請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77665完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳彥榮(Edward Chern) | |
| dc.contributor.author | Tsu-Ning Lu | en |
| dc.contributor.author | 呂祖寧 | zh_TW |
| dc.date.accessioned | 2021-07-10T22:14:35Z | - |
| dc.date.available | 2021-07-10T22:14:35Z | - |
| dc.date.copyright | 2018-03-01 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-02-21 | |
| dc.identifier.citation | Allemand, E., Hastings, M.L., Murray, M.V., Myers, M.P., and Krainer, A.R. (2007). Alternative splicing regulation by interaction of phosphatase PP2Cγ with nucleic acid–binding protein YB-1. Nature structural & molecular biology 14, 630-638.
Bargou, R.C., Jürchott, K., Wagener, C., Bergmann, S., Metzner, S., Bommert, K., Mapara, M.Y., Winzer, K.-J., Dietel, M., and Dörken, B. (1997). Nuclear localization and increased levels of transcription factor YB-1 in primary human breast cancers are associated with intrinsic MDR1 gene expression. Nature medicine 3, 447-450. Bonnet, D., and Dick, J.E. (1997). Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nature medicine 3, 730-737. Brabletz, T., Jung, A., Spaderna, S., Hlubek, F., and Kirchner, T. (2005). Opinion: migrating cancer stem cells - an integrated concept of malignant tumour progression. Nature reviews Cancer 5, 744-749. Chaffer, C.L., and Weinberg, R.A. (2011). A perspective on cancer cell metastasis. Science 331, 1559-1564. Chao, H.M., Huang, H.X., Chang, P.H., Tseng, K.C., Miyajima, A., and Chern, E. (2017). Y-box binding protein-1 promotes hepatocellular carcinoma-initiating cell progression and tumorigenesis via Wnt/beta-catenin pathway. Oncotarget 8, 2604-2616. Cheng, C., and Sharp, P.A. (2006). Regulation of CD44 alternative splicing by SRm160 and its potential role in tumor cell invasion. Molecular and cellular biology 26, 362-370. Clevers, H. (2011). The cancer stem cell: premises, promises and challenges. Nature medicine, 313-319. Cohen, S., Ma, W., Valova, V., Algie, M., Harfoot, R., Woolley, A., Robinson, P., and Braithwaite, A. (2010). Genotoxic stress-induced nuclear localization of oncoprotein YB-1 in the absence of proteolytic processing. Oncogene 29, 403-410. Cooper, T.A. (2005). Use of minigene systems to dissect alternative splicing elements. Methods 37, 331-340. Corbo, C., Orrù, S., and Salvatore, F. (2013). SRp20: an overview of its role in human diseases. Biochemical and biophysical research communications 436, 1-5. Dahl, E., En-Nia, A., Wiesmann, F., Krings, R., Djudjaj, S., Breuer, E., Fuchs, T., Wild, P.J., Hartmann, A., Dunn, S.E., et al. (2009). Nuclear detection of Y-box protein-1 (YB-1) closely associates with progesterone receptor negativity and is a strong adverse survival factor in human breast cancer. BMC Cancer 9, 410. Davies, A.H., Reipas, K.M., Pambid, M.R., Berns, R., Stratford, A.L., Fotovati, A., Firmino, N., Astanehe, A., Hu, K., Maxwell, C., et al. (2014). YB-1 transforms human mammary epithelial cells through chromatin remodeling leading to the development of basal-like breast cancer. Stem cells (Dayton, Ohio) 32, 1437-1450. Didier, D.K., Schiffenbauer, J., Woulfe, S.L., Zacheis, M., and Schwartz, B.D. (1988). Characterization of the cDNA encoding a protein binding to the major histocompatibility complex class II Y box. Proceedings of the National Academy of Sciences 85, 7322-7326. Dolfini, D., and Mantovani, R. (2013). Targeting the Y/CCAAT box in cancer: YB-1 (YBX1) or NF-Y? Cell Death & Differentiation 20, 676-685. Eliseeva, I., Kim, E., Guryanov, S., Ovchinnikov, L., and Lyabin, D. (2011). Y-box-binding protein 1 (YB-1) and its functions. Biochemistry (Moscow) 76, 