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dc.contributor.authorYea, His-Wenen_US
dc.identifier.citationAylon Y, Oren M (2007) Living with p53, dying of p53. Cell 130, 597-600. Bjorge JD, Jakymiw A, Fujita DJ (2000) Selected glimpses into the activation and function of Src kinase. Oncogene 19, 5620-35. Brown MC, Perrotta JA, Turner CE (1996) Identification of LIM3 as the principal determinant of paxillin focal adhesion localization and characterization of a novel motif on paxillin directing vinculin and focal adhesion kinase binding. J Cell Biol 135, 1109-23. Brown MT, Cooper JA (1996) Regulation, substrates and functions of src. Biochim Biophys Acta 1287, 121-49. Chen HC, Chan PC, Tang MJ, Cheng CH, Chang TJ (1998) Tyrosine phosphorylation of focal adhesion kinase stimulated by hepatocyte growth factor leads to mitogen-activated protein kinase activation. J Biol Chem 273, 25777-82. Chen W, Possemato R, Campbell KT, Plattner CA, Pallas DC, Hahn WC (2004) Identification of specific PP2A complexes involved in human cell transformation. Cancer Cell 5, 127-36. Coe BP, Henderson LJ, Garnis C, Tsao MS, Gazdar AF, Minna J, Lam S, Macaulay C, Lam WL (2005) High-resolution chromosome arm 5p array CGH analysis of small cell lung carcinoma cell lines. Genes Chromosomes Cancer 42, 308-13. Condeelis J, Segall JE (2003) Intravital imaging of cell movement in tumours. Nat Rev Cancer 3, 921-30. Deng X, Ito T, Carr B, Mumby M, May WS, Jr. (1998) Reversible phosphorylation of Bcl2 following interleukin 3 or bryostatin 1 is mediated by direct interaction with protein phosphatase 2A. J Biol Chem 273, 34157-63. Deryugina EI, Quigley JP (2006) Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 25, 9-34. Deryugina EI, Zijlstra A, Partridge JJ, Kupriyanova TA, Madsen MA, Papagiannakopoulos T, Quigley JP (2005) Unexpected effect of matrix metalloproteinase down-regulation on vascular intravasation and metastasis of human fibrosarcoma cells selected in vivo for high rates of dissemination. Cancer Res 65, 10959-69. Eichhorn PJ, Creyghton MP, Wilhelmsen K, van Dam H, Bernards R (2007) A RNA interference screen identifies the protein phosphatase 2A subunit PR55gamma as a stress-sensitive inhibitor of c-SRC. PLoS Genet 3, e218. Fomenko E, Slepov EV, Inzhevatkin EV, Savchenko AA (2006) [Bioluminescent method for determination of concentration metabolic substrates and NAD+ in the lymphocytes]. Biomed Khim 52, 507-10. Fontanini G, Vignati S, Bigini D, Mussi A, Lucchi M, Angeletti CA, Basolo F, Bevilacqua G (1995) Bcl-2 protein: a prognostic factor inversely correlated to p53 in non-small-cell lung cancer. Br J Cancer 71, 1003-7. Friedl P, Wolf K (2003) Proteolytic and non-proteolytic migration of tumour cells and leucocytes. Biochem Soc Symp, 277-85. Friedl P, Wolf K (2003) Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 3, 362-74. Garnis C, Baldwin C, Zhang L, Rosin MP, Lam WL (2003) Use of complete coverage array comparative genomic hybridization to define copy number alterations on chromosome 3p in oral squamous cell carcinomas. Cancer Res 63, 8582-5. Gotz J, Probst A, Ehler E, Hemmings B, Kues W (1998) Delayed embryonic lethality in mice lacking protein phosphatase 2A catalytic subunit Calpha. Proc Natl Acad Sci U S A 95, 12370-5. Guarino M, Rubino B, Ballabio G (2007) The role of epithelial-mesenchymal transition in cancer pathology. Pathology 39, 305-18. Guenin S, Schwartz L, Morvan D, Steyaert JM, Poignet A, Madelmont JC, Demidem A (2008) PP2A activity is controlled by methylation