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系統識別號 U0026-2208201612492500
論文名稱(中文) 研究口腔癌細胞和免疫細胞之間的互動關係
論文名稱(英文) Study the dialogue between oral cancer cells and immune cells
校院名稱 成功大學
系所名稱(中) 分子醫學研究所
系所名稱(英) Institute of Molecular Medicine
學年度 104
學期 2
出版年 105
研究生(中文) 黃俞瑄
研究生(英文) Yu-Hsuan Huang
學號 T16034022
學位類別 碩士
語文別 英文
論文頁數 47頁
口試委員 指導教授-吳梨華
口試委員-廖楓
口試委員-趙麗洋
口試委員-凌斌
中文關鍵字 口腔癌  調節性T細胞  FOXP3  CCL22 
英文關鍵字 Oral cancer  Regulatory T cells  FOXP3  CCL22 
學科別分類
中文摘要 在台灣口腔癌已成為第五大致死癌症。大部分的口腔癌患者有伴隨著免疫失調的現象。有研究顯示誘導調節性T細胞和骨髓衍生抑制細胞的浸潤對於腫瘤逃離宿主的免疫監督為重要的調控機制。因此為了深入探討在口腔癌癌過程中,對於骨髓衍生抑制細胞或是調節性T細胞的召集作用,使用動物實驗餵食老鼠4-NQO和檳榔鹼這兩種藥物來誘發口腔癌的發生,並且利用流式細胞儀分析口腔癌過程中每個月時間點老鼠血液中這兩種免疫細胞的含量。結果發現在餵食藥物組別中,只有調節性T細胞在每個時間點的含量皆高於對照組。除了在餵食藥物組的舌頭組織,鄰近頸部的淋巴結也有顯著地增加調節性T細胞的累積。已知有許多癌症腫瘤細胞分泌的CCL22會影響調節性T細胞的累積。我們發現在餵食藥物組中CCL22的高度表現並且在68位口腔癌患者的組織中發現調節性T細胞的指標FOXP3與CCL22之間表現一致增加。Kaplan-Meier存活分析顯示相較於CCL22表現量低的口腔癌患者,高度表現CCL22的患者可能會伴隨較差的預後。腫瘤微環境中促進發炎的細胞激素TNF-或IL-可以促進腫瘤細胞CCL22產生的一種機制分別在口腔癌老鼠實驗和細胞實驗中看到。腫瘤細胞釋出的CCL22不僅會透過旁泌作用來調節有表現CCL22受體CCR4的T細胞移動,也會透過自泌作用調控口腔癌細胞的增生和移動能力。儘管先前研究已知CCL22會召集調節性T細胞,然而仍需探討在口腔癌發展過程中,高度表現的CCL22參與在口腔癌細胞和調節性T細胞之間的作用機制。
英文摘要 Oral cancer is the 5th leading cause of cancer death in Taiwan. Most of these patients demonstrate compromised immunity. The induction of regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) infiltration are important mechanisms for the tumor escape from immunological surveillance. To further investigate the recruitment of MDSCs and/or Tregs during oral cancer progression in vivo, mice were co-treated with 4-NQO and arecoline for oral cancer induction and their blood was monthly collected for flow cytometry. The percentage of circulating Tregs but not MDSCs was significantly increased at all the time points during drug-induced oral cancer progression. In addition to the increased FOXP3 expression in affected tongue tissues, we also detected the increased infiltration of Tregs in cervical lymph nodes relative to that in control animals. Since tumor-derived CCL22 is responsible for the accumulation of Tregs in certain cancer types, we also detected a significant increase of CCL22 in drug-treated tongue tissues and a concordant increase of both CCL22 and FOXP3 expression in 68 oral cancer patients. Kaplan-Meier survival curve analysis showed that high CCL22 patients tended to have poor clinical outcome related to those in low groups. The expression of inflammatory cytokines including IL-1 and TNF- in tumor microenvironment was one mechanism responsible for the increased CCL22 expression in oral cancer mouse model and oral cancer cells. Intratumor expression of CCL22 was not only functional in mediating CCR4-expressing T cell migration through paracrine release but also regulating oral cancer proliferation and migration in an autocrine manner. Although CCL22 was previously shown to recruit Tregs, more studies are needed to address the mechanisms responsible for the increased CCL22 in oral cancer and its function involved in the crosstalk of oral cancer cells and Tregs during oral carcinogenesis.
論文目次 CHINESE ABSTRACT I
ABSTRACT II
ACKNOWLEDGEMENTS III
CONTENT IV
TABLES AND FIGURES VII
ABBREVIATIONS VIII

I. INTRODUCTION 1
1-1 oral cancer 1
1-2 Tumor microenvironment 1
1-3 The role of MDSCs in cancer progression 2
1-4 The role of Tregs in cancer progression 3
1-5 CCL22, a chemotactic cytokine for Tregs 3
II. HYPOTHESIS 5
III. SPECIFIC AIMS 6
IV. MATERIALS AND METHODS 7
4-1 Cell lines 7
4-2 Oral cancer patient samples 7
4-3 Animals 7
4-4 Drug-induced oral cancer mouse model 8
4-5 Tissue collection 8
4-6 Isolation of peripheral blood mononuclear cells, spleenocytes and cervical lymph node cells 9
4-7 Flow cytometry 9
4-8 Hematoxylin and eosin staining 10
4-9 Quantitative reverse transcription and polymerase chain reaction 10
4-10 Conditioned medium collection 10
4-11 Doubling time 10
4-12 Wound-healing assay 11
4-13 Enzyme-linked immunosorbent assay for CCL22 11
4-14 Chemotaxis assay 11
4-15 Statistical analysis 12
V. RESULTS 13
5-1 The induction of oral lesions, reminiscent of SCC, by 4-NQO and arecoline 13
5-2 MDSCs and Tregs were differentially recruited during drug-induced oral cancer progression and Tregs might play a role in oral cancer progression 13
5-3 Tregs-associated genes were significantly induced during oral cancer induction and CCL22 expression was positively correlated with FOXP3 expression 14
5-4 The increase of CCL22 expression was associated with poor prognosis in oral cancer patients 15
5-5 Inflammatory cytokines induced CCL22 production during oral cancer progression in vivo and in vitro 16
5-6 Recombinant CCL22 or CCL22-bearing CM induced Jurkat T cells migration 16
5-7 Intratumoral CCL22 depletion reduced oral cancer cells proliferation and migration 17
5-8 Increased release of CCL22 in oral cancer patient sera 17
VI. DISCUSSION 18
VII. REFERENCES 22
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