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研究生:呂雅苓
研究生(外文):Ya-Ling Lu
論文名稱:茄紅素可促進PPARγ-LXRα-ABCA1之途徑而抑制男性荷爾蒙非依賴型前列腺癌細胞之生長
論文名稱(外文):Lycopene-induced growth inhibition of androgen-independent human prostate cancer cells in vitro via PPARγ-LXRα-ABCA1 pathway
指導教授:胡淼琳胡淼琳引用關係
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品暨應用生物科技學系
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:112
中文關鍵詞:茄紅素PPARLXRABCA1前列腺癌細胞膽固醇運輸
外文關鍵詞:LycopenePPARLXRABCA1prostate cancercholesterol efflux
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類胡蘿蔔素(Carotenoids),包含茄紅素(lycopene)與β-胡蘿蔔素(β-carotene),在生理上具有許多重要功能,包括:光保護作用(photoprotection)、抗氧化作用、免疫調節作用(immunomodulatory)以及抗癌活性(anti-cancer)。許多研究指出攝取富含茄紅素的食物可以降低多種癌症的發生率,如前列腺癌、肺癌等。然而茄紅素在抑制前列腺癌生長的機制目前尚未明確。因此本研究目標即在探討茄紅素在抑制前列腺癌生長中所扮演的調控角色與機制為何。本研究提出一假說,亦即:茄紅素可促進peroxisome proliferator-activated receptor gamma (PPARγ)和liver X receptor alpha (LXRα)之途徑,繼而增加下游基因ATP-binding cassette transporter A1(ABCA1)的表現,而提高細胞內膽固醇之輸出,進而抑制男性荷爾蒙非依賴型前列腺癌細胞的生長。

本研究以茄紅素不同濃度(2.5、5、10、20 μM)分別處理男性荷爾蒙非依賴型前列腺癌細胞(DU145、PC-3)。結果顯示,茄紅素會造成細胞內的膽固醇量減少,而medium中的游離膽固醇量增加,且PPARγ、LXRα和ABCA1的表現量隨著茄紅素的濃度增加(2.5 μM至10 μM)而增加,呈濃度效應。以10 μM茄紅素與DU145細胞培養12、24及48小時,隨著時間增加(12小時到24小時),膽固醇運輸量增加;同時PPARγ、LXRα和ABCA1的表現量也跟著上升。以前列腺癌細胞分別處理PPARγ(GW9662)、LXRα(GGPP)專一性的拮抗劑時,會顯著阻礙茄紅素誘導的膽固醇運輸,也降低了ABCA1的蛋白質和mRNA表現量。

總合以上結果,我們證實了茄紅素可藉由PPARγ- LXRα-ABCA1路徑促進膽固醇運輸,而抑制男性荷爾蒙非依賴型前列腺癌細胞的生長。因此,本研究結果可提供茄紅素在預防和治療前列腺癌的新理論觀念。
Carotenoids including lycopene and β-carotene possess several common biological functions such as photoprotection, antioxidant effects, and immunomodulatory and anticancer activity. Studies have suggested that higher intakes of lycopene are associated with a reduced risk of several types of cancer, such as prostate cancer, hepatoma and coronary heart disease. Although lycopene has been shown to inhibit proliferation, its mechanism of action is poorly understood. Here, we used androgen-independent human prostate cancer cells to test whether lycopene can inhibit the growth of prostate cancer via up-regulation of the expression of liver X receptor-target gene. The antiproliferative activity of lycopene may be due to LXR signaling.
Androgen-independent human prostate cancer cells (DU145、PC-3) were incubated with different concentration of lycopene (2.5, 5, 10, 20μM), and the free cholesterol in medium increased but total intracellular cholesterol decreased. At the same time, the expression of PPARγ, LXRα, ABCA1 was enhanced in a dose-dependent manner from 2.5μM to 10μM. The treatment of prostate cancer cells with 10μM lycopene for 12, 24 and 48 h promoted cellular cholesterol efflux in a time-dependent manner from 12 h to 24 h, meanwhile expression of PPARγ, LXRα, ABCA1 was also raised respectively. Addition of different specific antagonists of PPARγ(GW9662),LXRα (GGPP) to prostate cancer cells significantly suppressed lycopene-induced cholesterol efflux. In addition treatment with specific inhibitors of PPARγ orLXRα decreased lycopene-induced ABCA1 mRNA and protein expression.
