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研究生:傅艷臻
研究生(外文):Fuh Yen-Jen
論文名稱:探討BTG2基因於前列腺癌調控之機制
論文名稱(外文):To understand the regulatory mechanisms of BTG2 gene in prostate cancer
指導教授:莊宏亨
指導教授(外文):Juang, Horng-Heng
學位類別:碩士
校院名稱:長庚大學
系所名稱:基礎醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:94
語文別:中文
論文頁數:91
中文關鍵詞:前列腺癌基因調控訊息傳遞
外文關鍵詞:BTG2NFκB53gene regulationprostate cancer
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Anti-Proliferative (APRO)基因家族中的成員B cell translocation gene 2 (BTG2)能被p53及DNA損害物質活化,藉由Rb dependent或Rb independent的途徑抑制細胞週期G1/S的進行。我們使用RT-PCR探討於不同前列腺細胞株中BTG2基因表達量,發現BTG2基因表達量的降低與前列腺癌細胞的形成有關,且將BTG2基因過度表達於PC-3癌細胞中可抑制細胞的生長。分析克隆出的BTG2基因5’-flanking區域 (-1至-297)之序列發現類似p53及NFκB的response element,皆可能與BTG2基因的調控有關。暫時性基因報導分析法證實在PC-3細胞中過度表達p53或以Doxorubicin處理LNCaP細胞都可促進BTG2的表達;而NFκB的活化可抑制LNCaP細胞中BTG2表達並促進細胞生長。p53及NFκB之DNA的結合位置 (-83至-103)互相重疊,故稱此區域為p53-NFκB Response Element Box。另外我們也發現維生素A及Resveratrol刺激可促進BTG2基因表達,其中resveratrol對於BTG2的調控並非透過促進p53或抑制NFκB之途徑,其作用位置在-101至-173之間,可能是透過ERK1/2或Akt的訊息傳遞途徑作用。綜合以上結果,BTG2基因的負向調控參與了前列腺癌的形成,我們可藉p53、NFκB或resveratrol及維生素A調控BTG2基因表達之結果應用於前列腺癌之基因治療上。
B cell translocation gene 2 (BTG2) is a member of anti-proliferative gene family. It regulates G1/S transition of the cell cycle by Rb-dependent and Rb-independent pathways induced by p53 in response to DNA damage. Using RT-PCR to analyze BTG2 gene expression in the prostate carcinoma cells, we found that the BTG2 gene expression is negatively relative to neoplasia of prostate cells. Results from [3H]thymidine incorporation assay demonstrated that overexpression of BTG2 attenuated proliferation of prostate cancer cells, PC-3. Several transcription response elements including p53 and NFκB were found in the promoter region of BTG2 gene. Transient gene transcription assays using the reporter vector containing 5’-flanking region (-1 to -297) of BTG2 gene indicated that BTG2 is a p53-upregulated and NFκB-downregulated gene. Results from this thesis suggested that the p53-NFκB response element box at BTG2 gene (-83 to -103) plays an essential role in BTG2 gene regulation in prostate carcinoma cells. Other studies also showed that Vitamin A and resveratrol were able to induce BTG2 expression in prostate cancer cells. However, resveratrol enhanced BTG2 promoter activity neither by p53-inducing nor NFκB-reducing pathways. The regulatory site of resveratrol was probably at -101 to -173 region of BTG2 gene through Erk1/2 and Akt pathways. In summary, downregulation of BTG2 gene expression takes part in oncogenesis of prostate cancer, suggesting that p53 and NFκB are both significant in BTG2 gene regulation.
序論 …………………………………………………………....……….01
1. Prostate Cancer ………………………………………………..02
2. APRO Gene Family …....…………………………...………….02
3. BTG2 ……………………………………………....…………...09
4. p53 ……………………………………………………………..17
5. NFκB …………………………………………………………..18
6. Resveratrol ...……………………………………………..………22
7. Vitamin A ……………………………………………………24
研究動機與目標……………...………………………...…………..26
實驗材料與方法 ...……………………………………………………27
實驗結果 ………………………………..………………..……………46
1. BTG2表達的降低與前列腺腫瘤形成有關 …………………47
2. 過度表達BTG2基因可抑制前列腺癌細胞生長 ….…………48
3. DNA損害物質可透過p53來促進BTG2的轉錄表達 ……..…48
4. NIK、IκBα對NFκB的調控 ……………………………….…..49
5. 在前列腺癌細胞中NFκB會抑制BTG2的表達 …..……...……50
6. NFκB促進前列腺癌細胞的生長 …………………………….52
7. NFκB對BTG2轉錄調控的位置 …………………..………….53
8. p53對BTG2轉錄調控的位置 ………………………………..54
9. 維生素A及Resveratrol可促進BTG2的表達 ……………….55
10. Resveratrol對於BTG2的調控沒有經p53及NFκB的訊息傳導
路徑 …………………………………………………………56
11. Resveratrol透過ERK1/2或Akt的途徑來調控BTG2 ………57
討論 ………………………………………………………………………59
附錄 ………………………………………………………………………66
1.BTG2 expression is negatively related to prostate neoplasia ……67
2.Overexpression of BTG2 inhibites PC-3 cell growth ……………68
3.Doxorubicin and p53 induce BTG2 expression in prostate cancer cells .........69
4.Effects of IκBα and NIK on NFκB activation in PC-3 and LNCaP cells ………………………………………………………………………70
5.NFκB inhibits BTG2 expression in prostate cancer cells ……………71
6.NFκB induces prostate cell growth ……………………………………73
7.-94 to -102 is the region of NFκB regulation on BTG2 expression …74
8.-83 to -102 is the region of p53 regulation BTG2 on expression …76
9.Vitamin A and resveratrol upragulate BTG2 expression ………77
10.Resveratrol up-regulates BTG2 between -101 to -173 region on BTG2 promoter/enhancer without p53 and NFκB ……………………………78
11.Resveratrol upregulates BTG2 through Akt and ERK1/2 signaling pathways ………………………………………………………………80

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