跳到主要內容

臺灣博碩士論文加值系統

(3.236.50.201) 您好!臺灣時間:2021/08/05 20:52
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:王怡勤
研究生(外文):Wang,Yi-Chin
論文名稱:探討紫米粗萃花青素抑制人類口腔舌癌CAL 27細胞之轉移機制
論文名稱(外文):Study of the mechanisms of anthocyanidins from crude black rice extract in human oral tongue cancer CAL 27 cells
指導教授:范宗宸鍾景光鍾景光引用關係
指導教授(外文):Fan,Ming-JenChung,Jing-Gung
口試委員:范宗宸鍾景光楊家欣
口試委員(外文):Fan,Ming-JenChung,Jing-GungYang,Jai-Sing
口試日期:2012-06-28
學位類別:碩士
校院名稱:亞洲大學
系所名稱:生物科技學系碩士班
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:2012
畢業學年度:100
語文別:中文
論文頁數:88
中文關鍵詞:花青素口腔癌轉移
外文關鍵詞:anthocyanidinsoral cancermetastasis
相關次數:
  • 被引用被引用:3
  • 點閱點閱:460
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:2
有色米的色素主要分佈於穎果皮或是種皮,其色素主要是由花青素所構成。植物主要的花青素有紫紅色的矢車菊素、橘紅色的天竺葵素及藍紫色的飛燕草素等三種,均為抗氧化物,可抑制脂質過氧化,並具有清除自由基的能力。HPLC分析結果顯示,紫米種皮粗萃物主要的花青素是cyanidin-3-glucoside。癌細胞的侵襲及轉移往往是癌症病患致死且導致預後不良的主要原因之一,而其侵入則與細胞間質的分解、細胞的貼附能力及移動性的改變有關。花青素是一種天然的多酚類,廣泛的存在於日常食用的蔬果中,有許多文獻指出花青素在癌症的化學預防上已有許多顯著的生物活性,具有促使腫瘤細胞凋亡及預防正常細胞癌化等作用。因此,對癌症治療有很大的研究價值。腫瘤轉移是癌症死亡最主要的原因,很多治療方式都是針對防止轉移的發生。使用trans well migration/invasion assay檢測CAL 27細胞侵襲/轉移能力,發現紫米粗萃花青素可以抑制CAL 27細胞遷移和侵襲的能力。另外明膠蛋白酵素電泳顯示,花青素能抑制基質金屬蛋白酶2(matrix metalloproteinase 2)的活性。西方墨點法分析顯示花青素抑制CAL 27細胞轉移/侵襲相關蛋白的表現。另外,免疫螢光染色證實,花青素能抑制細胞核轉錄因子p65(NF-κB p65)的表現。基於上述結果,我們證實紫米粗萃花青素藉由抑制PI3K/Akt和NF-κB路徑可以減少MMP-2、MMP-9、u-PA與NF-κB p65表現而去抑制CAL 27細胞轉移。
There are many colored rice in rice germplasm. The pigments of colored rice are mainly located at the pericarp or seed coats of rice grain and they are anthocyanins pigments. The red cyanidins, orange pelargonidin, and purple-blue delphinidin are three major anthocyanins, which are reported to have anti-oxidative activity on lipid and scavenge activity for removing free radical. The results of HPLC analyses showed that the major anthocyanin is cyanidin-3-glucoside in the pericarp of black rice. Metastasis, the major cause of cancer death and various treatment strategies have targeted on preventing the occurrence of metastasis. The invasion of metastasis is connected with proteolytic degradation of the extracellular matrix(ECM) and cell adhesion, essential to achieving cell motility. Anthocyanins, a large group of natrual polyphenols existing in a wide range of daily fruits and vegetables, have been widely recognized to possess several potential as cancer chemopreventive agents. Recent studies have revealed pleiotropic anticancer and antiproliferative capabilities of anthocyanins. Therefore, it may be of great value in developing a potential cancer therapy for anthocyanins. Tumor metastasis is the most important cause of cancer death and various treatment strategies have targeted on preventing the occurrence of metastasis. The anti-metastatic mechanism of anthocyanins on the invasion/migration of human oral CAL 27 cells which were performed by used a Transwell to quantify the migratory potential of CAL 27 cells and we found that anthocyanins can inhibit in vitro migration and invasion of CAL 27 cancer cells. Besides, the gelatin zymography assay indicated that anthocyanins inhibited the activity of matrix metalloproteinases 2(MMP-2). Western blotting assay also showed that anthocyanins inhibited the associated protein expression of migration/invasion of CAL 27 cell. Immunofluorescence staining proved that anthocyanins inhibited Nuclear factor kappa B-p65(NF-κB p65)expressions. These results demonstrated that anthocyanins could suppress CAL 27 cell metastasis by reduction of MMP-2, MMP-9,u-PA and NF-κB p65 expression through the suppression of PI3K/Akt pathway and inhibition of NF-κB levels.