1402-1433. Evdokimova, V., Tognon, C., Ng, T., Ruzanov, P., Melnyk, N., Fink, D., Sorokin, A., Ovchinnikov, L.P., Davicioni, E., and Triche, T.J. (2009). Translational activation of snail1 and other developmentally regulated transcription factors by YB-1 promotes an epithelial-mesenchymal transition. Cancer cell 15, 402-415. Fu, Y., Huang, B., Shi, Z., Han, J., Wang, Y., Huangfu, J., and Wu, W. (2013). SRSF1 and SRSF9 RNA binding proteins promote Wnt signalling-mediated tumorigenesis by enhancing beta-catenin biosynthesis. EMBO molecular medicine 5, 737-750. Fujii, T., Kawahara, A., Basaki, Y., Hattori, S., Nakashima, K., Nakano, K., Shirouzu, K., Kohno, K., Yanagawa, T., Yamana, H., et al. (2008). Expression of HER2 and estrogen receptor alpha depends upon nuclear localization of Y-box binding protein-1 in human breast cancers. Cancer research 68, 1504-1512. Fujii, T., Seki, N., Namoto-Matsubayashi, R., Takahashi, H., Inoue, Y., Toh, U., Kage, M., and Shirouzu, K. (2009). YB-1 prevents apoptosis via the mTOR/STAT3 pathway in HER-2-overexpressing breast cancer cells. Galiana-Arnoux, D., Lejeune, F., Gesnel, M.C., Stevenin, J., Breathnach, R., and Del Gatto-Konczak, F. (2003). The CD44 alternative v9 exon contains a splicing enhancer responsive to the SR proteins 9G8, ASF/SF2, and SRp20. In The Journal of biological chemistry, pp. 32943-32953. Giménez‐Bonafé, P., Fedoruk, M.N., Whitmore, T.G., Akbari, M., Ralph, J.L., Ettinger, S., Gleave, M.E., and Nelson, C.C. (2004). YB‐1 is upregulated during prostate cancer tumor progression and increases P‐glycoprotein activity. The Prostate 59, 337-349. Goncalves, V., Matos, P., and Jordan, P. (2008). The beta-catenin/TCF4 pathway modifies alternative splicing through modulation of SRp20 expression. RNA (New York, NY) 14, 2538-2549. Ha, B., Lee, E.B., Cui, J., Kim, Y., and Jang, H.H. (2015). YB-1 overexpression promotes a TGF-β1-induced epithelial–mesenchymal transition via Akt activation. Biochemical and biophysical research communications 458, 347-351. Haggar, F.A., and Boushey, R.P. (2009). Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clinics in colon and rectal surgery 22, 191-197. Hanahan, D., and Weinberg, R.A. (2011). Hallmarks of cancer: the next generation. cell 144, 646-674. Heider, K.H., Kuthan, H., Stehle, G., and Munzert, G. (2004). CD44v6: a target for antibody-based cancer therapy. Cancer immunology, immunotherapy : CII 53, 567-579. Huang, C.S., Shen, C.Y., Wang, H.W., Wu, P.E., and Cheng, C.W. (2007). Increased expression of SRp40 affecting CD44 splicing is associated with the clinical outcome of lymph node metastasis in human breast cancer. Clinica chimica acta; international journal of clinical chemistry 384, 69-74. Jemal, A., Bray, F., Center, M.M., Ferlay, J., Ward, E., and Forman, D. (2011). Global cancer statistics. CA: a cancer journal for clinicians 61, 69-90. Jia, R., Li, C., McCoy, J.P., Deng, C.X., and Zheng, Z.M. (2010). SRp20 is a proto-oncogene critical for cell proliferation and tumor induction and maintenance. International journal of biological sciences 6, 806-826. Karni, R., de Stanchina, E., Lowe, S.W., Sinha, R., Mu, D., and Krainer, A.R. (2007). The gene encoding the splicing factor SF2/ASF is a proto-oncogene. Nature structural & molecular biology 14, 185-193. Kosnopfel, C., Sinnberg, T., and Schittek, B. (2014). Y-box binding protein 1–a prognostic marker and target in tumour therapy. European journal of cell biology 93, 61-70. Lasham, A., Samuel, W., Cao, H., Patel, R., Mehta, R., Stern, J.L., Reid, G., Woolley, A.G., Miller, L.D., and Black, M.A. (2011). YB-1, the E2F pathway, and regulation of tumor cell growth. Journal of