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系統識別號 U0026-0709201013591300
論文名稱(中文) 探討Oct4在肺癌中調控VCC-1的角色與機制
論文名稱(英文) Roles of Oct4 in regulation of VCC-1 in lung cancer
校院名稱 成功大學
系所名稱(中) 微生物及免疫學研究所
系所名稱(英) Department of Microbiology & Immunology
學年度 98
學期 2
出版年 99
研究生(中文) 陳奐勻
研究生(英文) Huan-Yun Chen
學號 S4697101
學位類別 碩士
語文別 中文
論文頁數 45頁
口試委員 口試委員-吳昭良
口試委員-李哲欣
口試委員-張芝瑞
口試委員-張孟雅
指導教授-蕭璦莉
中文關鍵字 肺癌 
英文關鍵字 VCC-1  Oct4  CXCL17 
學科別分類
中文摘要 胚胎發育 (embryogenesis) 與腫瘤形成 (oncogenesis)有許多共同的特性,Oct4為胚胎發育過程中重要的轉錄因子,調控胚胎早期發育的基因表現及影響胚胎細胞分化。在先前我們的研究發現在Oct4 (Octamer 4)表現量與腫瘤轉移及惡化程度成正比,且透過DNA微陣列分析發現當Oct4表現提高,VEGF-correlated chemokine-1 (VCC-1)表現亦升高七倍。在本研究中首先於臨床肺癌檢體的組織染色分析結果證實Oct4表現與VCC-1表現呈正相關;在293T 人類胚胎細以及H1299和A549肺癌細胞大量表現Oct4或抑制Oct4表現後,以反轉錄聚合酶連鎖反應與免疫墨點分析證明VCC-1表現隨Oct4表現提高或下降。進一步構築VCC-1 啟動子並進行報導基因表現分析,發現Oct4可藉由與VCC-1啟動子結合,進而調控VCC-1表現。由細胞移動分析試驗結果發現, H1299肺癌細胞分泌的VCC-1,可專一性吸引經 phorbol 12-myristate 13-acetate (PMA)活化的THP-1類巨噬細胞。當H1299大量表現VCC-1同時,可促進肺癌細胞大量分泌乙型轉型生長因子(transforming growth factor-β, TGF-β)。此外,將抑制VCC-1表現的H1299肺癌細胞,以背部皮下注射至小鼠,觀察腫瘤生長情形,發現VCC-1表現量低的處理組,腫瘤大小亦相對較小。綜合以上結果推論VCC-1可促進腫瘤生長及惡化,然而目前對於VCC-1在腫瘤中所扮演的角色仍待進一步釐清。此外VCC-1或許可作為肺癌初期的腫瘤標記及治療肺癌的標的基因。
英文摘要 Oct4 (Octamer 4) is a member of the POU family of transcription factors, which is expressed in pluripotent human embryonic stem (ES) cells. Oct4 is required to sustain self-renewal ability of ES cells. Recent studies have shown expression of Oct4 in some tumor tissues and cell lines, supporting the stem cell theory of carcinogenesis. We previously reported that overexpression of Oct4 enhances, whereas knockdown of Oct4 expression reduces, migration and invasion of bladder cancer cells. VEGF-correlated chemokine 1 (VCC-1), which is also known as dendritic cell (DC)- and monocyte-attracting chemokine-like protein (DMC), is a novel cytokine-like protein which attracts DCs and monocytes. Our microarray data revealed that expression of VCC-1 was increased by 7-fold in human bladder cancer cells overexpressing Oct4 compared with their control counterparts. However, the functions of VCC-1 in tumorigenesis remain poorly understood. In this study, I investigated the role and significance of the relationship between Oct4 and VCC-1 in lung cancer. My results showed that overexpression of Oct4 increased VCC-1 expression in H1299 lung cancer cells, whereas knockdown of Oct4 reduced its expression in A549 lung cancer cells. Reporter assays also revealed that Oct4 enhanced the promoter activity of VCC-1. These results suggest that VCC-1 is one of the downstream molecules of Oct4. Furthermore, overexpression of Oct4 and VCC-1 enhanced the production of transforming growth factor-β (TGF-β) by lung cancer cells, suggesting that TGF-β may be regulated by Oct4 and VCC-1. In addition, knockdown of VCC-1 in lung cancer cells reduced tumor growth in NOD/SCID mice bearing subcutaneous H1299 tumors. Collectively, this study may not only elucidate the functions of Oct4 and VCC-1 and their correlation in lung cancer, but also implicate their usefulness as tumor prognostic markers and therapeutic targets.
論文目次 中文摘要...................................... I
Abstract III
誌謝………………………………………………………………………V
目錄 ………………………………………………………………… .VI
圖目錄..................................... IX
縮寫與符號.................................... X
緒論 1
1. 肺癌 (lung cancer)................ 1
2. Oct4 (Octamer-4)................. 2
2.1. Oct4與癌症的關聯性..................... 2
2.2. Oct4 背景......................... 3
2.3. Oct4調控的下游基因 ........................3
3. VCC-1 (VEGF co-regulation chemokine-1) 4
3.1 . VCC-1背景.......................... 4
3.2 VCC-1與癌症的關聯性...................... 5
研究目的與試驗設計................................ 6
材料與方法................................... 7
A. 材料............................... 7
A.1.質體(Plasmids)........................... 7
A.2.細胞株................................... 8
A.3.核酸片段............................... 9
A.4.實驗動物............................... 10
A.5.臨床檢體搜集............................. 10
B.試驗方法.................................. 10
B.1.基因之蛋白質表現量分析.................... 10
B.3.基因轉染試驗 (Gene transfection)............. 12
B.5.於肺癌細胞中分析VCC-1 啟動子活性 (Analysis of VCC-1 promoter activities in lung cancer cell lines)......... 15
B.6.細胞移動分析 (migration assay)............ 16
B.7.酵素聯結免疫吸附分析 ( Enzyme-linked immunosorbent assay, ELISA)..................................... 17
B.8.動物試驗 (Animal studies).............. 18
B.9.統計分析 (Statistical analysis)......... 19
結果 …………………………………………………………………..20
1. VCC-1 與 Oct4 的表現量隨肺癌惡化程度提高而增加 20
2. 在肺癌細胞中 Oct4 表現與 VCC-1 呈正相關...... 21
3. Oct4經由調控VCC-1轉錄作用進而影響VCC-1表現... 22
4. 肺癌細胞可藉由分泌VCC-1進而吸引macrophage-like THP-1細胞 23
5. 肺癌細胞分泌VCC-1可促進TGF-β表現量增加....... 24
6. 肺癌細胞大量表現VCC-1不影響VEGF分泌.... 24
討論 …………………………………………………………………..26
參考文獻.................................. 31

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