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系統識別號 U0026-0709201014145900
論文名稱(中文) 長鏈脂肪酸延長酶 6 對肝癌細胞的影響
論文名稱(英文) Effects of ELOVL6 on Liver Cancer
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) of Biochemistry and Molecular Biology
學年度 98
學期 2
出版年 99
研究生(中文) 黃耀生
研究生(英文) Yaw-Sheng Huang
學號 S1697104
學位類別 碩士
語文別 中文
論文頁數 45頁
口試委員 指導教授-吳昭良
口試委員-賴明德
口試委員-蕭璦莉
口試委員-張芝瑞
口試委員-張孟雅
中文關鍵字 長鏈脂肪酸延長酶 6  肝癌 
英文關鍵字 ELOVL6  HCC 
學科別分類
中文摘要 肝癌為遍布世界各地的疾病,特別在亞洲地區,肝癌更屬於好發性且預後情況較差的癌症。目前已知代謝異常是肝癌形成的危險因子之一。長鏈脂肪酸延長酶 6 (elongation of long chain fatty acids family member 6, ELOVL6) 為一種脂肪酸延長酶,專一作用在延長 12、14 及 16 碳的飽和脂肪酸與單元不飽和脂肪酸。因此,ELOVL6 在脂質合成活躍的器官或組織,例如肝臟和脂肪中有較高的表現量。有研究指出,參與脂質合成的酵素在肝癌中較為活化,並且 ELOVL6 的表現也高於正常組織。在本研究中,主要探討 ELOVL6 對於肝癌細胞生長的影響。我們發現利用腺病毒攜帶核醣核酸干擾因子抑制 ELOVL6 的表現,可抑制小鼠肝癌細胞的增生,並且這種抑制機制主要為使細胞週期停滯在 G1 期。當 ELOVL6 的表現量降低後,發現細胞內的脂質代謝作用,包含脂質的堆積與氧化現象有增加的情形,此脂質代謝的改變可能由細胞內 16 碳脂肪酸的比例增加造成。由小鼠皮下肝腫瘤模式中,我們發現抑制 ELOVL6 的表現可以抑制腫瘤的生長。綜合以上結果,我們在細胞與動物實驗中抑制 ELOVL6 的表現,皆可抑制肝癌細胞的生長。因此,利用腺病毒攜帶 ELOVL6 的核醣核酸干擾因子在肝癌的治療上,可作為一個具有潛力的方法。
英文摘要 Hepatocellular carcinoma (HCC) is one of the most prevalent cancers worldwide, particularly in Asia. The prognosis of HCC is poor. Metabolic dysregulation is one of the risk factors for the development of HCC. Elongation of long chain fatty acids family member 6 (ELOVL6), also known as LCE and FACE, is a long chain fatty acid elongase that specifically catalyzes the elongation of saturated and monounsaturated fatty acids with 12, 14, and 16 carbons. ELOVL6 is highly expressed in the liver and adipose tissue where lipogenesis is active. Previous studies have shown that lipogenic enzymes, including ELOVL6, were activated in HCC. In this study, we aimed to investigate the effects of ELOVL6 on the growth of HCC. We found that knockdown of ELOVL6 expression in ML-1 murine hepatoma cells induced morphological changes and growth inhibition due to cell cycle arrest at the G1 phase. In addition, knockdown of ELOVL6 in ML-1 cells increased lipid accumulation and β-oxidation, suggesting that downregulation of ELOVL6 results in cell cycle arrest through the alteration of lipid metabolism. Furthermore, tumor growth was suppressed by adenovirus-mediated ELOVL6 gene silencing through RNA interference in a subcutaneous ML-1 tumor model. Altogether, our in vitro and in vivo results suggest that adenovirus-mediated ELOVL6 gene silencing may have therapeutic potential for the treatment of liver cancer.
論文目次 中文摘要 II
英文摘要 III
誌謝 IV
目錄 V
圖目錄 VIII
縮寫 IX
緒論 1
一、肝癌 1
二、脂質生合成 3
三、長鏈脂肪酸延長酶 6 4
研究動機 7
材料與方法 8
一、材料 8
1、細胞株 8
2、引子 8
3、腺病毒 9
4、脂肪酸 9
5、實驗動物 9
二、方法 9
1、細胞培養 9
2、腺病毒生產 10
3、腺病毒效價偵測 11
4、西方墨點法 11
5、反轉錄酶聚合連鎖反應 13
6、即時半定量反轉錄酶聚合連鎖反應 14
7、WST-1及WST-8分析 14
8、TUNEL分析 15
9、Monodansylcadaverine 染色 15
10、Propidium iodide 染色 16
11、油紅染色 16
12、BODIPY stain 16
13、動物實驗 17
14、統計分析 18
結果 19
一、ELOVL6在肝癌中有過量表現的情形 19
二、Ad.shELOVL6 可有效降低 ELOVL6 的表現 19
三、降低 ELOVL6 的表現會使肝癌細胞的生長受到抑制 19
四、降低 ELOVL6 的表現會使肝癌細胞的生長週期停滯在 G1 期 20
五、16 碳脂肪酸 palmitate 和 palmitoleate 相較於 18 碳脂肪酸 stearate 和 oleate 對肝癌細胞生長的影響較大 21
六、降低 ELOVL6 的表現會使肝癌細胞的脂質代謝產生改變 23
七、在小鼠皮下腫瘤模式中,降低腫瘤 ELOVL6 的表現可抑制腫瘤生長 23
討論 25
參考文獻 29
圖表 33
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