Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/81163
DC FieldValueLanguage
dc.contributor闕斌如zh_TW
dc.contributorPin-Ju Chuehen_US
dc.contributor.author呂卓穎zh_TW
dc.contributor.authorLu, Cho-Yingen_US
dc.contributor.other生命科學院碩士在職專班zh_TW
dc.date2013en_US
dc.date.accessioned2014-06-13T08:11:07Z-
dc.date.available2014-06-13T08:11:07Z-
dc.identifierU0005-0608201320442100en_US
dc.identifier.citation1.Aita V.M., Liang X.H., Murty V.V., Pincus D.L., Yu W., Cayanis E., Kalachikov S., Gilliam T.C., Levine B.: Cloning and genomic organization of beclin 1, a candidate tumor suppressor gene on chromosome 17q21. Genomics 1999, 59:59-65. 2.Athanasiou A., Smith P.A., Vakilpour S., Kumaran N.M., Turner A.E., Bagiokou D. et al. (2007). Vanilloid receptor agonists and antagonists are mitochondrial inhibitors: how vanilloids cause non-vanilloid receptor mediated cell death. Biochem Biophys Res Commun 354: 50–55. 3.Brozmanova, J.; Manikova, D.; Vlckova, V.; Chovanec, M. Selenium: A double-edged sword for defense and offence in cancer. Arch. Toxicol. 2010, 84, 919–938. 4.Chen N., Debnath J.: Autophagy and tumorigenesis. FEBS Lett 2010, 584:1427-1435. 5.Crighton D., Wilkinson S., Ryan K.M. DRAM links autophagy to p53 and programmed cell death. Autophagy. 2007;3(1):72–74. 6.Debnath, J., E.H. Baehrecke, G. Kroemer. 2005. Does autophagy contribute to cell death? Autophagy 1, 66–74. 7.Dunn Jr. W.A., Cregg J.M., Kiel J.A.K.W., van der Klei I.J., Oku M., Sakai Y., Sibirny A.A., Stasyk O.V., Veenhuis M. Pexophagy: The selective autophagy of peroxisomes. Autophagy 2005; 1:75:83. 8.Fimia G.M., Stoykova A., Romagnoli A., Giunta L., Di Bartolomeo S., Nardacci R., Corazzi M., Fuoko C., Ucar A., Schwartz P. et al.: Ambra 1 regulates autophagy and development of the nervous system. Nature 2007, 447:1121-1125. 9.Finn P.F., Dice J.F. Ketone bodies stimulate chaperone mediated autophagy. J Biol Chem 2005; 280:25864:70. 10.Fung C., Lock R., Gao S., Salas E., Debnath J: Induction of autophagy during extracellular matrix detachment promotes cell survival. Mol Biol Cell 2008, 19:797-806. 11.Hosokawa N., Hara Y., Mizushima N. Generation of cell lines with tetracycline regulated autophagy and a role for autophagy in controlling cell size. FEBS Lett 2006; 580:2623:9. 12.Ito K., Nakazato T., Yamato K., Miyakawa Y., Yamada T., Hozumi N. et al. (2004). Induction of apoptosis in leukemic cells by homovanillic acid derivative, capsaicin, through oxidative stress: implication of phosphorylation of p53 at Ser-15 residue by reactive oxygen species. Cancer Res 64: 1071–1078. 13.Jerant A.F., Johnson J.T., Sheridan C.D., Caffrey T.J. (July 2000). Early detection and treatment of skin cancer. Am Fam Physician 62 (2): 357–68, 375–6, 381–2. 14.Jung M.Y., Kang H.J., Moon A. (2001). Capsaicin-induced apoptosis in SK-Hep-1 hepatocarcinoma cells involves Bcl-2 downregulation and capsase-3 activation. Cancer Lett 165: 139–145. 15.Kim C.S., Park W.H., Park J.Y., Kang J.H., Kim M.O., Kawada T. et al. (2004). Capsaicin, a spicy component of hot pepper, induces apoptosis by activation of the peroxisome proliferator-activated receptor gamma in HT-29 human colon cancer cells. J Med Food 7: 267–273. 16.Kuma A., Hatano M., Matsui M., Yamamoto A., Nakaya H., Yoshimori T., Ohsumi Y., Tokuhisa T., Mizushima N. The role of autophagy during the early neonatal starvation period. Nature 2004; 432:1032:6. 17.Levine, B., D. J. Klionsky. 2004. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell 6, 463–477. 