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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃敏銓 | |
dc.contributor.author | Miao-Juei Huang | en |
dc.contributor.author | 黃妙瑞 | zh_TW |
dc.date.accessioned | 2021-06-16T03:47:59Z | - |
dc.date.available | 2015-03-12 | |
dc.date.copyright | 2015-03-12 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-01-28 | |
dc.identifier.citation | 1. Chen, P.J., et al., Issues and controversies of hepatocellular carcinoma-targeted therapy clinical trials in Asia: experts' opinion. Liver Int, 2010. 30(10): p. 1427-38.
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Hua, D., et al., Polypeptide N-acetylgalactosaminyltransferase 2 regulates cellular metastasis-associated behavior in gastric cancer. Int J Mol Med, 2012. 30(6): p. 1267-74. 12. Wu, Y.M., et al., Mucin glycosylating enzyme GALNT2 regulates the malignant character of hepatocellular carcinoma by modifying the EGF receptor. Cancer Res, 2011. 71(23): p. 7270-9. 13. Kitada, S., et al., Polypeptide N-acetylgalactosaminyl transferase 3 independently predicts high-grade tumours and poor prognosis in patients with renal cell carcinomas. Br J Cancer, 2013. 109(2): p. 472-81. 14. Wang, Z.Q., et al., Role of the polypeptide N-acetylgalactosaminyltransferase 3 in ovarian cancer progression: possible implications in abnormal mucin O-glycosylation. Oncotarget, 2014. 5(2): p. 544-60. 15. Wang, R., et al., The mucin-type glycosylating enzyme polypeptide N-acetylgalactosaminyltransferase 14 promotes the migration of ovarian cancer by modifying mucin 13. Oncol Rep, 2013. 30(2): p. 667-76. 16. 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Roberts, Fibroblast growth factor signaling in liver carcinogenesis. Hepatology, 2014. 59(3): p. 1166-73. 22. Yang, H., et al., MicroRNA-140-5p suppresses tumor growth and metastasis by targeting transforming growth factor beta receptor 1 and fibroblast growth factor 9 in hepatocellular carcinoma. Hepatology, 2013. 58(1): p. 205-17. 23. Lau, C.K., et al., An Akt/hypoxia-inducible factor-1alpha/platelet-derived growth factor-BB autocrine loop mediates hypoxia-induced chemoresistance in liver cancer cells and tumorigenic hepatic progenitor cells. Clin Cancer Res, 2009. 15(10): p. 3462-71. 24. Peng, S., et al., Autocrine vascular endothelial growth factor signaling promotes cell proliferation and modulates sorafenib treatment efficacy in hepatocellular carcinoma. Hepatology, 2014. 60(4): p. 1264-77. 25. Fuchs, B.C., et al., Epidermal growth factor receptor inhibition attenuates liver fibrosis and development of hepatocellular carcinoma. Hepatology, 2014. 59(4): p. 1577-90. 26. 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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/55121 | - |
dc.description.abstract | 肝細胞癌在全球癌症死亡率排名第三位。由於肝細胞癌的惡性行為之特性,在臨床診斷及治療上仍然是一大問題。黏液型醣化修飾作用為蛋白質最常見的轉譯後修飾作用。從過去研究指出,不正常的醣化修飾會參與在癌症發展過程中。GALNT1為黏液型醣化作用步驟中的第一步酵素。但是目前對於GALNT1在肝細胞癌所扮演的角色卻仍然非常不清楚。本研究發現GALNT1在肝細胞癌有高度表現,並且高度表現GALNT1的病人其存活率相較低表現之病人差。從細胞實驗我們發現高表現GALNT1的肝癌細胞可以促進肝癌細胞的惡性行為,例如:細胞遷移和侵襲 ; 反之,降低GALNT1表現則會抑制肝細胞惡性行為。此外,抑制GALNT1表現肝癌細胞會抑制EGF所誘發的細胞遷移和侵襲之惡性行為。並且抑制GALNT1表現可以影響EGFR上的醣化修飾,進而抑制EGFR的磷酸化,進而促進EGFR的降解作用。本研究顯示GALNT1在肝細胞癌中表現量增加,透過抑制GALNT1的表現可以抑制由EGFR所誘發肝癌細胞的惡性行為。未來在肝細胞癌治療研究中,GALNT1是一個具有潛力的方向。 | zh_TW |
dc.description.abstract | Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related mortality worldwide. Management of HCC imposes great challenge due to its malignant characteristics and limited therapeutic drug efficacy. O-glycosylation is a common protein modification, however, aberrant O-glycosylation is associated with many cancer malignancies. GALNT1 is a GalNAc-transferase pivotal in the initiation of protein O-glycosylation, yet, the functional roles of GALNT1 in cancer, particularly HCC, are unknown. In this study we found that GALNT1 is frequently up-regulated in HCC; and higher GALNT1 expression is associated with poorer patient survival. Indeed, overexpression of GALNT1 shows enhanced but knockdown suppressed HCC cell migration and invasion. Further investigation found that knockdown of GALNT1 significantly suppressed EGF-induced HCC cell migration and invasion. Mechanistic investigation shows that knockdown of GALNT1 decreased EGF-triggered EGFR activation and enhanced EGFR degradation by altered O-glycans on EGFR. This study demonstrates that GALNT1 is often overexpressed in HCC and inhibiting GALNT1 expression is sufficient to suppress malignant behaviors of HCC cells by decreasing EGFR signaling suggesting that GALNT1 may be a potential target of therapeutic drug development in the management of HCC. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T03:47:59Z (GMT). No. of bitstreams: 1 ntu-104-D00446003-1.pdf: 11238173 bytes, checksum: c94482fb9ab0fbf1859a4d6205cb6da8 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 口試委員會審定書…………………………………………………………...…………i