總目錄 Ⅰ
表目錄 Ⅳ
圖目錄 Ⅴ
縮寫表 Ⅶ
第一章 前 言
第一節 研究背景 1
一、口腔癌 1
二、紫米 5
三、花青素 6
四、細胞轉移與侵襲 9
五、基質金屬蛋白酶 11
六、細胞核轉錄因子-κB 12
第二節 研究目的 14
第二章 材料與方法
第一節 實驗材料 15
一、細胞來源 15
二、藥品試劑 15
三、儀器設備 18
第二節 實驗方法 20
一、花青素萃取 20
二、紫米花青素分析 21
三、藥品配製 22
四、人類口腔舌癌細胞株CAL 27培養 22
五、細胞存活率分析 24
六、細胞生長抑制分析 26
七、細胞傷口癒合試驗 27
八、細胞轉移試驗 27
九、細胞侵襲試驗 28
十、細胞貼附試驗 29
十一、明膠酵素電泳法 29
十二、西方墨點法 32
十三、免疫螢光染色 37
十四、電泳位移測定 38
十五、統計分析 39
第三章 研究結果
第一節 紫米花青素高效液相層析分析圖譜 40
第二節 紫米花青素對人類口腔舌癌細胞(CAL 27)存活率的影響 41
第三節 紫米花青素對人類口腔舌癌細胞(CAL 27)增生的影響 43
第四節 紫米花青素對人類口腔舌癌細胞(CAL 27)移行的影響 45
第五節 紫米花青素對人類口腔舌癌細胞(CAL 27)貼附能力的影響 47
第六節 紫米花青素對人類口腔舌癌細胞(CAL 27)轉移能力的影響 49
第七節 紫米花青素對人類口腔舌癌細胞(CAL 27)細胞侵襲能力的影響 51
第八節 紫米花青素對人類口腔舌癌細胞(CAL 27)之MMPs活性的影響 53
第九節 紫米花青素對人類口腔舌癌細胞(CAL 27)之之轉移相關蛋白的影響 54
第十節 利用EMSA探討紫米花青素對CAL 27細胞NF-κB與核內DNA結合變化 60
第十一節 利用免疫螢光染色探討紫米花青素對人CAL 27細胞NF-κB蛋白的表現與轉位之影響 61
第四章 討 論 62
第五章 結 論 66
第六章 參考文獻 68
1.行政院衛生署【cite;http://www.doh.gov.tw/.(2011)】
2.世界衛生組織【cite;http://www.who.int/en/.(2011)】
3.台中榮民總醫院【cite;http://www.vghtc.gov.tw/home/index.html】
4.國家衛生研究院【cite;http://www.nhri.org.tw/】
5.口腔癌分期圖【cite;http://dentallecnotes.blogspot.com】
6.American Joint Committee on Cancer,AJCC。【cite;http://www.cancerstaging.org/】
7.紫米【cite;http://www.asia.edu.tw/main_pages/hot_news/news_09_11_18_1.htm】
8.李勣等修。新修本草(唐)。昭和43[民57]
9.湖北省中醫藥研究院醫史文獻研究室,本草鋼目精要(1988),廣東科技出版發行。
10.(元)李杲編輯/(明)李時珍參訂/(明)姚可成補輯/鄭金生,食物本草(1990),中國醫藥科技出版發行。
11.孔伯華養生醫館,食療本草經草(2006),中國樺齡出版社。
12.達美君、樓紹來,食物本草點校本(1994),人民衛生出版社。
13.Kong, S., D. J. Kim, et al. (2012). "Black rice bran as an ingredient in noodles: chemical and functional evaluation." J Food Sci 77(3): C303-307.
