雄性素依賴型和非依賴型前列腺癌細胞生長和SOCS基因的調節

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吳健嘉(Chien-chia Wu) 查詢紙本館藏

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生命科學系

論文名稱

雄性素依賴型和非依賴型前列腺癌細胞生長和SOCS基因的調節
(Regulation of growth and suppressor of cytokine signaling gene in the androgen-dependent and androgen-independent prostate cancer cells)

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摘要(中)

本論文要驗證的假說有胰島素阻抗素和瘦素會差異性調節雄性素依賴型LNCaP-104S和雄性素非依賴型LNCaP-104R1、LNCaP-104R2和PC-3前列腺癌細胞的生長，另一個假說是第一型內皮素會刺激人類前列腺癌細胞SOCS基因表現。結果顯示處理48小時，胰島素阻抗素會刺激LNCaP-104R1、LNCaP-104R2和PC-3而不是LNCaP-104S前列腺癌細胞的增生 (如同增加細胞數目所指示)。瘦素則會刺激LNCaP-104S、LNCaP-104R1和PC-3前列腺癌細胞的增生。這些結果支持我們第一個假說。有趣的是，胰島素阻抗素和瘦素下游SOCS基因表現量在LNCaP-FGC和PC-3細胞有所不同。即使SOCS-1與SOCS-3在LNCaP-FGC細胞是不存在的，所有SOCS家族成員，例如SOCS-1，-2，-3，-4，-5，-6，-7和CIS-1，存在於PC-3細胞和U-937淋巴癌細胞。SOCS-3也是不存在於LNCaP-104S、LNCaP-104R1和LNCaP-104R2細胞。藉由PC-3細胞，我們發現第一型內皮素會促進SOCS-1，-3和-5 mRNA表現量隨著時間和劑量的不同而有所差異並且對其它SOCS家族成員的mRNA量則是沒有作用。除此之外，單獨處理綠茶唲茶素會促進PC-3細胞SOCS-1，-3與-5 mRNA量並且不改變其它SOCS家族成員的mRNA表現。在第一型內皮素存在的情況下，有趣的是發現EGCG會抑制第一型內皮素誘導PC-3細胞SOCS-1，-3與-5 mRNA量。這些研究成果認為EGCG和第一型內皮素會選擇性影響特定SOCS家族成員的種類。就如同部分SOCSs已被證實會調控前列腺癌細胞的生長，這些研究成果也許能解釋脂肪激素和第一型內皮素誘導前列腺癌細胞生長，以及EGCG抑制前列腺癌細胞生長的機制。

摘要(英)

The present thesis was to test the hypothesis that resistin and leptin differentially regulated the growth between androgen-dependent LNCaP-104S and androgen-independent LNCaP-104R1、LNCaP-104R2 and PC-3 prostate cancer cells, and the hypothesis that endothelin (ET)-1 stimulated the gene expression of suppressors of cytokine signaling (SOCSs) in human prostate cancer cells. At treating 48 hours, resistin generally induced the proliferation (as indicated by an increased number of cells) of LNCaP-104R1、LNCaP-104R2 and PC-3 rather than LNCaP-104S prostate cancer cells. Leptin induced the proliferation of LNCaP-104S、LNCaP-104R1 and PC-3 prostate cancer cells. These data support our first hypothesis. Interestingly, the expression level of the resistin and leptin downstream SOCS genes were different between LNCaP-FGC and PC-3 cells. Although SOCS-1 and SOCS-3 were absent in the LNCaP-FGC cell, all of SOCS family members, such as SOCS-1, -2, -3, -4, -5, -6, and -7 and CIS-1, present in PC-3 cell and U-937 lymphoma cell. The SOCS-3 was also absent in LNCaP-104S、LNCaP-104R1 and LNCaP-104R2 cells. Using PC-3 cell, we found that ET-1 stimulated increases in levels of SOCS-1, -3, and -5 mRNAs in time and dose-dependent manners and had no effects on mRNA levels of other SOCS family members. In addition, treatment of PC-3 cell with green tea epigallocatechin-3-gallate (EGCG) alone induced increases in levels of SOCS-1, -3 and -5 mRNAs and unaltered mRNA expression of other SOCS family members. In the prescence of ET-1, EGCG was interestingly found to inhibit the ET-1 stimulated increases of SOCS-1, -3 and -5 mRNA levels in PC-3 cell. These data suggest that EGCG and ET-1 selectively affects particular types of SOCS family members. As some of SOCSs have been reported to regulate growth of prostate cancer cells, results of this study may help explain the mechanism in the adipokine and ET-1 stimulation of prostate cancer cell growth, as well as the mechanisms of which EGCG suppresses growth of prostate cancer cells.

