探討TIMP-3在EGFR抑制劑治療神經母細胞瘤中的調節機制

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陳婷軒(Ting-Xuan Chen) 查詢紙本館藏

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生命科學系

論文名稱

探討TIMP-3在EGFR抑制劑治療神經母細胞瘤中的調節機制
(Exploring the regulation mechanism of TIMP-3 in the EGFR inhibitor treatment of neuroblastoma)

相關論文

★ 探討化合物 Y 抑制神經母細胞瘤增生之效果	★ 探討化合物Y抑制神經母細胞瘤轉移之效果
★ 探討硒代胱氨酸於神經母細胞瘤之治療潛力

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至系統瀏覽論文 (2026-8-31以後開放)

摘要(中)

Neuroblastoma (NB) is the most common extracranial solid tumor in children, with high-risk NB patients facing a survival rate of less than 50%. Therefore, a novel therapeutic avenue for NB is imperative. Notably, EGFRs, ErbB1 and ErbB2, are highly expressed in NB cells, and activating these receptors significantly promotes the proliferation of NB cells. Targeting ErbB1 and ErbB2 may be a potential treatment for NB. The preliminary data showed that the ErbB1 and ErbB2 dual inhibitor, CL -387785, exerts notable inhibitory effects on NB cell proliferation, migration, and invasion. Additionally, transcriptomic analysis via qPCR array showed that TIMP3 was upregulated in CL-387785-treated cells. In this study, I would like to examine the therapeutic efficacy of CL-387785 in vivo and further explore the regulatory mechanism of TIMP3 in NB. By the mouse subcutaneous xenograft and tail-vein metastasis model, I found that CL-387785 effectively inhibits the tumor growth and the liver metastasis of NB cells. After EGF treatments, the mRNA expression of TIMP-3 was inhibited, while DNMT1 and EZH2 were induced. Conversely, treatments of the epigenetic regulation inhibitors, 5-Azacytidine (5-Aza), Tazemetostat (EPZ-6438), and Trichostatin A (TSA), upregulated the TIMP3 mRNA level. These in-vitro findings suggest that the EGFR pathway likely regulates TIMP3 both transcriptionally and epigenetically. However, further investigations are warranted to elucidate the precise mechanism through which EGFR modulates TIMP3 expression and consequently impacts NB progression.

摘要(英)

關鍵字(中)

★ 神經母細胞瘤
★ 表觀遺傳學

關鍵字(英)

★ Neuroblastoma
★ TIMP3
★ CL-387,785
★ EGFR signaling
★ Epigenetics

論文目次

中文摘要 i
Abstract ii
Acknowledgments iii
Table of Contents iv
List of Figures vi
List of Tables vii
I. Introduction 1
1-1 Neuroblastoma 1
1-1-1 Staging System 2
1-1-2 Treatment strategies 3
1-2 Epidermal growth factor receptor (EGFR) 4
1-3 Tissue inhibitors of metalloproteinases 3 (TIMP3) 5
1-4 Rationale 7
II. Materials and methods 8
2-1 Cell culture 8
2-1-1 SK-N-BE(2)-C 8
2-1-2 SH-SY5Y 8
2-1-3 HEK293T 8
2-2 Western blot 8
2-2-1 Cell harvesting 8
2-2-2 SDS Polyacrylamide-gel-electrophoresis (SDS-PAGE) 8
2-2-3 Protein transfer 9
2-2-4 Blocking and antiboby binding 9
2-2-5 Membrane striping 10
2-3 Real-time quantitative polymerase chain reaction (RT-qPCR) 10
2-3-1 RNA extraction 10
2-3-2 Reverse transcription 10
2-3-3 Real-time quantitative polymerase chain reaction 11
2-4 Vector Construction 11
2-4-1 PCR amplification of insert 11
2-4-2 Restriction Enzyme Digestion 12
III. Results 15
3-1 Metastasis models for examining the effect of CL-387785 on NB metastasis 15
3-2 The xenograft mouse model shows that CL inhibits NB cell proliferation. 16
3-3 EGFR treatment inhibit TIMP3 expression 16
3-4 TIMP3 expression is influenced by EZH2-mediated histone modification 16
3-5 5-Aza, EPZ-6438, TSA treatment induced TIMP3 expression . 17
3-6 Luciferase reporter assay vector construction 18
3-7 Luciferase reporter assay 18
IV. Discussion 20
V. References 24
VI. Tables and Figures 28

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指導教授

吳沛翊(Pei-Yi Wu)

審核日期

2024-8-22

推文