradecanoylphorbol-13-acetate. Cancer Res 1988; 48(21):5941-6.
51.Singh S, Aggarwal BB. Activation of transcription factor NF-kappa B is suppressed by curcumin (diferuloylmethane) [corrected]. J Biol Chem 1995;270(42):24995-5000.
52.Surh YJ, Han SS, Keum YS, Seo HJ, Lee SS. Inhibitory effects of curcumin and capsaicin on phorbol ester-induced activation of eukaryotic transcription factors, NF-kappaB and AP-1. Biofactors 2000;12(1-4):107-12.
53.Dorai T, Aggarwal BB. Role of chemopreventive agents in cancer therapy. Cancer Lett 2004;215(2):129-40.
54.Park MJ, Kim EH, Park IC, et al. Curcumin inhibits cell cycle progression of immortalized human umbilical vein endothelial (ECV304) cells by up-regulating cyclin-dependent kinase inhibitor, p21WAF1/CIP1, p27KIP1 and p53. Int J Oncol 2002;21(2):379-83.
55.Jung Y, Xu W, Kim H, Ha N, Neckers L. Curcumin-induced degradation of ErbB2: A role for the E3 ubiquitin ligase CHIP and the Michael reaction acceptor activity of curcumin. Biochim Biophys Acta 2007;1773(3):383-90.
56.Price JE. Clonogenicity and experimental metastatic potential of spontaneous mouse mammary neoplasms. J Natl Cancer Inst 1986;77(2):529-35.
57.Adams BK, Ferstl EM, Davis MC, et al. Synthesis and biological evaluation of novel curcumin analogs as anti-cancer and anti-angiogenesis agents. Bioorg Med Chem 2004;12(14):3871-83.
58.Robinson TP, Ehlers T, Hubbard IR, et al. Design, synthesis, and biological evaluation of angiogenesis inhibitors: aromatic enone and dienone analogues of curcumin. Bioorg Med Chem Lett 2003;13(1):115-7.
59.Youssef KM, El-Sherbeny MA, El-Shafie FS, Farag HA, Al-Deeb OA, Awadalla SA. Synthesis of curcumin analogues as potential antioxidant, cancer chemopreventive agents. Arch Pharm (Weinheim) 2004;337(1):42-54.
60.Jaiswal AS, Marlow BP, Gupta N, Narayan S. Beta-catenin-mediated transactivation and cell-cell adhesion pathways are important in curcumin (diferuylmethane)-induced growth arrest and apoptosis in colon cancer cells. Oncogene 2002;21(55):8414-27.
61.Shim JS, Kim JH, Cho HY, et al. Irreversible inhibition of CD13/aminopeptidase N by the antiangiogenic agent curcumin. Chem Biol 2003;10(8):695-704.
62.Tikhomirov O, Carpenter G. Caspase-dependent cleavage of ErbB-2 by geldanamycin and staurosporin. J Biol Chem 2001;276(36):33675-80.
63.Jana NR, Dikshit P, Goswami A, Nukina N. Inhibition of proteasomal function by curcumin induces apoptosis through mitochondrial pathway. J Biol Chem 2004;279(12):11680-5.
64.d''Azzo A, Bongiovanni A, Nastasi T. E3 ubiquitin ligases as regulators of membrane protein trafficking and degradation. Traffic 2005;6(6):429-41.
65.Liao W, Chan L. Tunicamycin induces ubiquitination and degradation of apolipoprotein B in HepG2 cells. Biochem J 2001;353(Pt 3):493-501.
66.Urbe S. Ubiquitin and endocytic protein sorting. Essays Biochem 2005;41:81-98.
67.Reddy S, Rishi AK, Xu H, Levi E, Sarkar FH, Majumdar AP. Mechanisms of curcumin- and EGF-receptor related protein (ERRP)-dependent growth inhibition of colon cancer cells. Nutr Cancer 2006;55(2):185-94.
68.Pellikainen JM, Ropponen KM, Kataja VV, Kellokoski JK, Eskelinen MJ, Kosma VM. Expression of matrix metalloproteinase (MMP)-2 and MMP-9 in breast cancer with a special reference to activator protein-2, HER2, and prognosis. Clin Cancer Res 2004;10(22):7621-8.
69.Mitsiades N, Yu WH, Poulaki V, Tsokos M, Stamenkovic I. Matrix metalloproteinase-7-mediated cleavage of Fas ligand protects tumor cells from chemotherapeutic drug cytotoxicity. Cancer Res 2001;61(2):577-81.
70.Hynes NE, Stern DF. The biology of erbB-2/neu/HER-2 and its role in cancer. Biochim Biophys Acta 1994;1198(2-3):165-84.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊
 
系統版面圖檔 系統版面圖檔