14.Madani, A. H., M. Dikshit, et al. (2012). "Risk for oral cancer associated to smoking, smokeless and oral dip products." Indian J Public Health 56(1): 57-60.
15.Javed, F., K. Al-Hezaimi, et al. (2012). "Areca-nut chewing habit is a significant risk factor for metabolic syndrome: a systematic review." J Nutr Health Aging 16(5): 445-448.
16.Tempesta, M. S. (2012). "Review of Plant Phenolics and Human Health: Biochemistry, Nutrition and Pharmacology Plant Phenolics and Human Health: Biochemistry, Nutrition and Pharmacology . Edited by C. G. Fraga (Universities of Buenos Aires and California-Davis). John Wiley and Sons : Hoboken, NJ, USA . 2010 . xii + 593. 16 x 24 cm. $135.00. ISBN 978-0-470-28721-7." J Nat Prod 75(6): 1260.
17.Weng, C. J. and G. C. Yen (2012). "Flavonoids, a ubiquitous dietary phenolic subclass, exert extensive in vitro anti-invasive and in vivo anti-metastatic activities." Cancer Metastasis Rev. [Epub ahead of print]
18.Islam, M. A. (2012). "Cardiovascular Effects of Green Tea Catechins: Progress and Promise." Recent Pat Cardiovasc Drug Discov. 7(2):88-99.
19.Kamiyama, M. and T. Shibamoto (2012). "Flavonoids with Potent Antioxidant Activity Found in Young Green Barley Leaves." J Agric Food Chem.60 (25): 6260−6267
20.Kong, J.-M., L.-S. Chia, et al. (2003). "Analysis and biological activities of anthocyanins." Phytochemistry 64(5): 923-933.
21.Konczak, I. and W. Zhang (2004). "Anthocyanins-More Than Nature's Colours." J Biomed Biotechnol 2004(5): 239-240.
22.Cooke, D., W. P. Steward, et al. (2005). "Anthocyans from fruits and vegetables – Does bright colour signal cancer chemopreventive activity?" European Journal of Cancer 41(13): 1931-1940.
23.Castañeda-Ovando, A., M. d. L. Pacheco-Hernández, et al. (2009). "Chemical studies of anthocyanins: A review." Food Chemistry 113(4): 859-871.
24.Stintzing, F. C. and R. Carle (2004). "Functional properties of anthocyanins and betalains in plants, food, and in human nutrition." Trends in Food Science & Technology 15(1): 19-38.
25.Steeg, P. S. (2003). "Metastasis suppressors alter the signal transduction of cancer cells." Nat Rev Cancer 3(1): 55-63.
26.Yapijakis, C., A. Bramos, et al. (2012). "The Interplay between Hemostasis and Malignancy: The Oral Cancer Paradigm." Anticancer Res 32(5): 1791-1800.
27.Wang, Y. and J. S. Lazo (2012). "Metastasis-associated phosphatase PRL-2 regulates tumor cell migration and invasion." Oncogene 31(7): 818-827.
28.Yadav, A., B. Kumar, et al. (2011). "IL-6 promotes head and neck tumor metastasis by inducing epithelial-mesenchymal transition via the JAK-STAT3-SNAIL signaling pathway." Mol Cancer Res 9(12): 1658-1667.
29.Steeg, P. S. (2003). "Metastasis suppressors alter the signal transduction of cancer cells." Nat Rev Cancer 3(1): 55-63.
30.Gross, J. and C. M. Lapiere (1962). "Collagenolytic activity in amphibian tissues: a tissue culture assay." Proc Natl Acad Sci U S A 48: 1014-1022.
31.Chandler, S., K. M. Miller, et al. (1997). "Matrix metalloproteinases, tumor necrosis factor and multiple sclerosis: an overview." J Neuroimmunol 72(2): 155-161.
