簡易檢索 / 詳目顯示

研究生: 林新智
Lin, Shin-Chin
論文名稱: 探討在前列腺癌藥物安可坦抗藥性中NUDT21蛋白質後修飾所扮演的角色
To investigate the role of NUDT21 protein post-translational modification (PTM) in enzalutamide resistance
指導教授: 林世杰
Lin, Shih-Chieh
學位類別: 碩士
Master
系所名稱: 醫學院 - 生理學研究所
Department of Physiology
論文出版年: 2021
畢業學年度: 109
語文別: 英文
論文頁數: 60
中文關鍵詞: 安可坦抗藥性前列腺癌Nudix水解酶21替代性聚腺苷酸化甲基化
外文關鍵詞: prostate cancer, enzalutamide resistance, NUDT21, alternative polyadenylation, methylation
相關次數: 點閱:102下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • Abstract I 中文摘要 III 致謝 V Content VII List of figures IX List of tables IX Introduction 1 Materials and methods 7 Cell culture 7 Site-directed mutatgenesis 7 siRNA transfection 8 Primer design 9 Plasmid extraction 9 Isolation of RNA and Protein 10 Western Blot 11 Immunoprecipitation (IP) 12 RNA-Immunoprecipitation (RIP) 13 Chromatin-Immunoprecipitation (ChIP) 14 Immunofluorescence (IF) 17 Quantitative real-time reverse transcription PCR (qRT-PCR) 18 3'PCR 19 MTS assay 21 The Soft Agar Colony Formation Assay 21 Quantification and Statistical Analysis 22 Results 22 NUDT21 was mono-methylated at R15 22 NUDT21 R15me1 modification was mediated by PRMT7 24 Androgen-induced NUDT21 R15me1 medication was mediated through upregulation of PRMT7 expression via androgen receptor (AR) dependent manner in LNCaP cells 25 Inhibition of PRMT7 function decreased the sensitivity of enzalutamide in LNCaP cells 26 NUDT21 R15me1 will change the choice of the polyadenylation site (PAS) in vitro 27 R15me1 modification of NUDT21 promoted its protein degradation 28 The expression of NUDT21 R15me1 was decreased while its total form was increased in prostate cancer tissue 29 Discussion 29 References 32 Appendix 54 < Reagents and buffer used in cell cultured > 54 < Reagents and buffer used in Western blotting > 55 < Reagents used for RT-PCR and qRT-PCR> 57 < Reagents used for Point mutation> 58 < Reagents used for Plasmid extration> 58

    [1] M. Campbell et al., "Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen," (in eng), The Journal of biological chemistry, vol. 287, no. 8, pp. 5806-5818, 2012.
    [2] F. Herrmann, P. Pably, C. Eckerich, M. T. Bedford, and F. O. Fackelmayer, "Human protein arginine methyltransferases in vivo--distinct properties of eight canonical members of the PRMT family," (in eng), J. Cell Sci., vol. 122, no. Pt 5, pp. 667-77, Mar 1 2009.
    [3] T. R. Ferreira et al., "PRMT7 regulates RNA-binding capacity and protein stability in Leishmania parasites," (in eng), Nucleic Acids Res., vol. 48, no. 10, pp. 5511-5526, Jun 4 2020.
    [4] Y.-C. Wang, S. E. Peterson, and J. F. Loring, "Protein post-translational modifications and regulation of pluripotency in human stem cells," Cell Res., vol. 24, no. 2, pp. 143-160, 2014.
    [5] J. M. Hsu, C. W. Li, Y. J. Lai, and M. C. Hung, "Posttranslational Modifications of PD-L1 and Their Applications in Cancer Therapy," (in eng), Cancer Res., vol. 78, no. 22, pp. 6349-6353, Nov 15 2018.
    [6] V. Singh, M. Ram, R. Kumar, R. Prasad, B. K. Roy, and K. K. Singh, "Phosphorylation: Implications in Cancer," (in eng), Protein J., vol. 36, no. 1, pp. 1-6, Feb 2017.
    [7] M. Audagnotto and M. Dal Peraro, "Protein post-translational modifications: In silico prediction tools and molecular modeling," Computational and structural biotechnology journal, vol. 15, pp. 307-319, 2017.