the National Cancer Institute 104, 133-146. Lee, T.K.W., Castilho, A., Cheung, V.C.H., Tang, K.H., Ma, S., and Ng, I.O.L. (2011). CD24+ liver tumor-initiating cells drive self-renewal and tumor initiation through STAT3-mediated NANOG regulation. Cell stem cell 9, 50-63. Li, C., Heidt, D.G., Dalerba, P., Burant, C.F., Zhang, L., Adsay, V., Wicha, M., Clarke, M.F., and Simeone, D.M. (2007). Identification of pancreatic cancer stem cells. Cancer research 67, 1030-1037. Loh, T.J., Moon, H., Cho, S., Jung, D.W., Hong, S.E., Kim, D.H., Green, M.R., Zheng, X., Zhou, J., and Shen, H. (2014). SC35 promotes splicing of the C5-V6-C6 isoform of CD44 pre-mRNA. Oncology reports 31, 273-279. Matter, N., Herrlich, P., and Konig, H. (2002). Signal-dependent regulation of splicing via phosphorylation of Sam68. Nature 420, 691-695. Maurya, P.K., Mishra, A., Yadav, B.S., Singh, S., Kumar, P., Chaudhary, A., Srivastava, S., Murugesan, S.N., and Mani, A. (2017). Role of Y Box Protein-1 in cancer: As potential biomarker and novel therapeutic target. Journal of Cancer 8, 1900-1907. Naro, C., and Sette, C. (2013). Phosphorylation-mediated regulation of alternative splicing in cancer. International journal of cell biology 2013, 151839. Nishisho, I., Nakamura, Y., Miyoshi, Y., Miki, Y., Ando, H., Horii, A., Koyama, K., Utsunomiya, J., Baba, S., and Hedge, P. (1991). Mutations of chromosome 5q21 genes in FAP and colorectal cancer patients. Science 253, 665-669. O'Brien, C.A., Pollett, A., Gallinger, S., and Dick, J.E. (2007). A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445, 106-110. Orian-Rousseau, V. (2010). CD44, a therapeutic target for metastasising tumours. European journal of cancer (Oxford, England : 1990) 46, 1271-1277. Orian-Rousseau, V. (2015). CD44 Acts as a Signaling Platform Controlling Tumor Progression and Metastasis. Frontiers in immunology 6, 154. Pang, R., Law, W.L., Chu, A.C., Poon, J.T., Lam, C.S., Chow, A.K., Ng, L., Cheung, L.W., Lan, X.R., Lan, H.Y., et al. (2010). A subpopulation of CD26+ cancer stem cells with metastatic capacity in human colorectal cancer. Cell stem cell 6, 603-615. Prochazka, L., Tesarik, R., and Turanek, J. (2014). Regulation of alternative splicing of CD44 in cancer. Cell Signal 26, 2234-2239. Raffetseder, U., Frye, B., Rauen, T., Jurchott, K., Royer, H.D., Jansen, P.L., and Mertens, P.R. (2003). Splicing factor SRp30c interaction with Y-box protein-1 confers nuclear YB-1 shuttling and alternative splice site selection. The Journal of biological chemistry 278, 18241-18248. Rangaswami, H., Bulbule, A., and Kundu, G.C. (2006). Osteopontin: role in cell signaling and cancer progression. Trends in cell biology 16, 79-87. Reya, T., Morrison, S.J., Clarke, M.F., and Weissman, I.L. (2001). Stem cells, cancer, and cancer stem cells. Nature 414, 105-111. Senbanjo, L.T., and Chellaiah, M.A. (2017). CD44: A Multifunctional Cell Surface Adhesion Receptor Is a Regulator of Progression and Metastasis of Cancer Cells. Frontiers in cell and developmental biology 5. Shibahara, K., Sugio, K., Osaki, T., Uchiumi, T., Maehara, Y., Kohno, K., Yasumoto, K., Sugimachi, K., and Kuwano, M. (2001). Nuclear expression of the Y-box binding protein, YB-1, as a novel marker of disease progression in non-small cell lung cancer. Clinical cancer research 7, 3151-3155. Shiota, M., Izumi, H., Onitsuka, T., Miyamoto, N., Kashiwagi, E., Kidani, A., Yokomizo, A., Naito, S., and Kohno, K. (2008). Twist promotes tumor cell growth through YB-1 expression. Cancer research 68, 98-105. Singh, G., and Cooper, T.A. (2006). Minigene reporter for identification and analysis of cis elements and trans factors affecting pre-mRNA