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博碩士論文 etd-0611118-123519 詳細資訊
Title page for etd-0611118-123519
論文名稱
Title
EMP2 基因在膀胱尿路上皮癌細胞中透過 TGFB-SP1 路徑以及 P2RX7 的結合而抑制細胞增殖並誘導細胞凋亡
Epithelial membrane protein 2 inhibits cell proliferation and induces apoptosis via TGFB-SP1 axis and recruitment of P2RX7 in urinary bladder urothelial carcinoma-derived cells
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
72
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2018-07-09
繳交日期
Date of Submission
2018-07-11
關鍵字
Keywords
細胞週期、TGFB、SP1 (Specificity protein-1)、膀胱癌、Epithelial membrane protein 2 (EMP2)
SP1 (Specificity protein-1), TGFB, Epithelial membrane protein 2 (EMP2), Urinary bladder urothelial carcinoma (UBUC), Cell cycle arrest
統計
Statistics
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中文摘要
在我們的研究中,實驗目的為探討 EMP2 基因在膀胱尿路上皮癌細胞中,抑制細胞增殖並誘導細胞凋亡的機制。首先,我們進行一連串的體外測定,顯示了在膀胱癌細胞中,EMP2 會調節 TGFB2 以及誘導 SMAD2/3 的活化。而在本篇研究中,我們表明通過調節細胞週期蛋白依賴性激酶抑製劑 (Cyclin-dependent kinase inhibitors, CKIs),能夠使 EMP2/TGFB 和 PI3K/AKT 信號途徑之間產生相互作用。我們發現經由 TGFB 信號傳導途徑,外源表達 EMP2 能夠上調 CDKN1A,CDKN1B,CDKN1C 和 CDKN2B 的蛋白質表現。接下來藉由 J82 細胞中的 TGFB/SP1 信號傳遞途徑,當表達 EMP2 基因的外源性時,能夠活化上調 CDKN1A,CDKN1B 和 CDKN1C 的蛋白質表現。除此之外,我們利用 pull-down 測定,透過使用 DDK 抗體以及 P2RX7 抗體進行西方墨點法,證明了 EMP2 與 P2RX7 之間的交互作用。因此在 J82 細胞中,EMP2 通過與 P2RX7 相互作用誘導細胞外在和內在的凋亡。綜合以上,我們目前的研究已經證實了 EMP2 基因經由 TGFB/SP1 信號傳遞途徑的激活,參與調控細胞週期的影響。除此之外,我們的研究也指出 EMP2 通過與 P2RX7 的結合,抑制細胞增殖並誘導細胞凋亡。
Abstract
In this study, our objective was to investigate the mechanisms of Epithelial membrane protein 2 (EMP2) inhibits cell proliferation and induces apoptosis in urinary bladder urothelial carcinoma-derived cells. A series of in vitro assays indicated that EMP2 regulates transforming growth factor beta 2 (TGFB2) and induces SMAD2/3 activation in bladder cancer cells. Herein, we show that signaling interplay between EMP2/TGFB and PI3K/AKT pathway in the regulation of several cyclin-dependent kinase inhibitors (CKIs). We showed that exogenous EMP2 expression upregulated CDKN1A, CDKN1B, CDKN1C and CDKN2B protein levels via the TGFB signaling pathway. Afterward, exogenous expression of the EMP2 gene upregulated CDKN1A, CDKN1B and CDKN1C protein levels through TGFB/SP1 activation in J82 cells. In addition, pull-down assay using anti-DDK antibody and immunoblotting with anti-P2RX7 antibody demonstrated that EMP2 interacted with the P2RX7 protein. Therefore, EMP2 induced both extrinsic and intrinsic apoptosis through interaction with P2RX7 protein in J82 cells. Combined, the current studies have identified EMP2 gene via TGFB/SP1 activation, and that involved in the regulation of cell cycle. Apart from this, our study indicate EMP2 inhibits cell proliferation and induces apoptosis through recruitment of P2RX7 protein.
目次 Table of Contents
論文審定書 ⅰ
致謝 ⅱ
中文摘要 ⅲ
英文摘要 ⅳ
英文縮寫表 ⅵ
圖次 ⅶ
表次 ⅷ
壹、 緒論 (Introduction) 1
貳、 實驗材料與方法 (Materials and methods) 7
參、 結果 (Results) 24
肆、 討論 (Discussion) 54
伍、 參考文獻 (References) 57
陸、 Supplement Data 60
參考文獻 References
American Cancer Society. Bladder cancer. Atlanta, GA., 2018.