and regulates oncoprotein expression in melanoma cells: a mechanism which participates in growth inhibition induced by chloroethylnitrosourea treatment. Int J Oncol 32, 49-57. Gupta GP, Massague J (2006) Cancer metastasis: building a framework. Cell 127, 679-95. Hakem R, Hakem A, et al. (1998) Differential requirement for caspase 9 in apoptotic pathways in vivo. Cell 94, 339-52. He H, Jazdzewski K, et al. (2005) The role of microRNA genes in papillary thyroid carcinoma. Proc Natl Acad Sci U S A 102, 19075-80. He L, Thomson JM, et al. (2005) A microRNA polycistron as a potential human oncogene. Nature 435, 828-33. Hendrix P, Turowski P, Mayer-Jaekel RE, Goris J, Hofsteenge J, Merlevede W, Hemmings BA (1993) Analysis of subunit isoforms in protein phosphatase 2A holoenzymes from rabbit and Xenopus. J Biol Chem 268, 7330-7. Janssens V, Goris J (2001) Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem J 353, 417-39. Kamibayashi C, Estes R, Lickteig RL, Yang SI, Craft C, Mumby MC (1994) Comparison of heterotrimeric protein phosphatase 2A containing different B subunits. J Biol Chem 269, 20139-48. Kinoshita K, Nemoto T, Nabeshima K, Kondoh H, Niwa H, Yanagida M (1996) The regulatory subunits of fission yeast protein phosphatase 2A (PP2A) affect cell morphogenesis, cell wall synthesis and cytokinesis. Genes Cells 1, 29-45. Larue L, Bellacosa A (2005) Epithelial-mesenchymal transition in development and cancer: role of phosphatidylinositol 3'' kinase/AKT pathways. Oncogene 24, 7443-54. Le Sourd F, Boulben S, Le Bouffant R, Cormier P, Morales J, Belle R, Mulner-Lorillon O (2006) eEF1B: At the dawn of the 21st century. Biochim Biophys Acta 1759, 13-31. Li J (2007) Marker selection for whole-genome association studies with two-stage designs using dense single-nucleotide polymorphisms. BMC Proc 1 Suppl 1, S136. Li K, Li Y, Shelton JM, Richardson JA, Spencer E, Chen ZJ, Wang X, Williams RS (2000) Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell 101, 389-99. Li X, Yost HJ, Virshup DM, Seeling JM (2001) Protein phosphatase 2A and its B56 regulatory subunit inhibit Wnt signaling in Xenopus. Embo J 20, 4122-31. Liotta LA, Kohn EC (2001) The microenvironment of the tumour-host interface. Nature 411, 375-9. McCright B, Virshup DM (1995) Identification of a new family of protein phosphatase 2A regulatory subunits. J Biol Chem 270, 26123-8. Mimori K, Mori M, Inoue H, Ueo H, Mafune K, Akiyoshi T, Sugimachi K (1996) Elongation factor 1 gamma mRNA expression in oesophageal carcinoma. Gut 38, 66-70. Moreno CS, Park S, Nelson K, Ashby D, Hubalek F, Lane WS, Pallas DC (2000) WD40 repeat proteins striatin and S/G(2) nuclear autoantigen are members of a novel family of calmodulin-binding proteins that associate with protein phosphatase 2A. J Biol Chem 275, 5257-63. Mumby M (1995) Regulation by tumour antigens defines a role for PP2A in signal transduction. Semin Cancer Biol 6, 229-37. Mumby M (2007) The 3D structure of protein phosphatase 2A: new insights into a ubiquitous regulator of cell signaling. ACS Chem Biol 2, 99-103. Mumby M (2007) PP2A: unveiling a reluctant tumor suppressor. Cell 130, 21-4. Mumby MC, Walter G (1993) Protein serine/threonine phosphatases: structure, regulation, and functions in cell growth. Physiol Rev 73, 673-99. Nikkola J, Vihinen P, Vuoristo MS, Kellokumpu-Lehtinen P, Kahari VM, Pyrhonen S (2005) High serum levels of matrix metalloproteinase-9 and matrix metalloproteinase-1 are associated with rapid progression in patients with metastatic melanoma. Clin Cancer