    [8] R. S. Blanc and S. Richard, "Arginine Methylation: The Coming of Age," (in eng), Mol. Cell, vol. 65, no. 1, pp. 8-24, Jan 5 2017.
    [9] J. Zhang, L. Jing, M. Li, L. He, and Z. Guo, "Regulation of histone arginine methylation/demethylation by methylase and demethylase," Mol. Med. Report., vol. 19, no. 5, pp. 3963-3971, 2019.
    [10] J. Liu et al., "Arginine methylation‐dependent LSD1 stability promotes invasion and metastasis of breast cancer," EMBO reports, vol. 21, no. 2, p. e48597, 2020.
    [11] Y. Takahashi, H. Daitoku, A. Yokoyama, K. Nakayama, J. D. Kim, and A. Fukamizu, "The C. elegans PRMT-3 possesses a type III protein arginine methyltransferase activity," (in eng), J. Recept. Signal Transduct. Res., vol. 31, no. 2, pp. 168-72, Apr 2011.
    [12] M. M. Szewczyk et al., "Pharmacological inhibition of PRMT7 links arginine monomethylation to the cellular stress response," (in eng), Nat Commun, vol. 11, no. 1, p. 2396, May 14 2020.
    [13] L. Fang et al., "SET1A-Mediated Mono-Methylation at K342 Regulates YAP Activation by Blocking Its Nuclear Export and Promotes Tumorigenesis," (in eng), Cancer Cell, vol. 34, no. 1, pp. 103-118.e9, Jul 9 2018.
    [14] R. M. Baldwin, A. Morettin, and J. Côté, "Role of PRMTs in cancer: Could minor isoforms be leaving a mark?," World J. Biol. Chem., vol. 5, no. 2, p. 115, 2014.
    [15] K. Jain and S. G. Clarke, "PRMT7 as a unique member of the protein arginine methyltransferase family: A review," (in eng), Arch. Biochem. Biophys., vol. 665, pp. 36-45, Apr 15 2019.
    [16] J. Brumbaugh et al., "Nudt21 Controls Cell Fate by Connecting Alternative Polyadenylation to Chromatin Signaling," (in eng), Cell, vol. 172, no. 1-2, pp. 106-120.e21, Jan 11 2018.
    [17] J. Guhaniyogi and G. Brewer, "Regulation of mRNA stability in mammalian cells," (in eng), Gene, vol. 265, no. 1-2, pp. 11-23, Mar 7 2001.
    [18] D. Liu et al., "Systematic variation in mRNA 3'-processing signals during mouse spermatogenesis," (in eng), Nucleic Acids Res., vol. 35, no. 1, pp. 234-46, 2007.
    [19] S. Bienroth, W. Keller, and E. Wahle, "Assembly of a processive messenger RNA polyadenylation complex," (in eng), EMBO J., vol. 12, no. 2, pp. 585-94, Feb 1993.
    [20] B. Tian and J. L. Manley, "Alternative polyadenylation of mRNA precursors," (in eng), Nat. Rev. Mol. Cell Biol., vol. 18, no. 1, pp. 18-30, Jan 2017.
    [21] D. C. Di Giammartino, K. Nishida, and J. L. Manley, "Mechanisms and consequences of alternative polyadenylation," (in eng), Mol. Cell, vol. 43, no. 6, pp. 853-66, Sep 16 2011.
    [22] M. Hoque et al., "Analysis of alternative cleavage and polyadenylation by 3' region extraction and deep sequencing," (in eng), Nat. Methods, vol. 10, no. 2, pp. 133-9, Feb 2013.
    [23] R. Sandberg, J. R. Neilson, A. Sarma, P. A. Sharp, and C. B. Burge, "Proliferating cells express mRNAs with shortened 3' untranslated regions and fewer microRNA target sites," (in eng), Science, vol. 320, no. 5883, pp. 1643-7, Jun 20 2008.
    [24] S. Tan et al., "NUDT21 negatively regulates PSMB2 and CXXC5 by alternative polyadenylation and contributes to hepatocellular carcinoma suppression," (in eng), Oncogene, vol. 37, no. 35, pp. 4887-4900, Aug 2018.