Abdelrahim, M., Smith, R., Burghardt, R.,Safe, S. (2004). Role of Sp proteins in regulation of vascular endothelial growth factor expression and proliferation of pancreatic cancer cell. cancer research.
Anumanthan, G., Halder, S.K., Osada, H., Takahashi, T., Massion, P.P., Carbone, D.P.,Datta, P.K. (2005). Restoration of TGF-beta signalling reduces tumorigenicity in human lung cancer cells. British J of Cancer.
Attardi, Laura-D., Reczek, Elizabeth-E., Corinna-Cosmas, Demicco, Elizabeth-G., McCurrach, Mila-E., Lowe, Scott-W.,Tyler-Jacks. (2000). PERP, an apoptosis-associated target of p53, is a novel member of the PMP-22/gas3 family. Genes Dev.
Ben-Porath, Kozak, C.A.,Benvenisty, N. (1998). Chromosomal mapping of Tmp (Emp1), Xmp (Emp2), and Ymp (Emp3), genes encoding membrane proteins related to Pmp22. Genomics.
Bianchi, B.R., Lynch, K.J., Touma, E., Niforatos, W., Burgard, E.C.,Alexander, K.M. (1999). Pharmacological characterization of recombinant human and rat P2X receptor subtypes. Eur J Pharmacol.
Bottinger, E.P.,Bitzer, M. (2002). TGF-beta signaling in renal disease. J Am Soc Nephrol.
Brodin, G., Dijke, P., Funa, K., Heldin, C.H.,Landstrom, M. (1999). Increased smad expression and activation are associated with apoptosis in normal and malignant prostate after castration. cancer research.
Burnstock, G. (2013). Purinergic signalling in the lower urinary tract. Acta Physiol (Oxf).
Chen, Yi-Hsien, Wu, Li-Ching, Wu, Wen-Ren, Lin, Hung-Jung, Lee, Sung-Wei, Lin, Ching-Yih, Chang, Shih-Lun, Chow, Nan-Haw, Huang, Hsuan-Ying, Li, Chien-Feng, Hsu, Han-Ping,Shiu, Yow-Ling. (2012). Loss of epithelial membrane protein-2 expression confers an independent prognosticator in nasopharyngeal carcinoma: a cohort study. BMJ Open.
Claassen, G.F.,Hann, S.R. (2000). A role for transcriptional repression of p21CIP1 by c-Myc in overcoming transforming growth factor beta -induced cell-cycle arrest. Proc Natl Acad Sci U S A.
Compérat, E., Varinot, J., Moroch, J., Eymerit-Morin, C.,Brimo, F. (2018). A practical guide to bladder cancer pathology. Nature Reviews Urology.
Derynck, R. (1994). Tgf-Beta-Receptor-Mediated Signaling. Trends Biochem Sciences.
Duckworth, B.C., Weaver, J.S.,Ruderman, J.V. (2002). G2 arrest in Xenopus oocytes depends on phosphorylation of cdc25 by protein kinase A. Proc Natl Acad Sci U S A.
Feng, X.H., Lin, X.,Derynck, R. (2000). Smad2, Smad3 and Smad4 cooperate with Sp1 to induce p15(Ink4B) transcription in response to TGF-beta. The EMBO Journal.
Gorodeski, G.I. (2009). P2X7-mediated chemoprevention of epithelial cancers. Expert Opinion on Therapeutic Targets.
Grande, J.P. (1997). Role of transforming growth factor-beta in tissue injury and repair. Proc Soc Exp Biol Med.
Han, I.,Kudlow, J.E. (1997). Reduced O glycosylation of Sp1 is associated with increased proteasome susceptibility. Mol Cell Biol.
Hocevar, B.A.,Howe, P.H. (1998). Mechanisms of TGF-beta-induced cell cycle arrest. . Miner Electrol Metab.
Jeffrey-Donovan,Joyce-Slingerland. (2000). Transforming growth factor-β and breast cancer: Cell cycle arrest by transforming growth factor-β and its disruption in cancer. Breast Cancer Research.
Knowles, M.A.,Hurst, C.D. . (2015). Molecular biology of bladder cancer: new insights into pathogenesis and clinical diversity. Nat Rev Cancer.