關鍵字(中)

★ 胰島素阻抗素
★ 瘦素
★ 第一型內皮素
★ 前列腺癌細胞

關鍵字(英)

論文目次

中文摘要 …………………………………………………………………………Ⅴ
英文摘要 …………………………………………………………………………Ⅵ
誌謝 ………………………………………………………………………………Ⅷ
目錄 ………………………………………………………………………………Ⅸ
表目錄 ……………………………………………………………………………XI
圖目錄 ……………………………………………………………………………XII
縮寫與全名對照 ………………………………………………………………XIII
第一章、緒論 ………………………………………………………………………1
1. 前列腺癌 (Prostate cancer)………………………………………………1
2. 胰島素阻抗素 (Resistin)…………………………………………………2
3. 瘦素 (Leptin)………………………………………………………………3
4. 第一型內皮素 (Endothelin-1)……………………………………………3
5. SOCS蛋白質 (Suppressor of cytokine signaling)……………………4
6. 研究動機與目的 ……………………………………………………………5
第二章、實驗材料與方法…………………………………………………………7
1. 材料 …………………………………………………………………………7
2. 細胞培養 ……………………………………………………………………7
3. BrdU滲入法 …………………………………………………………………9
4. RNA萃取 (RNA extraction) ………………………………………………9
5. 反轉錄反應 (Reverse Transcription) …………………………………10
6. 聚合酶鏈反應 (Polymerase Chain Reaction，PCR) ……………………11
7. PCR產物分析 ………………………………………………………………12
8. Real-time PCR (Q-PCR) ……………………………………………………12
9. 統計分析………………………………………………………………………13
第三章、結果………………………………………………………………………14
1. 胰島素阻抗素影響不同型人類前列腺癌細胞株的生長……………………14
2. 瘦素影響不同型人類前列腺癌細胞株的生長………………………………15
3. 以BrdU assay分析胰島素阻抗素對不同型人類前列腺癌細胞株增生的影響………………………………………………………………………………16
4. 以BrdU assay分析瘦素對不同型人類前列腺癌細胞株增生的影響………17
5. SOCS家族基因存在於人類前列腺癌細胞株…………………………………17
6. 第一型內皮素對PC-3前列腺癌細胞內SOCS基因之影響…………………18
7. EGCG會抑制第一型內皮素對PC-3前列腺癌細胞內SOCS基因之表現……18
第四章、討論………………………………………………………………………20
1. 胰島素阻抗素會差異性調節不同型人類前列腺癌細胞株之生長………………………………………………………………………………20
2. 瘦素會差異性調節不同型人類前列腺癌細胞株之生長……………………21
3. PC-3雄性素非依賴型與LNCaP-FGC雄性素依賴型前列腺癌細胞，SOCS基因家族是否存在…………………………………………………………………22
4. 第一型內皮素促進PC-3前列腺癌細胞部分SOCS基因表現，具有時間和劑量效應…………………………………………………………………………22
5. EGCG會影響第一型內皮素促進PC-3前列腺癌細胞內SOCS基因表現……23
第五章、結論………………………………………………………………………26
第六章、參考文獻…………………………………………………………………27
第七章、圖表………………………………………………………………………35
第八章、附錄………………………………………………………………………53

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指導教授

高永旭(Yung-Hsi Kao)

審核日期

2014-4-17

推文