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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 | Mei-Ieng Che | en |
dc.contributor.author | 謝美英 | zh_TW |
dc.date.accessioned | 2021-06-16T06:33:44Z | - |
dc.date.available | 2014-10-09 | |
dc.date.copyright | 2014-10-09 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-08-04 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57051 | - |
dc.description.abstract | β1,4-N-acetylgalactosaminyltransferase III (B4GALNT3) 具有轉移GalNAc到所有N-glycans 和 O-glycans末端GlcNAc-β的功能,是一個合成GalNAcβ1-4GlcNAc (LacdiNAc) 醣結構的酵素。在我們過去的研究發現,過度表現B4GALNT3能增加大腸直腸癌細胞的惡性程度,但其作用機轉並不明確。我們發現與正常的大腸比較,B4GALNT3在大腸直腸癌的表現量是增加,而且與病人較差的存活率有關。在大腸直腸癌細胞株HCT116和SW480中過度表現B4GALNT3後,會增加EGF所誘導的轉移與入侵能力。抑制B4GALNT3在大腸直腸癌細胞株HCT15和HT29的表現後,會減弱EGF所誘導的轉移與入侵能力,B4GALNT3所引起的細胞爬行能力會被EGFR抑制劑erlotinib減弱。而且過度表現B4GALNT3後也會增加EGF所誘導細胞球體形成能力。反之,抑制B4GALNT3表現後,這些惡性的細胞表現型也會減弱。Wisteria floribunda agglutinin (WFA) 沉澱試驗發現,B4GALNT3的表現可以改變EGFR上N-glycans的LacdiNAc表現,增加B4GALNT3在細胞中的表現會增強活化的EGFR表現及其下游的訊息傳遞。然而,增加B4GALNT3表現後,不會對HGF所引導的遷移能力產生影響,也不影響MET的醣類結構,而且B4GALNT3的表現量對bFGF所引導的細胞球體形成能力也不會產生影響。我們的結果說明B4GALNT3能透過修飾EGFR的醣化及控制其訊息傳遞來調控癌細胞的幹性及入侵能力,我們認為抑制B4GALNT3的表現具有治療大腸直腸癌的潛力。 | zh_TW |
dc.description.abstract | β1,4-N-acetylgalactosaminyltransferase III (B4GALNT3) catalyzes the transfer of GalNAc to any nonreducing terminal GlcNAc-β on N- and O-glycans, resulting in the synthesis of GalNAcβ1-4GlcNAc (LacdiNAc) structure. Although we previously showed that B4GALNT3 overexpression increased colon cancer cell malignant phenotypes in vitro and tumorigenesis in vivo, the underlying mechanism is poorly understood. Immunohistochemistry showed that B4GALNT3 was upregulated in colorectal tumors compared with their surrounding non-tumor colorectal tissues and was associated with poor survival of colorectal cancer patients. B4GALNT3 overexpression enhanced EGF-induced migration and invasion in HCT116 and SW480 cells. Knockdown of B4GALNT3 reduced EGF-induced migration and invasion in HCT115 and HT29 cells. The effects of B4GALNT3 on migration and invasion induced by 10% FBS were blocked with erlotinib, an EGFR inhibitor. Moreover, overexpression of B4GALNT3 increased EGF-induced sphere formation, whereas B4GALNT3 knockdown reduced cell malignant phenotypes. Wisteria floribunda agglutinin (WFA) pull down assays showed that B4GALNT3 primarily decorated the LacdiNAc structure on the N-glycans of EGFR. B4GALNT3 overexpressing cells enhanced EGF-induced phosphorylation of EGFR and its downstream signaling pathway. Overexpression of B4GALNT3 in colon cancer cells altered neither the HGF-induced motility and glycosylation of MET nor the bFGF-induced sphere forming ability. Our results suggest that B4GALNT3 regulates cancer stemness and the invasive properties of colon cancer cells through modifying EGFR glycosylation and signaling. Our findings provide a novel strategy for colorectal cancer therapy by inhibiting B4GALNT3. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T06:33:44Z (GMT). No. of bitstreams: 1 ntu-103-F96446002-1.pdf: 1912595 bytes, checksum: ce7cca72bfef17a326e8f7c20b6e4710 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | Table of contents