splicing. Biotechniques 41, 177. Stickeler, E., Fraser, S.D., Honig, A., Chen, A.L., Berget, S.M., and Cooper, T.A. (2001). The RNA binding protein YB-1 binds A/C-rich exon enhancers and stimulates splicing of the CD44 alternative exon v4. The EMBO journal 20, 3821-3830. Teschendorf, C., Warrington, K.H., Jr., Siemann, D.W., and Muzyczka, N. (2002). Comparison of the EF-1 alpha and the CMV promoter for engineering stable tumor cell lines using recombinant adeno-associated virus. Anticancer research 22, 3325-3330. To, K., Fotovati, A., Reipas, K.M., Law, J.H., Hu, K., Wang, J., Astanehe, A., Davies, A.H., Lee, L., and Stratford, A.L. (2010). Y-box binding protein-1 induces the expression of CD44 and CD49f leading to enhanced self-renewal, mammosphere growth, and drug resistance. Cancer research 70, 2840-2851. Todaro, M., Gaggianesi, M., Catalano, V., Benfante, A., Iovino, F., Biffoni, M., Apuzzo, T., Sperduti, I., Volpe, S., Cocorullo, G., et al. (2014). CD44v6 is a marker of constitutive and reprogrammed cancer stem cells driving colon cancer metastasis. Cell stem cell 14, 342-356. Twyffels, L., Gueydan, C., and Kruys, V. (2011). Shuttling SR proteins: more than splicing factors. The FEBS journal 278, 3246-3255. Valent, P., Bonnet, D., De Maria, R., Lapidot, T., Copland, M., Melo, J.V., Chomienne, C., Ishikawa, F., Schuringa, J.J., Stassi, G., et al. (2012). Cancer stem cell definitions and terminology: the devil is in the details. Nature reviews Cancer 12, 767-775. Wang, J., Xiao, L., Luo, C.H., Zhou, H., Zeng, L., Zhong, J., Tang, Y., Zhao, X.H., Zhao, M., and Zhang, Y. (2015). CD44v6 promotes beta-catenin and TGF-beta expression, inducing aggression in ovarian cancer cells. Molecular medicine reports 11, 3505-3510. Wang, Y.W., Chen, H.H., Wu, M.S., and Chiu, H.M. (2017). Current status and future challenge of population-based organized colorectal cancer screening: Lesson from the first decade of Taiwanese program. Journal of the Formosan Medical Association = Taiwan yi zhi. Wei, W.J., Mu, S.R., Heiner, M., Fu, X., Cao, L.J., Gong, X.F., Bindereif, A., and Hui, J. (2012). YB-1 binds to CAUC motifs and stimulates exon inclusion by enhancing the recruitment of U2AF to weak polypyrimidine tracts. Nucleic acids research 40, 8622-8636. Yan, X.B., Zhu, Q.C., Chen, H.Q., Peng, J.Y., Chao, H.L., Du, H.X., Wang, Z.G., and Jin, Z.M. (2014). Knockdown of Yboxbinding protein1 inhibits the malignant progression of HT29 colorectal adenocarcinoma cells by reversing epithelialmesenchymal transition. Molecular medicine reports 10, 2720-2728. Yasen, M., Kajino, K., Kano, S., Tobita, H., Yamamoto, J., Uchiumi, T., Kon, S., Maeda, M., Obulhasim, G., and Arii, S. (2005). The up-regulation of Y-box binding proteins (DNA binding protein A and Y-box binding protein-1) as prognostic markers of hepatocellular carcinoma. Clinical cancer research 11, 7354-7361. Yoshino, H., Enokida, H., Chiyomaru, T., Tatarano, S., Hidaka, H., Yamasaki, T., Gotannda, T., Tachiwada, T., Nohata, N., Yamane, T., et al. (2012). Tumor suppressive microRNA-1 mediated novel apoptosis pathways through direct inhibition of splicing factor serine/arginine-rich 9 (SRSF9/SRp30c) in bladder cancer. Biochemical and biophysical research communications 417, 588-593. Zeilstra, J., Joosten, S.P., van Andel, H., Tolg, C., Berns, A., Snoek, M., van de Wetering, M., Spaargaren, M., Clevers, H., and Pals, S.T. (2014). Stem cell CD44v isoforms promote intestinal cancer formation in Apc(min) mice downstream of Wnt signaling. Oncogene 33, 665-670. Zhang, P., Fu, C., Bai, H., Song, E., and Song, Y. (2014). CD44 variant, but not standard CD44 isoforms, mediate disassembly