Res 11, 5158-66. Olarewaju O, Ortiz PA, Chowdhury WQ, Chatterjee I, Kinzy TG (2004) The translation elongation factor eEF1B plays a role in the oxidative stress response pathway. RNA Biol 1, 89-94. Panani AD, Ferti AD, Raptis SA, Roussos C (2004) Novel recurrent structural chromosomal aberrations in primary bladder cancer. Anticancer Res 24, 2967-74. Panani AD, Roussos C (2006) Cytogenetic and molecular aspects of lung cancer. Cancer Lett 239, 1-9. Rocha S, Nabais S, Magalhaes S, Salgado A, Azevedo P, Torres M, Marques J, Pereira MA, Correia A (2007) Heart failure after acute coronary syndrome: identify to treat better! Rev Port Cardiol 26, 349-59. Sandilands E, Cans C, Fincham VJ, Brunton VG, Mellor H, Prendergast GC, Norman JC, Superti-Furga G, Frame MC (2004) RhoB and actin polymerization coordinate Src activation with endosome-mediated delivery to the membrane. Dev Cell 7, 855-69. Santoro MF, Annand RR, et al. (1998) Regulation of protein phosphatase 2A activity by caspase-3 during apoptosis. J Biol Chem 273, 13119-28. Schild A, Schmidt K, Lim YA, Ke Y, Ittner LM, Hemmings BA, Gotz J (2006) Altered levels of PP2A regulatory B/PR55 isoforms indicate role in neuronal differentiation. Int J Dev Neurosci 24, 437-43. Sim AT (1991) The regulation and function of protein phosphatases in the brain. Mol Neurobiol 5, 229-46. Snaith HA, Armstrong CG, Guo Y, Kaiser K, Cohen PT (1996) Deficiency of protein phosphatase 2A uncouples the nuclear and centrosome cycles and prevents attachment of microtubules to the kinetochore in Drosophila microtubule star (mts) embryos. J Cell Sci 109 ( Pt 13), 3001-12. Sun Y, Yuan J, Liu H, Shi Z, Baker K, Vuori K, Wu J, Feng GS (2004) Role of Gab1 in UV-induced c-Jun NH2-terminal kinase activation and cell apoptosis. Mol Cell Biol 24, 1531-9. Tehrani MA, Mumby MC, Kamibayashi C (1996) Identification of a novel protein phosphatase 2A regulatory subunit highly expressed in muscle. J Biol Chem 271, 5164-70. Voorhoeve PM, Agami R (2007) Classifying microRNAs in cancer: the good, the bad and the ugly. Biochim Biophys Acta 1775, 274-82. Voorhoeve PM, Hijmans EM, Bernards R (1999) Functional interaction between a novel protein phosphatase 2A regulatory subunit, PR59, and the retinoblastoma-related p107 protein. Oncogene 18, 515-24. Voorhoeve PM, Watson RJ, Farlie PG, Bernards R, Lam EW (1999) Rapid dephosphorylation of p107 following UV irradiation. Oncogene 18, 679-88. Wera S, Fernandez A, Lamb NJ, Turowski P, Hemmings-Mieszczak M, Mayer-Jaekel RE, Hemmings BA (1995) Deregulation of translational control of the 65-kDa regulatory subunit (PR65 alpha) of protein phosphatase 2A leads to multinucleated cells. J Biol Chem 270, 21374-81. Wikenheiser-Brokamp KA (2006) Retinoblastoma regulatory pathway in lung cancer. Curr Mol Med 6, 783-93. Wistuba, II (2007) Genetics of preneoplasia: lessons from lung cancer. Curr Mol Med 7, 3-14. Wistuba J, Mittag J, Luetjens CM, Cooper TG, Yeung CH, Nieschlag E, Bauer K (2007) Male congenital hypothyroid Pax8-/- mice are infertile despite adequate treatment with thyroid hormone. J Endocrinol 192, 99-109. Yokoyama N, Miller WT (2001) Inhibition of Src by direct interaction with protein phosphatase 2A. FEBS Lett 505, 460-4. Yokoyama N, Miller WT (2001) Protein phosphatase 2A interacts with the Src kinase substrate p130(CAS). Oncogene 20, 6057-65.en_US