Li-Yang,Moses, Harold-L. (2008). Transforming Growth Factor B: Tumor Suppressor or Promoter?Are Host Immune Cells the Answer? cancer research.
Li, Chien-Feng, Wu, Wen-Jeng, Wu, Wen-Ren, Liao, Yu-Jing, Chen, Lih-Ren, Huang, Chun-Nung, Li, Ching-Chia, Li, Wei-Ming, Huang, Hsuan-Ying, Chen, Yi-Ling, Liang, Shih-Shin, Chow, Nan-Haw,Shiue, Yow-Ling. (2015). The cAMP responsive element binding protein 1 transactivates epithelial membrane protein 2, a potential tumor suppressor in the urinary bladder urothelial carcinoma. Oncotarget.
Long-Zhang, Fangfang-Zhou,Dijke, Peter-ten. (2013). Signaling interplay between transforming growth factor-b receptor and PI3K/AKT pathways in cancer. Trends in Biochemical Sciences.
Mertens-Talcott, S.U., Chintharlapalli, S., Li, X.,Safe, S. (2007). The oncogenic microRNA-27a targets genes that regulate specificity protein transcription factors and the G2-M checkpoint in MDA-MB-231 breast cancer cells. cancer research.
Morgan, David-O. (2007). Cell Cycle: Principles of Control. Yale J Biol Med.
Owen, G.I., Richer, J.K., Tung, L., Takimoto, G.,Horwitz, K.B. (1998). Progesterone regulates transcription of the p21(WAF1) cyclin-dependent kinase inhibitor gene through Sp1 and CBP/p300. The Journal of biological chemistry.
Park-C.Y., Kim-D.K.,Sheen-Y.Y. (2011). EW-7203, a novel small molecule inhibitor of transforming growth factor-β (TGF-β) type I receptor/activin receptor-like kinase-5, blocks TGF-β1-mediated epithelial-to-mesenchymal transition in mammary epithelial cells. Cancer Sciences.
Parker, L.L.,Piwnica-Worms, H. (1992). Inactivation of the p34cdc2-cyclin B complex by the human WEE1 tyrosine kinase. Science.
Rahimi, R.A.,Leof, B.E. (2007). TGF-b Signalling: A Tale of Two Responses. Journal of Cellular Biochemistry.
Surprenant, A., Rassendren, F., Kawashima, E.,North, R.A. (1996). The cytolytic P2Z receptor for extracellular ATP identified as a P2X receptor (P2X7). Science.
Wadehra, M., Forbes, A., Pushkarna, N., Goodglick, L., Gordon, L. K., Williams, C. J.,Braun, J. (2005). Epithelial membrane protein-2 regulates surface expression of alphavbeta3 integrin in the endometrium. Dev Biol.
Wadehra, M., Sulur, G.G., Braun, J., Gordon, L.K.,Goodglick, L. (2003). Epithelial membrane protein-2 is expressed in discrete anatomical regions of the eye. Exp Mol Pathol.
Wilson, H.L., Wilson, S.A., Surprenant, A.,North, R.A. (2002). Epithelial membrane proteins induce membrane blebbing and interact with the P2X7 receptor C terminus. J Biol Chem.
Wong, C.F., Barnes, L.M., Dahler, A.L., Smith, L., Popa, C., Serewko-Auret, M.M.,Saunders, N.A. (2005). E2F suppression and Sp1 overexpression are sufficient to induce the differentiation-specific marker, transglutaminase type 1, in a squamous cell carcinoma cell line. oncogene.
Wu, J., Xue, L., Weng, M., Sun, Y., Zhang, Z., Wang, W.,Tong, T. (2007). Sp1 is essential for p16 expression in human diploid fibroblasts during senescence. PLoS ONE.
Yabing-Mu, Gudey, Shyam-Kumar,Maréne-Landström. (2012). Non-Smad signaling pathways. Cell and Tissue Research.
Zhang, Ying-E. (2009). Non-Smad pathways in TGF-β signaling. Cell Research.
Zhou, G.S., Lee, S.C., Yao, Z.B.,Tan, T.H. (1999). Hematopoietic progenitor kinase 1 is a component of transforming growth factor beta-induced c-Jun N-terminal kinase signaling cascade. Journal of Biological Chemistry.
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