Chapter 1. Introduction 6 1.1. Growth factor receptors and treatment for colorectal cancer 6 1.2. Protein glycosylation 8 1.3. Glycosylation and its relationship with receptor tyrosine kinases 9 1.4. β1,4-N-acetylgalactosaminyltranferases (β4GalNAc-Ts) 10 Chapter 2. Specific aims 12 Chapter 3. Materials and methods 13 3.1. Patients 13 3.2. Immunohistochemistry 13 3.3. Cell culture 14 3.4. Stable expression of B4GALNT3 14 3.5. Knockdown of B4GALNT3 15 3.6. Real-time RT-PCR 15 3.7. Western blot analysis 17 3.8. Lectin pull down 17 3.9. Sphere formation assay 18 3.10. Transwell migration assay 18 3.11. Matrigel invasion assay 19 3.12. Statistical analyses 19 Chapter 4. Results 20 4.1. B4GALNT3 expression in primary colorectal tumors 20 4.2. B4GALNT3 regulates LacdiNAc expression in colon cancer cell lines 21 4.3. B4GALNT3 does not regulate HGF-induced invasive phenotypes and glycosylation of MET 22 4.4. B4GALNT3 mediates EGF-induced migration and invasion 22 4.5. B4GALNT3 modulates glycosylation of EGFR 23 4.6. B4GALNT3 regulates stem-like potential in colon cancer cells 24 4.7. B4GALNT3 modulates EGF-induced but not bFGF-induced sphere formation 25 4.8. B4GALNT3 modifies activity of EGFR 26 Chapter 5. Discussion 27 5.1. Glycosylation in cancer stem cells 27 5.2. The effect of B4GALNT3 in tumors 27 5.3. B4GALNT3 regulates the glycosylation and activity of EGFR in colon cancer cells 28 5.4. Other receptors may express the LacdiNAc structure 29 5.5 Targeting B4GALNT3 may be a novel strategy in colorectal cancer treatment 30 Chapter 6. Conclusion 31 References 32 Table of figures Figure 1. Immunohistochemistry of B4GALNT3 in human colorectal cancer. 43 Figure 2. The correlation between B4GALNT3 expression and clinicopathologic factors. 44 Figure 3. B4GALNT3 overexpression correlates with metastasis free survival. 46 Figure 4. The expression of B4GALNT3 in colon cancer cell lines. 47 Figure 5. The expression of B4GALNT3 regulates LacdiNAc expression in colon cancer cells. 48 Figure 6. Overexpression of B4GALNT3 does not alter HGF-induced migration and invasion in colon cancer cells. 49 Figure 7. B4GALNT3 overexpression does not change the LacdiNAc structure of MET. 50 Figure 8. Overexpression of B4GALNT3 induces EGF-induced migration and invasion in colon cancer cells. 51 Figure 9. Knockdown of B4GALNT3 suppresses EGF-induced migration and invasion in colon cancer cells. 52 Figure 10. B4GALNT3- induced migration and invasion are blocked by erlotinib. 54 Figure 11. B4GALNT3 overexpression increases the LacdiNAc structure mainly on N-glycans of EGFR. 55 Figure 12. Knockdown of B4GALNT3 decreases LacdiNAc structure on EGFRs. 57 Figure 13. B4GALNT3 regulates sphere formation in colon cancer cells. 58 Figure 14. B4GALNT3 modulates OCT4 and NANOG expression at mRNA and protein level inHCT116 cells. 59 Figure 15. EGF increases sphere formation in colon cancer cells. 60 Figure 16. bFGF induces sphere formation in colon cancer cells. 61 Figure 17. B4GALNT3 mediates EGF-induced sphere formation in colon cancer cells. 62 Figure 18. B4GALNT3 does not regulate sphere formation in serum free condition. 64 Figure 19. B4GALNT3 does not modulate bFGF-induced sphere formation in colon cancer cells. 65 Figure 20. Overexpression of B4GALNT3 enhance the activity of EGFR in HCT116 cells. 67 | |
dc.language.iso | en | |
dc.title | "β1,4-N-acetylgalactosaminyltransferase III 控制EGFR調控之大腸直腸癌細胞幹性" | zh_TW |
dc.title | β1,4-N-acetylgalactosaminyltransferase III modulates cancer stemness through EGFR signaling pathway in colon cancer cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 何宛玲,張修豪,黃約翰,陳?豪 | |
dc.subject.keyword | B4GALNT3,EGFR,大腸直腸癌,LacdiNAc,癌幹細胞, | zh_TW |
dc.subject.keyword | B4GALNT3,EGFR,colorectal cancer,LacdiNAc,cancer stem cells, | en |
dc.relation.page | 67 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-08-04 | |
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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