of endothelial VE‐cadherin junction on metastatic melanoma cells. FEBS letters 588, 4573-4582. Zhu, S., Chen, Z., Katsha, A., Hong, J., Belkhiri, A., and El-Rifai, W. (2016). Regulation of CD44E by DARPP-32-dependent activation of SRp20 splicing factor in gastric tumorigenesis. Oncogene 35, 1847-1856. Zoller, M. (2011). CD44: can a cancer-initiating cell profit from an abundantly expressed molecule? Nature reviews Cancer 11, 254-267. Zuo, P., and Manley, J.L. (1993). Functional domains of the human splicing factor ASF/SF2. The EMBO journal 12, 4727. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77665 | - |
| dc.description.abstract | 癌症的復發及轉移，被認為是造成大腸癌病患低生存率的原因。近年來許多研究發現，癌幹細胞具有自我更新、抗藥的能力，且在化學治療的過程中，能夠生存下來並轉移。癌幹細胞的辨識多半是藉由其表面的幹細胞標記。其中，CD44v6 就是一個極具代表性的大腸癌幹細胞標記，並且被證明會促進大腸癌的轉移。另外，臨床數據也指出，低表現量的 CD44v6 提升了大腸癌病患的生存率。然而，大腸癌幹細胞為什麼傾向將 CD44 的 mRNA 剪接成 CD44v6 這個形式仍然未知。在本篇論文中，發現兩個與剪接反應有關的因子在大腸癌腫瘤組織中有很高的表現量，且能夠促進 CD44v6 轉錄層次的表現量。另外，本研究也證明，兩因子間有交互作用，且影響兩因子入核的現象。然而，在之後進行 minigene reporter assay，卻發現 兩因子似乎無法藉由調控選擇性剪接反應的機制，使外顯子v6被保留下來。因此本論文推測其可能藉由別的機制，例如：調控轉錄，達到調節 CD44v6 表現量的目的。綜上所述，此篇研究說明了關鍵二因子調控了 CD44v6 的表現量，以及細胞轉移的能力，在大腸癌幹細胞中扮演了很重要的角色。 | zh_TW |
| dc.description.abstract | In colorectal cancer (CRC), relapse and metastasis are the main reasons that decrease patients’ probability of survival. In recent years, many studies revealed that the existence of cancer stem cell (CSC) resulted in difficulty in completely curing cancer. The CSC which is defined by its capacity for self-renewal and drug resistance is believed to survive after cancer therapies and then starts metastasis. In tumor tissue, CSCs are commonly identified by their cell surface markers. According to previous studies, CD44v6 was regarded as a crucial cell-surface marker of CSC that confers the ability to promote colorectal cancer metastasis. Besides, lower CD44v6 expression increases CRC patients’ survival rate in clinical studies. However, the alternative splicing mechanism of CD44v6 in CSC is still unknown. In this thesis, I reported that Factor A and Factor B highly expressed in primary colon cancer tissues comparing to their adjacent normal colon tissues and were positively correlated with CD44v6 expression. Moreover, I clarified that there was an interaction between Factor A and Factor B by co-immunoprecipitation. Besides, nucleus-translocated Factor B resulted in poor prognosis in cancer progression. I also found Factor A had ability to enhance the nucleic subcellular localization of Factor B. However, while utilizing minigene reporter assay, I recognized that Factor A and Factor B seemed not regulate CD44v6 expression by changing alternative splicing site choice. Therefore, I supposed that Factor A and Factor B might regulate the expression of CD44v6 via other mechanism such as transcriptional regulation. Overall, these finding revealed that Factor A and Factor B played an important role in regulating CD44v6 expression and cell migration ability which were both considered to be major features of colon CSC. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-10T22:14:35Z (GMT). No. of bitstreams: 1 ntu-107-R04B22070-1.pdf: 44862162 bytes, checksum: 885ffeda59f031789047ed5f01b2a8a3 (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | Table of Contents