32.Allan, J. A., A. J. Docherty, et al. (1995). "Binding of gelatinases A and B to type-I collagen and other matrix components." Biochem J 309 ( Pt 1): 299-306.
33.Suzuki, K., J. J. Enghild, et al. (1990). "Mechanisms of activation of tissue procollagenase by matrix metalloproteinase 3 (stromelysin)." Biochemistry 29(44): 10261-10270.
34.Rudek, M. A., J. Venitz, et al. (2002). "Matrix metalloproteinase inhibitors: do they have a place in anticancer therapy?" Pharmacotherapy 22(6): 705-720.
35.Perkins, N. D. (2012). "The diverse and complex roles of NF-kappaB subunits in cancer." Nat Rev Cancer 12(2): 121-132.
36.Mohamed, M. R. and G. McFadden (2009). "NFkB inhibitors: strategies from poxviruses." Cell Cycle 8(19): 3125-3132.
37.MTT【cite;http://www.biotek.com/resources/articles/quantification-cell-viability- epoch.html】
38.Transwell migration and invasion【cite;http://www.millipore.com/offices/cp3/tw】
39.Zhang, Y., S. K. Vareed, et al. (2005). "Human tumor cell growth inhibition by nontoxic anthocyanidins, the pigments in fruits and vegetables." Life Sci 76(13): 1465-1472.
40.Ji, B. C., W. H. Hsu, et al. (2009). "Gallic acid induces apoptosis via caspase-3 and mitochondrion-dependent pathways in vitro and suppresses lung xenograft tumor growth in vivo." J Agric Food Chem 57(16): 7596-7604.
41.Lai, K. C., A. C. Huang, et al. (2010). "Benzyl isothiocyanate (BITC) inhibits migration and invasion of human colon cancer HT29 cells by inhibiting matrix metalloproteinase-2/-9 and urokinase plasminogen (uPA) through PKC and MAPK signaling pathway." J Agric Food Chem 58(5): 2935-2942.
42.Ma, C. Y., W. T. Ji, et al. (2011). "Butein inhibits the migration and invasion of SK-HEP-1 human hepatocarcinoma cells through suppressing the ERK, JNK, p38, and uPA signaling multiple pathways." J Agric Food Chem 59(16): 9032-9038.
43.Chen, Y. Y., S. Y. Chiang, et al. (2010). "Emodin, aloe-emodin and rhein inhibit migration and invasion in human tongue cancer SCC-4 cells through the inhibition of gene expression of matrix metalloproteinase-9." Int J Oncol 36(5): 1113-1120.
44.Ling, H., H. Yang, et al. (2010). "6-Shogaol, an active constituent of ginger, inhibits breast cancer cell invasion by reducing matrix metalloproteinase-9 expression via blockade of nuclear factor-kappaB activation." Br J Pharmacol 161(8): 1763-1777.
45.Hseu, Y. C., C. R. Wu, et al. (2011). "Inhibitory effects of Physalis angulata on tumor metastasis and angiogenesis." J Ethnopharmacol 135(3): 762-771.
46.Lo, C., T. Y. Lai, et al. (2010). "Gallic acid induces apoptosis in A375.S2 human melanoma cells through caspase-dependent and -independent pathways." Int J Oncol 37(2): 377-385.
47.Huang, H. P., Y. W. Shih, et al. (2008). "Chemoinhibitory effect of mulberry anthocyanins on melanoma metastasis involved in the Ras/PI3K pathway." J Agric Food Chem 56(19): 9286-9293.
48.Sylvester, P. W. (2011). "Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability." Methods Mol Biol 716: 157-168.
49.Lamy, S., R. Lafleur, et al. (2007). "Anthocyanidins inhibit migration of glioblastoma cells: structure-activity relationship and involvement of the plasminolytic system." J Cell Biochem 100(1): 100-111.
50.Madhunapantula, S. V., P. J. Mosca, et al. (2011). "The Akt signaling pathway: An emerging therapeutic target in malignant melanoma." Cancer Biol Ther 12(12).
