臺灣博碩士論文加值系統

中 文 摘 要
苯駢(a)芘是一種廣泛存在我們生活環境中的污染物,在許多基因毒性與實驗動物的研究中顯示,它具有細胞致癌能力,但其致癌的機制至今仍不是完全清楚.因此,我們研究苯駢(a)芘對細胞質(蛋白激C),細胞核(原致癌基因：c-jun，c-fos )及細胞轉形能力（細胞生長複製時間、細胞生長飽和度及細胞轉形分析）的影響,進而推測致癌化過程中可能的分子機制.
在細胞生長複製時間與飽和密度實驗的結果中發現,當處理苯駢(a)芘低劑量(1-5 μM)時,就會使細胞複製時間縮短,細胞飽和密度增加,將苯駢(a)芘與一個強的促癌劑12-o-tetradecanoyl phorbol-13-acetate(TPA)一起作用細胞時則複製時間將明顯縮短,飽和密度明顯劇增.進行細胞轉形分析時,發現單獨以苯駢(a)芘處理細胞時,雖然苯駢(a)芘可以致細胞轉形產生群落但不較單獨處理N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)時的轉形能力強,卻比單獨處理TPA時轉形能力強,可是如果將苯駢(a)芘加上TPA一起作用,則致轉形能力與陽性對照組(MNNG+TPA) 相接近,這意味著苯駢(a)芘在細胞轉形能力上似乎是個啟始劑角色. 蛋白激C在細胞訊息傳遞上佔有樞杻的地位,因此我們進一步分析苯駢(a)芘對蛋白激C的影響,從西方墨點法中發現苯駢(a)芘在劑量與時間變化上對於蛋白激C的α及δisoform的表現均無影響 ,已知c-Fos與c-Jun這兩種蛋白可以促進細胞週期的進行,在北方墨點法中發現苯駢(a)芘在劑量與時間變化上對於原致癌基因c-fos與c-jun的表現沒有影響 ,溶劑DMSO會影響c-jun基因的表現.綜合上述的結果,我們發現苯駢(a)芘在低劑量時就可以改變細胞的某些生長特性,於細胞訊息傳遞上不是經由活化蛋白激C及誘發原致癌基因c-fos與c-jun的這條路徑,在致癌性上苯駢(a)芘雖然可以引起細胞轉形,但是它似乎只扮演著一個啟始劑的角色。

Abstract
Benzo(a)pyrene( B(a)P ) is a kind of environmental pollutants that commonly exists in our circumstance. It has been shown that B(a)P is carcinogenic in many researches of genotoxicities and experimental animals , but the underlying mechanism is still not clear. To surmise the possible molecular mechanisms, we studied the effects of B(a)P on the molecules in cytoplasm ( protein kinase C ) and nucleus ( proto-oncogene : c-jun , c-fos ) , individually , and B(a)P's transformation ability by performing doubling time , saturation density , transformation assays. From the doubling time and saturation density tests , we found that the cells doubling time was shortened and its saturation density was increased when the cells were treated with B(a)P (1-5 uM). Combined treatment of both B(a)P and the well known tumor promotor 12-o-tetradecanoyl phorbol 13-acetate (TPA) enhanced these observed phenomena. In cell transformation test , we observed that treatment of B(a)P alone transformed cells to foci formation. Its transformation ability was not as good as the well-known carrcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) , but better than TPA. If B(a)P was combined with TPA , the transformation ability was similar to positive control (MNNG+TPA). These results suggested that B(a)P could play a role as an initiator but not promotor during cell transformation. Protein kinase C (PKC) is pivotal in intracellular signal transduction, Therefore we analyzed B(a)P's effect on PKC translocation. From western blot results , B(a)P did not affect PKC-αandδisoform translocation. As we known , c-Jun and c-Fos are proteins that accelerate the processing of cell cycle. From northern blot analysis , B(a)P did not have significant effect on c-jun and c-fos expression. In summary , our data suggest that B(a)P may alter some events related to cell growth but not via PKC signal transduction and c-jun and c-fos expression. During carcinogenesis , B(a)P causes cell transformation and may play a role as an initiator.

目 錄
中文摘要------------------- 1
英文摘要------------------- 3
緒言----------------------- 5
材料與方法----------------- 9
結果----------------------- 19
討論----------------------- 28
參考文獻------------------- 32
表格與圖------------------- 41
致謝------------------------73

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