18.Li B.X., Li C.Y., Peng R.Q., et al. The expression of beclin 1 is associated with favorable prognosis in stage IIIB colon cancers. Autophagy. 2009;5(3):303–306. 19.Li L., Lou Z., Wang L.: The role of FKBP5 in cancer aetiology and chemoresistance. Br J Cancer 2011, 104:19-23. 41. 20.Li L., Fridley B., Kalari K., Jenkins G., Batzler A., Safgren S., Hildebrandt M., Ames M., Schaid D., Wang L.: Gemcitabine and cytosine arabinoside cytotoxicity: association with lymphoblastoid cell expression. Cancer Res 2008, 68:7050-7058. 21.Liang X.H., Jackson S., Seaman M., Brown K., Kempkes B., Hibshoosh H., Levine B.: Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 1999, 402:672-676. 22.Lippman, S.M.; Klein, E.A.; Goodman, P.J.; Lucia, M.S.; Thompson, I.M.; Ford, L.G.; Parnes, H.L.; Minasian, L.M.; Gaziano, J.M.; Hartline, J.A.; et al. Effect of selenium and vitamin E on risk of prostate cancer and other cancers: The selenium and vitamin E cancer prevention trial (select). JAMA 2009, 301, 39–51. 23.Lo Y.C., Yang Y.C., Wu I.C., Kuo F.C., Liu C.M., Wang H.W. et al. (2005). Capsaicin-induced cell death in a human gastric adenocarcinoma cell line. World J Gastroenterol 11: 6254–6257. 24.Lucas, Robyn; McMichael, Tony; Smith, Wayne; Armstrong, Bruce (2006). Solar Ultraviolet Radiation: Global burden of disease from solar ultraviolet radiation. Environmental Burden of Disease Series 13. World Health Organization. 25.Ma X., Piao S., Wang D.W., McAfee Q.W., Nathanson K.L., Lum J.J., Li L.Z., Amaravadi R: Measurements of tumor cell autophagy predict invasiveness, resistance to chemotherapy, and survival in melanoma. Clin Cancer Res 2011. 26.Maiuri M.C., Le Toumelin G., Criollo A., Rain J.C., Gautier F., Juin P., Tasdemir E., Pierron G., Troulinaki K., Tavernarakis N. et al.: Functional and physical interaction between Bcl-X(L) and a BH3-like domain in Beclin-1. EMBO J 2007, 26:2527-2539. 27.Mathew R., Karp C.M., Beaudoin B., Vuong N., Chen G., Chen H.Y., Bray K., Reddy A., Bhanot G., Gelinas C. et al.: Autophagy suppresses tumorigenesis through elimination of p62. Cell 2009, 137:1062-1075. 28.Mathew R., Karantza-Wadsworth V., White E. Role of autophagy in cancer. Nat Rev Cancer. 2007;7(12):961–967. 29.Mizushima N.: The role of the Atg1/ULK1 complex in autophagy regulation. Curr Opin Cell Biol 2010, 22:132-139. 30.Mori A., Lehmann S., O’Kelly J., Kumagai T., Desmond JC., Pervan M. et al. (2006). Capsaicin, a component of red peppers, inhibits the growth of androgen-independent, p53 mutant prostate cancer cells. Cancer Res 66: 3222–3229. 31.Morselli E., Galluzzi L., Kepp O., Vicencio J.M., Criollo A., Maiuri M.C., Kroemer G.: Anti- and pro-tumor functions of autophagy. Biochim Biophys Acta 2009, 1793:1524-1532. 32.Morishima N., Nakanishi K., Takenouchi H., Shibata T., and Yasuhiko Y., “An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12,” Journal of Biological Chemistry, vol. 277, no. 37, pp. 34287–34294, 2002. 33.Nakagawa T., Zhu H., Morishima N. et al., “Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-β,” Nature, vol. 403, no. 6765, pp. 98–103, 2000. 34.Pattingre S., Tassa A., Qu X., Garuti R., Liang X.H., Mizushima N., Packer M., Schneider M.D., Levine B.: Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 2005, 122:927-939. 35.Rao R. V., Castro-Obregon S., Frankowski H. et al., “Coupling endoplasmic reticulumstress to the cell death program. An Apaf-1-independent intrinsic pathway,” Journal of Biological Chemistry, vol. 277, no. 24, pp. 21836–21842, 2002. 