51.Karar, J. and A. Maity (2011). "PI3K/AKT/mTOR Pathway in Angiogenesis." Front Mol Neurosci 4: 51.
52.Vega, F. M., A. Colomba, et al. (2012). "RhoB regulates cell migration through altered focal adhesion dynamics." Open Biol 2(5): 120076.
53.Yoon, J. H., H. E. Kim, et al. (2012). "Caffeoylserotonin suppresses THP-1 monocyte adhesion and migration via inhibition of the integrin beta1/FAK/Akt signalling pathway." Fitoterapia. [Epub ahead of print]
54.Miller Tw Fau - Rexer, B. N., J. T. Rexer Bn Fau - Garrett, et al. "Mutations in the phosphatidylinositol 3-kinase pathway: role in tumor progression and therapeutic implications in breast cancer." (1465-542X (Electronic)).
55.Perkins, N. D. (2012). "The diverse and complex roles of NF-kappaB subunits in cancer." Nat Rev Cancer 12(2): 121-132.
56.Chambers, A. F., A. C. Groom, et al. (2002). "Dissemination and growth of cancer cells in metastatic sites." Nat Rev Cancer 2(8): 563-572.
57.Philip, S. and G. C. Kundu (2003). "Osteopontin induces nuclear factor kappa B-mediated promatrix metalloproteinase-2 activation through I kappa B alpha /IKK signaling pathways, and curcumin (diferulolylmethane) down-regulates these pathways." J Biol Chem 278(16): 14487-14497.
58.Harris, C. C. (2012). "Article withdrawal: Proanthocyanidins from grape seeds inhibit expression of matrix metalloproteinases in human prostate carcinoma cells, which is associated with the inhibition of activation of MAPK and NFkappaB." Carcinogenesis 33(5): 1121.
59.Tempesta, M. S. (2012). "Review of Plant Phenolics and Human Health: Biochemistry, Nutrition and Pharmacology Plant Phenolics and Human Health: Biochemistry, Nutrition and Pharmacology ." J Nat Prod 75(6): 1260.
60.Gupta, S. C., J. H. Kim, et al. (2010). "Regulation of survival, proliferation, invasion, angiogenesis, and metastasis of tumor cells through modulation of inflammatory pathways by nutraceuticals." Cancer Metastasis Rev 29(3): 405-434.
61.Han, C., H. Ding, et al. (2005). "Inhibition of the growth of premalignant and malignant human oral cell lines by extracts and components of black raspberries." Nutr Cancer 51(2): 207-217.
62.Zhang, Y., N. P. Seeram, et al. (2008). "Isolation and identification of strawberry phenolics with antioxidant and human cancer cell antiproliferative properties." J Agric Food Chem 56(3): 670-675.
63.Ghosh, D. and T. Konishi (2007). "Anthocyanins and anthocyanin-rich extracts: role in diabetes and eye function." Asia Pac J Clin Nutr 16(2): 200-208.
64.Yeh, S.-L., W.-Y. Wang, et al. (2006). "Flavonoids suppresses the enhancing effect of β-carotene on DNA damage induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A549 cells."Chemico-Biological Interactions 160(2): 175-182.
65.Kamiyama, M. and T. Shibamoto (2012). "Flavonoids with Potent Antioxidant Activity Found in Young Green Barley Leaves." J Agric Food Chem.
66.Mauray, A., C. Felgines, et al. (2012). "Bilberry anthocyanin-rich extract alters expression of genes related to atherosclerosis development in aorta of apo E-deficient mice." Nutrition, Metabolism and Cardiovascular Diseases 22(1): 72-80.
67.Ziberna, L., M. Lunder, et al. (2010). "Acute cardioprotective and cardiotoxic effects of bilberry anthocyanins in ischemia-reperfusion injury: beyond concentration-dependent antioxidant activity." Cardiovasc Toxicol 10(4): 283-294.
68.Li, J., S. S. Lim, et al. (2012). "Purple corn anthocyanins dampened high-glucose-induced mesangial fibrosis and inflammation: possible renoprotective role in diabetic nephropathy." The Journal of Nutritional Biochemistry 23(4): 320-331.