⼝試委員會審定書 --------------------------------------------------------------------------------------i 中⽂摘要 --------------------------------------------------------------------------------------------------ii Abstract --------------------------------------------------------------------------------------------iii Table of Contents ----------------------------------------------------------------------------------v Chapter 1. Introduction --------------------------------------------------------------------------1 1.1. Colorectal cancer ---------------------------------------------------------------------------1 1.2. Cancer stem cell ---------------------------------------------------------------------------1 1.3. CD44 ----------------------------------------------------------------------------------------3 1.3.1. Introduction of CD44 ----------------------------------------------------------------3 1.3.2. CD44v6 regarded as a cancer stem cell marker ----------------------------------4 1.3.3. Regulation of CD44 alternative splicing in cancer ------------------------------5 1.4. Serine/arginine-rich splicing factor (SRSF) --------------------------------------------6 1.4.1. Introduction of SRSF ----------------------------------------------------------------6 1.4.2. CD44 alternative splicing affected by SRSFs ------------------------------------7 1.4.3. SRSF9 regarded as an oncogene ------------------------------------------------------8 1.4.4. SRSF9 interacted with Y box binding protein 1 (YB1) managing alternative splicing site selection -----------------------------------------------------------------------9 1.5. Y box binding protein 1 (YB1)- ----------------------------------------------------------9 1.5.1. Introduction of Y box binding protein 1 ------------------------------------------9 1.5.2. YB-1 was regarded as an oncoprotein -------------------------------------------10 1.5.3. YB-1 affected CD44 alternative splicing ----------------------------------------11 Chapter 2. Motivation and Aim ---------------------------------------------------------------13 Chapter 3. Materials and Methods -----------------------------------------------------------15 3.1. Cell culture --------------------------------------------------------------------------------15 3.2. Minigene Reporter assay ----------------------------------------------------------------15 3.2.1. Construction of CD44v6 minigene plasmid ------------------------------------15 3.2.2. Cell culture and Transfection -----------------------------------------------------16 3.2.3. RNA Extraction and RT-PCR -----------------------------------------------------16 3.3. Lentivirus package vector ---------------------------------------------------------------17 3.3.1. pCDH system -----------------------------------------------------------------------17 3.3.2. pLKO_TRC005 system ------------------------------------------------------------17 3.3.3. Virus production --------------------------------------------------------------------18 3.3.4. Virus infection ----------------------------------------------------------------------19 3.4. Cellular RNA extraction and reverse transcription ----------------------------------19 3.5. Quantitative real-time polymerase chain reaction (qRT-PCR) ---------------------20 3.6. Cellular protein extraction --------------------------------------------------------------20 3.7. SDS-PAGE and Western blotting ------------------------------------------------------21 3.8. Co-immunoprecipitation (Co-IP) ------------------------------------------------------22 3.9. Transwell migration assay ---------------------------------------------------------------23 3.10. Cell immunofluorescence staining ---------------------------------------------------24 3.11. Image processing ------------------------------------------------------------------------25 3.12. Statistical analysis ----------------------------------------------------------------------25 Chapter 4. Results -------------------------------------------------------------------------------26 