Acknowledgment…………………………………………………………...……………ii中文摘要…………………………………………………………...…………………...iii Abstract…………………………………………..…………………………………iv Chapter 1. Introduction……..……………………………………………………………1 Chapter 2. Material and methods…….…………………………………………………..4 2.1 Human tissue samples……………………………………………4 2.2 Cell lines and cell culture…………………………………………...4 2.3 Reagents and antibodies……………………………………………….5 2.4 cDNA synthesis and real-time RT-PCR……………………………....6 2.5 Immunohistochemistry………………………………………...………7 2.6 Plasmid construction………………………………………………….7 2.7 Transfection……………………………………………………………8 2.8 In vivo metastasis assay………………………………………………....8 2.9 Transwell migration and matrigel invasion assays…………………....9 2.10 Cell cycle analysis…………………………………………………….9 2.11 Internalization of EGFR and immunofluorescence microscopy……..10 2.12 Immunoprecipitation and Western blotting…………………………..10 2.13 Gene expression profiling and quantitative RT-PCR validation……..11 2.14 Functional enrichment and network analyzes………………………..12 2.15 Statistical analysis……………………………………………………13 Chapter 3. Results………...…………………………………………………………….14 3.1 GALNT1 is frequently up-regulated in HCC and higher GALNT1 expression levels are associated with poorer overall survival………..14 3.2 GALNT1 expression regulates HCC cell malignant behaviors in vitro……………………………………………………….15 3.3 GALNT1 expression regulates HCC cell malignant behaviors in vivo…………………………………………...……….….16 3.4 GALNT1 knockdown inhibits EGF-induced cell migration and invasion………………………………….……….17 3.5 GALNT1 knockdown reduces EGF-induced EGFR activation and facilitates EGFR degradation………………………………………18 3.6 GALNT1 regulates EGFR O-glycosylation……………………….19 3.7 GALNT1 knockdown enhances EGFR co-localization with EEA1 and LAMP1…………………………………………………………………20 3.8 GALNT1 knockdown affects genes expression………………………..21 Chpater 4. Discussion…………………………………………………………………24 References……………….………….……………………………….…………………30 Table of Figures Figure 1. Figure 1. GALNT1 is frequently up-regulated in HCC…………………..34 Figure 2. GALNT1 is commonly up-regulated in HCC patients from NTUH….35 Figure 3. GALNT1 protein expression is often increased in HCC patients from NTUH…………………………………………………….………………….36 Figure 4. Higher GALNT1 expression is correlated with poorer HCC patient survival………………………………………………………………………37 Figure 5. GALNT1 is differentially expressed in HCC cells………………………...38 Figure 6. GALNT1 knockdown does not affect HCC cell cycle ……………………39 Figure 7. GALNT1 overexpression enhances HCC cell migration and invasion ……..40 Figure 8. GALNT1 knockdown suppresses HCC cell migration and invasion………..41 Figure 9. Knockdown of GALNT1 suppresses HCC cell metastasis in vivo………42 Figure 10. Knockdown of GALNT1 suppresses EGF-induced cell migration…43 Figure 11. Knockdown of GALNT1 suppresses EGF-induced cell invasion…44 Figure 12. Knockdown of GALNT1 reduces EGF-induced EGFR activation and enhances EGFR degradation……………………………………………...…45 Figure 13. EGFR expression levels are moderately correlated with GALNT1 expression levels in HCC tumors…………………………………….…...…46 Figure 14. GALNT1 regulates EGFR O-glycosylation…………………………………47 Figure 15. Knockdown of GALNT1 enhances EGFR co-localization with EEA1 and LAMP1………………………………………………………………………48 Figure 16. Functional maps of differentially expressed genes upon GALNT1 knockdown in HCC cells……………………………………………………50 Figure 17. Quantitative RT-PCR validation of microarray results……………………...52 Table of Tables Table 1. HCC patient information…………………………………………………….53 Table 2. Differential expression of selected genes in GALNT1 knockdown HA22T and PLC5 cells………………………………………………………………54 Table 3. Real-time RT-PCR primer sequences………………………………………..55 | |
dc.language.iso | en | |
dc.title | 黏液蛋白醣化酶 N-acetylegalactosaminyltransferase 1 (GALNT1) 在肝細胞癌扮演之角色 | zh_TW |
dc.title | The role of N-acetylegalactosaminyltransferease 1 (GALNT1) in hepatocellular carcinoma | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-1 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 吳耀銘 | |
dc.contributor.oralexamcommittee | 賴逸儒,劉雅雯,李明學 | |
dc.subject.keyword | 肝細胞癌,黏液蛋白醣化?, | zh_TW |
dc.subject.keyword | hepatocellular carcinoma,GALNT1,EGFR,O-glycosylation, | en |
dc.relation.page | 55 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-01-28 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 解剖學暨細胞生物學研究所 | zh_TW |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
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