36.Ravikumar B., Futter M., Jahreiss L., Korolchuk V.I., Lichtenberg M., Luo S., Massey D.C., Menzies F.M., Narayanan U., Renna M. et al.: Mammalian macroautophagy at a glance. J Cell Sci 2009, 122:1707-1711 37.Romano M.F., Avellino R., Petrella A., Bisogni R., Romano S., Venuta S.: Rapamycin inhibits doxorubicin-induced NFkappaB/ Rel nuclear activity and enhances the apoptosis of melanoma cells. Eur J Cancer 2004, 40:2829-2836. 38.Romano S., D’Angelillo A., Pacelli R., Staibano S., De Luna E., Bisogni R., Eskelinen E.L., Mascolo M., Cali G., Arra C. et al.: Role of FK506-binding protein 51 in the control of apoptosis of irradiated melanoma cells. Cell Death Differ 2010, 17:145-157. 39.Sa′ nchez A.M., Sa′ nchez M.G., Malagarie-Cazenave S., Olea N., Dı′az-Laviada I. (2006). Induction of apoptosis in prostate tumor PC-3 cells and inhibition of xenograft prostate tumor growth by the vanilloid capsaicin. Apoptosis 11: 89–99. 40.Shin D.H., Kim O.H., Jun H.S., Kang M.K., Inhibitory effect of capsaicin on B16-F10 melanoma cell migration via the phosphatidylinositol 3-kinase /Akt/ Rac1 signal pathway. Exp Mol Med. 2008;40:486–494. 41.Takahashi Y., Coppola D., Matsushita N., Cualing H.D., Sun M., Sato Y., Liang C., Jung J.U., Cheng J.Q., Mul J.J. et al.: Bif-1 interacts with Beclin 1 through UVRAG and regulates autophagy and tumorigenesis. Nat Cell Biol 2007, 9:1142-1151. 42.Terlecky S.R., Dice J.F. Polypeptide import and degradation by isolated lysosomes. J Biol Chem 1993; 268:23490‑5. 43.Tewksbury, J. J.; Nabhan, G. P. (2001). Seed dispersal. Directed deterrence by capsaicin in chilies. Nature 412 (6845): 403–404. 44.Wu, M.; Kang, M.M.; Schoene, N.W.; Cheng, W.H. Selenium compounds activate early barriers of tumorigenesis. J. Biol. Chem. 2010, 285, 12055–12062. 45.Yang S, Wang X, Contino G., Liesa M., Sahin E., Ying H., Bause A., Li Y., Stommel J.M., Dell’antonio G. et al.: Pancreatic cancers require autophagy for tumor growth. Genes Dev 2011. 46.Yee K.S., Wilkinson S., James J., Ryan K.M., Vousden K.H. PUMA- and Bax-induced autophagy contributes to apoptosis. Cell Death Differ. 2009;16(8):1135–1145. 47.Yoneda T., Imaizumi K., Oono K. et al., “Activation of caspase-12, an endoplastic reticulum (ER) resident caspase, through tumor necrosis factor receptor-associated factor 2- dependent mechanism in response to the ER stress,” Journal of Biological Chemistry, vol. 276, no. 17, pp. 13935–13940, 2001. 48.Yoshimori, T. 2004. Autophagy: a regulated bulk degradation process inside cells. Biochem Biophys Res Commun 313, 453–458. 49.Yue Z., Jin S., Yang C., Levine A.J., Heintz N.: Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci USA 2003, 100:15077-15082. 50.Zeng, H. Selenium as an essential micronutrient: Roles in cell cycle and apoptosis. Molecules 2009, 14, 1263–1278. 51.Zeng, H.; Combs, G.F., Jr. Selenium as an anticancer nutrient: Roles in cell proliferation and tumor cell invasion. J. Nutr. Biochem. 2008, 19, 1–7.en_US
dc.identifier.urihttp://hdl.handle.net/11455/81163-
dc.description.abstractMelanoma is a malignant tumor of melanocytes. If melanoma is found at early stage, it can be removed completely by surgery and the chance of cure is high. Unfortunately, if it is diagnosed late it may lead to skin cancer-relatred death. The treatments to melanoma these days include chemo- and immunotherapy, and radiation therapy. Capsaicin has an anti-proliferative effect in vitro on prostate, colon, gastric, hepatic and leukemic cancer cell lines. It is demonstrated that capsaicin inhibited melanoma cacncer cell