    [25] M. Xiong et al., "NUDT21 inhibits bladder cancer progression through ANXA2 and LIMK2 by alternative polyadenylation," (in eng), Theranostics, vol. 9, no. 24, pp. 7156-7167, 2019.
    [26] "Prostate cancer," (in eng), Nurs. Stand., vol. 30, no. 40, p. 17, Jun 1 2016.
    [27] J. Ferlay et al., "Cancer statistics for the year 2020: An overview," Int. J. Cancer, 2021.
    [28] J. Ferlay et al., "Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012," (in eng), Int. J. Cancer, vol. 136, no. 5, pp. E359-86, Mar 1 2015.
    [29] E. A. Klein, J. Ciezki, P. A. Kupelian, and A. Mahadevan, "Outcomes for intermediate risk prostate cancer: are there advantages for surgery, external radiation, or brachytherapy?," (in eng), Urol. Oncol., vol. 27, no. 1, pp. 67-71, Jan-Feb 2009.
    [30] M. Kirby, C. Hirst, and E. D. Crawford, "Characterising the castration-resistant prostate cancer population: a systematic review," (in eng), Int. J. Clin. Pract., vol. 65, no. 11, pp. 1180-92, Nov 2011.
    [31] D. Robinson et al., "Integrative clinical genomics of advanced prostate cancer," (in eng), Cell, vol. 161, no. 5, pp. 1215-1228, May 21 2015.
    [32] H. I. Scher et al., "Antitumour activity of MDV3100 in castration-resistant prostate cancer: a phase 1-2 study," (in eng), Lancet, vol. 375, no. 9724, pp. 1437-46, Apr 24 2010.
    [33] S. M. Blazie, H. C. Geissel, H. Wilky, R. Joshi, J. Newbern, and M. Mangone, "Alternative Polyadenylation Directs Tissue-Specific miRNA Targeting in Caenorhabditis elegans Somatic Tissues," (in eng), Genetics, vol. 206, no. 2, pp. 757-774, Jun 2017.
    [34] A. Guo et al., "Immunoaffinity enrichment and mass spectrometry analysis of protein methylation," Mol. Cell. Proteomics, vol. 13, no. 1, pp. 372-387, 2014.
    [35] H. C. Lee et al., "YAP1 overexpression contributes to the development of enzalutamide resistance by induction of cancer stemness and lipid metabolism in prostate cancer," (in eng), Oncogene, vol. 40, no. 13, pp. 2407-2421, Apr 2021.
    [36] Z. A. Wang, R. Toivanen, S. K. Bergren, P. Chambon, and M. M. Shen, "Luminal cells are favored as the cell of origin for prostate cancer," (in eng), Cell Rep., vol. 8, no. 5, pp. 1339-46, Sep 11 2014.
    [37] A. Valinezhad Orang, R. Safaralizadeh, and M. Kazemzadeh-Bavili, "Mechanisms of miRNA-Mediated Gene Regulation from Common Downregulation to mRNA-Specific Upregulation," (in eng), Int J Genomics, vol. 2014, p. 970607, 2014.
    [38] A. Quattrone and E. Dassi, "The architecture of the human RNA-binding protein regulatory network," IScience, vol. 21, pp. 706-719, 2019.
    [39] C. P. Masamha et al., "CFIm25 links alternative polyadenylation to glioblastoma tumour suppression," Nature, vol. 510, no. 7505, pp. 412-416, 2014.
    [40] J.-C. Lou et al., "Silencing NUDT21 attenuates the mesenchymal identity of glioblastoma cells via the NF-κB pathway," Front. Mol. Neurosci., vol. 10, p. 420, 2017.
    [41] L. Zhang and W. Zhang, "Knockdown of NUDT21 inhibits proliferation and promotes apoptosis of human K562 leukemia cells through ERK pathway," Cancer Manag. Res., vol. 10, p. 4311, 2018.

    無法下載圖示 校內:不公開
    校外:不公開
    電子論文尚未授權公開,紙本請查館藏目錄
    QR CODE