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系統識別號 U0026-1707201511104900
論文名稱(中文) 探討活化的巨噬細胞促進大腸癌細胞侵襲的角色
論文名稱(英文) The Role of Activated Macrophages in Enhancing the Invasive Properties of Colon Cancer Cells
校院名稱 成功大學
系所名稱(中) 生物化學暨分子生物學研究所
系所名稱(英) Department of Biochemistry and Molecular Biology
學年度 103
學期 2
出版年 104
研究生(中文) 張德沛
研究生(英文) Te-Pei Chang
學號 S16021086
學位類別 碩士
語文別 英文
論文頁數 68頁
口試委員 指導教授-吳華林
口試委員-施桂月
口試委員-林淑華
口試委員-江美治
中文關鍵字 腫瘤相關巨噬細胞  癌細胞爬行  血管新生 
英文關鍵字 Tumor-associated macrophages  Cancer cell migration  Angiogenesis 
學科別分類
中文摘要 腫瘤細胞能夠藉著一系列的細胞因子和趨化因子與在腫瘤微環境的細胞作用,以促進腫瘤細胞發展成更惡性的狀態。有研究指出,腫瘤相關巨噬細胞(TAMs)會誘導大腸癌細胞轉移。然而,關於腫瘤相關巨噬細胞的作用,許多相互矛盾的文獻致無法達成共識。因此本篇研究的目的是探討在大腸癌的腫瘤微環境中腫瘤相關巨噬細胞的功能。我們選用人類大腸癌細胞株 (SW480) 作為研究的體外模式細胞。本研究,所使用的巨噬細胞是從人類單核球細胞 (THP-1) 加入巴豆醇-12-十四烷酸酯-13-乙酸酯 (phorbol 12-myristate 13-acetate) 刺激下成為巨噬細胞 (M0),再藉由γ干擾素的刺激分化成M1型巨噬细胞 (M1),或藉由介白素-4的刺激分化成M2型巨噬细胞 (M2)。由即時聚合酶鏈鎖反應實驗結果顯示大腸癌細胞培養液刺激的M0巨噬細胞會表現高量的M1型巨噬细胞標記基因,代表大腸癌細胞會馴化巨噬細胞成為M1型巨噬细胞。為了探討M0、M1、M2型巨噬细胞培養液對大腸癌細胞爬行能力的影響,藉由細胞穿越實驗(transwell assay) ,我們發現M1型巨噬细胞培養液會吸引大腸癌細胞爬行地最多。為了探討先經由不同的巨噬细胞培養液刺激的大腸癌細胞是否會促進血管新生,我們發現先經由M1型巨噬细胞培養液刺激的大腸癌細胞的培養液會讓人類臍靜脈內皮細胞 (HUVECs) 形成比較多的管柱。此外我們也發現M1型巨噬细胞培養液刺激的大腸癌細胞並不藉由NF-κB的路徑而增加其侵襲和促進血管新生的能力。總括而言,本論文結果顯示人類大腸癌細胞會促使巨噬細胞極化成為M1型巨噬细胞進而增加大腸癌細胞的爬行能力、侵襲能力、和促進血管新生的能力但不藉由活化大腸癌細胞的NF-κB路徑。
英文摘要 Tumor cells are capable of communicating with the cells in the tumor microenvironment via a series of cytokines and chemokines to promote their progression to a more malignant state. In addition, there are studies showing that tumor-associated macrophages (TAMs) induce metastatic behavior of colon cancer cells. However, accumulated conflicting evidences regarding the role of TAMs have not yet reached a consensus. Hence, we aimed to study the functions of TAMs in the tumor microenvironment of colon cancers. We used cell line-SW480 (colorectal adenocarcinoma) in our in vitro model. In the present study, we generated TAMs from THP-1 monocytic cells stimulated with phorbol 12-myristate 13-acetate into macrophages (M0), and M0 were further differentiated into classically activated macrophages (M1) or alternatively activated macrophages (M2) by adding interferon- γ or interleukin-4 respectively. Additionally, we found that SW480 conditioned medium (CM) was able to educate M0 to become M1 that expressed high level of M1 marker genes. To examine the migration ability of SW480 cells under stimuli of M0, M1, and M2 CMs respectively, the transwell assay was performed. SW480 cells migrated more toward the M1 CM than all the other media. To explore the pro-angiogenic ability of SW480 cells with different stimulations of macrophage CM, CM obtained from M1 CM-stimulated SW480 could cause tube formation of human umbilical vein endothelial cells. In addition, we gathered the results from western blotting that stimulation of M1 CM in SW480 cells did not activate their NF-κB pathway. In summary, our data imply that SW480 cells polarize macrophages toward M1 to enhance their migration, invasion and pro-angiogeneic abilities but not by NF-κB pathway.
論文目次 Abstract in Chinese 1
Abstract in English 2
Acknowledgement 3
Content 4
Figure Contents 5
Table Contents 6
Abbreviation 7
Instruments 9
Reagents and Chemicals 11
Introduction 15
Specific Aim 19
Materials and Methods 20
Results 40
Conclusion 43
Discussion 44
References 47
Figures 55
Tables 66
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