lines, however, most of the mechanisms are still unclear. In this study, we used rat melanoma cell line, B16F10, for the evaluation of the anticancer effect of capsaicin . We found that the percentage of apoptosis and changes of mitochondrial membrane potential are notably increased in high concentration of capsaicin (400 μM). Western blot also shows that high concentration of capsaicin decreases Bcl-2 protein. Low concentrations of capsaicin are found to not to activate PARP (Poly ADP ribose polymerase) resulting in no apoptosis. While oxidative stress is not significantly different among different concentrations of capsaicin. We suggest that high concentration of capsaicin might induce apoptosis in B16F10 through mitochondrial dependent pathway. We also found that low concentrations of capsaicin (10, 100 μM) result in autophagy induction in B16F10 cells. Western blot also shows that high concentration of capsaicin increases the level of mTOR phosphorylation. On the other hand, low concentrations of capsaicin increase beclin-1 and autophagy. We suggest that capsaicin-mediated autophagy may involved in cell survival mechanism in B16F10 cells. Thus, we propose to utilize inhibitors of autophagy to examine the percentage of apoposis in B16F10. Similarly, inhibitors of apoptosis will be used to study the autophagy mechanism in B16F10 cells, with the hope to find out the relationship between autophagy and apoptosis in capsaicin-exposed B16F10 cells.en_US
dc.description.tableofcontents中文摘要 …………………………………………………………… i 英文摘要 …………………………………………………………… iii 目錄 ………………………………………………………………… v 圖表目錄…………………………………………………………… vii 壹、緒論…………………………………………………………… 1 一、辣椒素 (Capsaicin) …………………………………… 1 二、黑色素瘤細胞 (Melanoma) …………………………… 1 三、細胞凋亡 (Apoptosis)………………………………… 2 四、細胞自噬 (Autophagy) ………………………………… 5 貳、實驗材料……………………………………………………… 9 參、實驗方法 ……………………………………………………… 13 一、細胞培養 (Cell culture) …………………………… 13 二、西方墨漬蛋白表現偵測 (Western blotting) ……… 13 三、細胞凋亡測試 (Apoptosis assay) …………………… 15 四、過氧化物含量分析 (ROS measurement ) …………… 15 五、細胞自噬測試 (Autophagy assay)…………………… 16 六、粒線體膜通透性測試 (Mitochondrial membrane potential assay) 16 七、細胞數計數 (Trypan Blue Assay) …………………… 17 八、xCELLgence System儀分析 (xCELLgence System) … 17 九、統計分析 (Staistical analysis) …………………… 18 肆、實驗結果 ……………………………………………………… 19 一、辣椒素具有抑制B16F10細胞生長的效果……………… 19 二、辣椒素主要引發粒線體路徑的細胞凋亡……………… 20 三、辣椒素在較低濃度會引發細胞自噬…………………… 20 (圖一) B16F10在辣椒素作用下的細胞生長曲線………………… 22 (圖二) B16F10在辣椒素作用下的細胞計數……………………… 23 (圖三) B16F10在辣椒素作用下的細胞凋亡偵測………………… 24 (圖四) B16F10在辣椒素作用下的細胞凋亡偵測 (統計圖表) … 25 (圖五) B16F10在辣椒素作用下的西方墨點法偵測 (細胞凋亡代表性蛋白) 26 (圖六) B16F10在辣椒素作用下的粒線體膜通透性測試………… 27 (圖七) B16F10在辣椒素作用下的超氧化物 (ROS)測試………… 28 (圖八) B16F10在辣椒素作用下的細胞自噬測試………………… 29 (圖九) B16F10在辣椒素作用下的西方墨點法偵測 (細胞自噬代表性蛋白) 30 (表一) B16F10在辣椒素作用下的西方墨點法偵測 (細胞自噬代表性蛋白)數據化 31 伍、討論 …………………………………………………………… 32 陸、參考資料 ……………………………………………………… 34zh_TW
dc.language.isozh_TWen_US
dc.publisher生命科學院碩士在職專班zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-0608201320442100en_US
dc.subject辣椒素zh_TW
dc.subjectcapsaicinen_US
dc.subject細胞自噬zh_TW
dc.subject細胞凋亡zh_TW
dc.subject黑色素瘤zh_TW
dc.subjectautophagyen_US
dc.subjectapoptosisen_US
dc.subjectmelanomaen_US
dc.title辣椒素在 B16F10 細胞株引發細胞自噬及細胞凋亡zh_TW
dc.titleCapsaicin induces apoptosis and autophagy in B16F10 cellsen_US
dc.typeThesis and Dissertationzh_TW
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.fulltextno fulltext-
item.languageiso639-1zh_TW-
item.grantfulltextnone-
Appears in Collections:生命科學院
Show simple item record
 
TAIR Related Article

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.