dc.description.abstract近年來,台灣的肺癌病例有顯著增加的趨勢,在女性的癌症病例中因肺癌而死亡的人數已高居首位。因此了解腫瘤生成及轉移的機制成為當務之急。進年來在台灣,肺癌早已躍居女性癌症死亡率第一位,因此為了找出肺腺癌轉移及侵入的機制是非常重要的。先前利用微陣列技術以及微陣列-比較基因組雜交技術,交叉分析肺腺癌侵入/轉移細胞株 CL1-0和CL1-5,篩選出可能與腫瘤轉移相關的基因。結果篩選出一個功能仍未被完全定義的基因PPP2R2C。該基因隸屬於PP2A調控次單元的其中一員,但因該基因的功能仍不確定,因此我們計劃將pEF6/V5-PPP2R2C送入CL1-5細胞株中,觀察當大量表現PPP2R2C後,會大幅降低CL1-5的侵入、移動、增生以及細胞聚落形成的能力。蛋白質交互作用為細胞中蛋白質執行功能的重要環節,一方面利用免疫沉澱法,發現細胞內生性的c-SRC會與PPP2R2C有交互作用。而大量表現PPP2R2C會抑制c-SRC活化以及Ras-ERK1/2的磷酸化程度。另一方面,利用GST 沉降試驗來尋找其他可能與PPP2R2C有交互作用的蛋白。由質譜分析結果顯示,eukaryotic translation elongation factor 1 gamma (EEF1G)為參與細胞內tRNA的運送,可能會與PPP2R2C有交互作用。總合以上的結果PPP2R2C可能參與調控細胞的侵犯以及轉移機制,本研究提供更多明確的方向供後續研究,期望能發展新的抗癌治療標的。zh_TW
dc.description.abstractRecently, lung cancer has dominantly increased and ranked up to the first place among Taiwanese women died from cancers. Therefore, understanding the tumorigenesis and metastasis in lung cancer is very important. Previously we have identified ppp2r2c, a gene for regulatory subunit of the PP2A complex, by using microarray and array-CGH in lung cancer cell lines that is with various invasive abilities. In order to clarify the role of PPP2R2C in NSCLC, we stably transfect human CL1-5 lung cancer cells with pEF6/V5-PPP2R2C and found decreased invasive activity in the resulting clones compared to control cells. In addition, compare to control cell, cell that overexpresses PPP2R2C decreased proliferation and colony formation rate in soft agar. To address the celluar function of PPP2R2C, we examined the known biochemical regulators of invasion and proliferation. In immunoprecipitation experiment, endogenous c-SRC co-immunoprecipitate with PPP2R2C-V5. Additionally, overexpression PPP2R2C decreased c-SRC activityand markedly suppressed the activation of Ras-ERK1/2 in highly invasive CL1-5. On the other hand, we have identified the interacting proteins of PPP2R2C through affinity purification-based proteomics to sort out the candidate proteins. The result revealed that eukaryotic translation elongation factor 1 gamma (EEF1G) could interact with PPP2R2C. EEF1G encodes a subunit of the elongation factor-1 complex which's function is known as enzymatic delivery of aminoacyl tRNAs to the ribosome. This study gives us a new annotation of PPP2R2C in cancer metastasis and invasion mechanism and that PPP2R2C might be a promising therapeutic target for cancer metastasis and cell growth.en_US
dc.description.tableofcontents第一章 序論 第一節 前人研究 1 一、肺癌 1 二、腫瘤細胞之生成 2 三、癌細胞轉移 3 四、比較型基因體雜交分析技術 5 五、蛋白質磷酸化 7 六、PP2A結構及次單元 7 七、PP2A的調節功能 8 八、c-SRC酪氨酸激酶家族之結構及功能 9 九、c-Src酪氨酸激酶的調控機制 10 第二節 研究目的和策略架構 11 第二章 材料與方法 第一節 細胞培養 13 第二節 細胞冷凍保存及解凍 13 第三節 構築載體 13 第四節 勝任細胞製備 14 第五節 轉型作用 14 第六節 細菌質體DNA抽取 15 第七節 細胞染色體DNA萃取 15 第八節 共轉染作用 16 第九節 細胞RNA萃取 16 第十節 即時定量反轉錄聚合酶連鎖反應 17 第十一節 細胞總蛋白質抽取 18 第十二節 蛋白質定量 19 第十三節 蛋白質凝膠電泳分析 19 第十四節 西方墨點法 20 第十五節 免疫螢光染色及纖維型肌動蛋白(F-actin)染色 21 第十六節 細胞基質侵襲力分析(matrixgel gel invasion) 21 第十七節 細胞遷移分析(cell migration assay) 22 第十八節 細胞增生分析 22 第十九節 細胞非貼附性分析 23 第二十節 純化GST-PPP2R2C蛋白 23 第二十一節 GST-pull down 分析 24 第二十二節 分離萃取核質蛋白 24 第三章 結果 26 第一節 PPP2R2C在mRNA表現量以及蛋白表現量在CL1-0高於CL1-5 26 第二節 PPP2R2C基因構築 26 第三節 PPP2R2C進行穩定表現細胞株加藥篩選 26 第四節 穩定大量表現PPP2R2C會改變CL1-5細胞型態並降低細胞移動以及侵犯能力 27 第五節 穩定大量表現PPP2R2C會降低CL1-5細胞增生以及貼附能力 28 第六節 穩定大量表現PPP2R2C會降低CL1-5非貼附生長能力 28 第七節 PPP2R2C蛋白主要在細胞質中表現 28 第八節 以GST沉澱分析與PPP2R2C有交互作用的蛋白 29 第九節 c-SRC與PPP2R2C有交互作用 29 第十節 大量表現PPP2R2C會抑制CL1-5內生性SRC以及ERK的磷酸化程度 30 第十一節 EEF1G與PPP2R2C有交互作用 31 第四章 討論 32 第五章 結論 36 參考文獻 37 實驗結果圖表 43 附表及附圖 68 附錄 80zh_TW
dc.subjectSRC kinaseen_US
dc.titleThe functional analysis of PPP2R2C in non-small-cell lung carcinomaen_US
dc.typeThesis and Dissertationzh_TW
item.openairetypeThesis and Dissertation-
item.fulltextno fulltext-
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