The present results indicate that lycopene promotes cholesterol efflux from Androgen-independent human prostate cancer cells, and the underlying mechanism of this action is PPARγ-LXRα-ABCA1 dependent pathway. These results suggest that the effect of lycopene on proliferation may be mediated through the liver X recetor signaling pathway.
表次………………………………………………………………………Ⅴ
圖次………………………………………………………………………Ⅵ
縮寫表……………………………………………………………………Ⅸ
文獻整理……………………………………………………………… 1
一、前列腺癌………………………………………………………… 1
(一) 前列腺癌的流行病學………………………………………… 1
(二) 前列腺癌的病源學…………………………………………… 1
(三) 前列腺癌的治療方法………………………………………… 3
二、茄紅素 (lycopene)……………………………………………… 5
(一) 茄紅素簡介…………………………………………………… 5
(二) 茄紅素結構…………………………………………………… 5
(三) 茄紅素功用…………………………………………………… 6
(四) 茄紅素與攝護腺癌…………………………………………… 7
(五) 茄紅素的抗癌機制…………………………………………… 8
三、核受體與前列腺癌……………………………………………… 9
(一) 核受體………………………………………………………… 9
(二) Peroxisome proliferator-activated receptor (PPAR) 11
(三) PPARγ與前列腺癌………………………………………… 12
(四) Liver X receptor (LXR ) ……………………………… 12
(五) LXR致效劑(agonist)與前列腺癌………………………… 13
(六) PPARγ與LXRα的表現……………………………………… 15
(七) 類胡蘿蔔素、核受體、前列腺癌……………………………15
(八) 膽固醇與前列腺癌………………………………………… 17
研究動機……………………………………………………………… 20
研究目的與假說……………………………………………………… 20
實驗架構……………………………………………………………… 21
材料與方法…………………………………………………………… 22
一、實驗材料………………………………………………………… 22
二、實驗方法………………………………………………………… 25
(一) 茄紅素製備………………………………………………… 25
(二) 細胞培養…………………………………………………… 25
(三) MTT assay…………………………………………………… 27
(四) Western blotting………………………………………… 28
(五) 總RNA之萃取……………………………………………… 31
(六) 反轉錄作用………………………………………………… 32
(七) 聚合酶連鎖反應(PCR) …………………………………… 32
(八) DNA電泳……………………………………………………… 33
(九) 總膽固醇之測定…………………………………………… 33
(十) 統計分析…………………………………………………… 34
(十一) 相乘效應之計算…………………………………… …… 35
結果…………………………………………………………………… 36
一、茄紅素對DU145及PC-3前列腺癌細胞生長之抑制效力……… 36
二、茄紅素對DU145及PC-3前列腺癌細胞中PPARγ、LXRα、
ABCA1及SREBP1蛋白質表現影響…………………………………… 36
三、茄紅素對DU145前列腺癌細胞中PPARγ、LXRα、ABCA1及
SREBP1 mRNA表現之影響…………………………………………… 39
四、茄紅素對DU145前列腺癌細胞內及medium中膽固醇含量之影響41
五、PPARγ、LXRα拮抗劑對茄紅素作用之DU145前列腺癌細胞
生長抑制效力之影響……….……………………………………… 42
六、PPARγ、LXRα拮抗劑對茄紅素作用之DU145前列腺癌細胞中
PPARγ、LXRα、ABCA1、SREBP1蛋白質表現之影響……………… 42
七、PPARγ、LXRα拮抗劑對茄紅素作用之DU145前列腺癌細胞中
PPARγ、LXRα、ABCA1、SREBP1 mRNA表現之影響…………………43
八、PPARγ、LXRα拮抗劑對茄紅素作用之DU145前列腺癌細胞內及
medium中膽固醇含量之影響………………………………………… 44
九、茄紅素本身以及合併TO901317對DU145前列腺癌細胞中
PPARγ、LXRα、ABCA1、SREBP1蛋白質表現之影響……………… 44
討論…………………………………………………………………… 46
結論…………………………………………………………………… 51
圖表…………………………………………………………………… 52
參考文獻……………………………………………………………… 99
附錄…………………………………………………………………… 113
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