4.1. The expression of CD44v6, SRSF9 and YB1 was positively correlated with cancer progression -----------------------------------------------------------------------------26 4.2. SRSF9 enhanced cell migration ability ------------------------------------------------27 4.3. YB1 enhanced cell migration ability- --------------------------------------------------27 4.4. SRSF9 and YB1 affected CD44v6 expression not through changing the alternative splicing site choice ---------------------------------------------------------------29 4.5. Nuclear shuttling of endogenous YB1 was detected in DLD-1 overexpressed SRSF9 -------------------------------------------------------------------------------------------30 Chapter 5. Conclusion and Discussion ------------------------------------------------------32 Chapter 6. Figures and Tables ----------------------------------------------------------------38 Table 1. List of primers used in minigene reporter assay --------------------------------38 Table 2. Thermal profile of RT-PCR --------------------------------------------------------38 Table 3. List of pRTPCR primers- -----------------------------------------------------------39 Table 4. List of antibodies --------------------------------------------------------------------40 1. CD44 and CD44v6 highly expressed in primary colon cancer tissues -------------42 Figure 2. SRSF9 and YB1 highly expressed in primary colon cancer tissues ---------44 Figure 3. CD44v6 expression and migration ability regulated by SRSF9 ------------47 Figure 4. YB1 overexpression resulting in increased migration ability but not CD44v6 expression ----------------------------------------------------------------------------50 Figure 5. CD44v6 expression and migration ability repressed by YB1 knockdown -52 Figure 6. SRSF9 and YB1 inducing CD44 variant exon v6 exclusion during alternative splicing ----------------------------------------------------------------------------55 Figure 7. YB1 interacted with non-phosphorylated SRSF9 in DLD-1 YB1 OE ------57 Figure 8. Endogenous YB1 translocated to nucleus during SRSF9 overexpression in DLD-1 -------------------------------------------------------------------------------------------60 Chapter 7. Appendixes --------------------------------------------------------------------------61 Appendix 1. CD44 pre-mRNA and its protein structure ---------------------------------61 Appendix 2. The expressions of CD44s and CD44E in colon cancer -----------------62 Appendix 3. SRSF9 was up-regulated in DLD-1 side population ----------------------63 Chapter 8. References ---------------------------------------------------------------------------64 | |
| dc.language.iso | en | |
| dc.subject | CD44v6 | zh_TW |
| dc.subject | 大腸癌幹細胞 | zh_TW |
| dc.subject | 轉移 | zh_TW |
| dc.subject | 選擇性剪接反應 | zh_TW |
| dc.subject | CD44v6 | en |
| dc.subject | colon cancer stem cells | en |
| dc.subject | alternative splicing | en |
| dc.subject | metastasis | en |
| dc.title | 探討⼤腸癌細胞中調控 CD44v6 表現之機制 | zh_TW |
| dc.title | Regulation of CD44v6 Expression in Colon Cancer Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 廖憶純,黃楓婷 | |
| dc.subject.keyword | 大腸癌幹細胞,轉移,CD44v6,選擇性剪接反應, | zh_TW |
| dc.subject.keyword | colon cancer stem cells,metastasis,CD44v6,alternative splicing, | en |
| dc.relation.page | 71 | |
| dc.identifier.doi | 10.6342/NTU201800547 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2018-02-21 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科技學系 | zh_TW |
| 顯示於系所單位： | 生化科技學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-107-R04B22070-1.pdf 未授權公開取用 | 43.81 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。