69.Islam, M. A. (2012). "Cardiovascular Effects of Green Tea Catechins: Progress and Promise." Recent Pat Cardiovasc Drug Discov7(2):88-99.
70.Mauray, A., C. Felgines, et al. (2010). "Nutrigenomic analysis of the protective effects of bilberry anthocyanin-rich extract in apo E-deficient mice." Genes Nutr 5(4): 343-353.
71.Han, K. H., A. Matsumoto, et al. (2007). "Effects of anthocyanin-rich purple potato flakes on antioxidant status in F344 rats fed a cholesterol-rich diet." Br J Nutr 98(5): 914-921.
72.Tanaka, J., S. Nakamura, et al. (2012). "Purple rice (Oryza sativa L.) extract and its constituents inhibit VEGF-induced angiogenesis." Phytother Res 26(2): 214-222.
73.Chen, P. N., W. H. Kuo, et al. (2006). "Black rice anthocyanins inhibit cancer cells invasion via repressions of MMPs and u-PA expression." Chem Biol Interact 163(3): 218-229.
74.Olsson, M. E., C. S. Andersson, et al. (2006). "Antioxidant levels and inhibition of cancer cell proliferation in vitro by extracts from organically and conventionally cultivated strawberries." J Agric Food Chem 54(4): 1248-1255.
75.Ding, M., R. Feng, et al. (2006). "Cyanidin-3-glucoside, a natural product derived from blackberry, exhibits chemopreventive and chemotherapeutic activity." J Biol Chem 281(25): 17359-17368.
76.Zu, X. Y., Z. Y. Zhang, et al. (2010). "Anthocyanins extracted from Chinese blueberry (Vaccinium uliginosum L.) and its anticancer effects on DLD-1 and COLO205 cells." Chin Med J (Engl) 123(19): 2714-2719.
77.Seeram, N. P., L. S. Adams, et al. (2004). "Total cranberry extract versus its phytochemical constituents: antiproliferative and synergistic effects against human tumor cell lines." J Agric Food Chem 52(9): 2512-2517.
78.Seeram, N. P., L. S. Adams, et al. (2006). "Blackberry, black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts inhibit growth and stimulate apoptosis of human cancer cells in vitro." J Agric Food Chem 54(25): 9329-9339.
79.Wang, Y. and J. S. Lazo (2012). "Metastasis-associated phosphatase PRL-2 regulates tumor cell migration and invasion." Oncogene 31(7): 818-827.
80.Hseu, Y. C., C. R. Wu, et al. (2011). "Inhibitory effects of Physalis angulata on tumor metastasis and angiogenesis." J Ethnopharmacol 135(3): 762-771.
81.Ho, Y. T., J. S. Yang, et al. (2009). "Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-kappaB, u-PA and MMP-2 and -9." Cancer Lett 279(2): 155-162.
82.Karar, J. and A. Maity (2011). "PI3K/AKT/mTOR Pathway in Angiogenesis." Front Mol Neurosci 4: 51.
83.Madhunapantula, S. V., P. J. Mosca, et al. (2011). "The Akt signaling pathway: An emerging therapeutic target in malignant melanoma." Cancer Biol Ther 12(12).
84.Adams, L. S., S. Phung, et al. (2010). "Blueberry phytochemicals inhibit growth and metastatic potential of MDA-MB-231 breast cancer cells through modulation of the phosphatidylinositol 3-kinase pathway." Cancer Res 70(9): 3594-3605.
85.Huang, C.-S., Y.-E. Fan, et al. (2007). "Lycopene inhibits matrix metalloproteinase-9 expression and down-regulates the binding activity of nuclear factor-kappa B and stimulatory protein-1." The Journal of Nutritional Biochemistry 18(7): 449-456.
86.Prakash, M., S. Kale, et al. (2011). "Hyaluronan-binding protein 1 (HABP1/p32/gC1qR) induces melanoma cell migration and tumor growth by NF-kappa B dependent MMP-2 activation through integrin alpha(v)beta(3) interaction." Cell Signal 23(10): 1563-1577.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