資料載入中.....
|
請使用永久網址來引用或連結此文件:
http://ir.cmu.edu.tw/ir/handle/310903500/24184
|
| 題名: | Apigenin, 17α-estradiol 和flavone引起人類胃癌細胞株(SC-M1)生長的抑制和調節細胞週期相關基因的表現;Apigenin, 17α-estradiol and flavone induced growth inhibition and modulated cell cycle related gene expression in human stomach adenocarcinoma cell line (SC-M1) |
| 作者: | 王文玲;Wang Wen-Ling |
| 貢獻者: | 中國醫藥學院醫學研究所 |
| 關鍵詞: | 人類胃癌細胞株;apigenin;17α-estradiol;flavone;cell cycle;DD RT-PCR;SC-M1 |
| 日期: | 1991 |
| 上傳時間: | 2009-12-03 09:59:56 (UTC+8) |
| 摘要: | Apigenin、flavone和 17α-estradiol 是屬於植物性荷爾蒙 (phytoestrogens)，廣泛存在於蔬菜及水果中，例如：芹菜、堅果類、茶葉及豆類食物等。此類成分目前已被發現對於癌症有很好治療及預防效果，例如：大腸直腸癌、前列腺癌、血癌和皮膚癌等。本實驗主要利用 apigenin、flavone 和 17α-estradiol 三種成分分別研究它們對人類胃癌細胞株 (SC-M1) 生長、細胞週期、基因表現和乙醯轉化酵素活性的影響。我們利用流式細胞計數儀 (Flow cytometor) 分析細胞存活率、細胞週期及其相關酵素的影響。乙醯轉移酵素受此三種成分的影響是由高壓液相層析儀來分析。實驗結果證明 apigenin、flavone 和 17α-estradiol 均對人類胃癌細胞株有毒殺作用且造成胃癌細胞株的細胞週期停止在G2/M期。在細胞週期素方面，利用流式細胞計數儀和反轉錄酶-聚合酶連鎖反應 (RT-PCR) 結果證明 apigenin 在濃度 60 ìM 抑制 cyclin B1、cyclin D1、CDK1 和 p53 蛋白質及 mRNA的表現，且隨著濃度增加抑制作用更明顯。 Flavone 於 60 μM 抑制 cyclin A和CDK1 蛋白質的表現，在 mRNA 方面則發現抑制 cyclin B1、E 和 p21 mRNA的表現，對於 p53 和 CDK2 mRNA 則是沒有影響。在 60 μM 17α-estradiol同樣發現明顯抑制 cyclin B1 和 CDK1 蛋白質及 cyclin E 和 p53 mRNA 的表現，而 p21 mRNA 的表現則是被促進表現，cyclin B1、D1 和 CDK2 mRNA均不受 17α-estradiol 的影響。 在基因表現方面試利用Differential Display RT-PCR (DD RT-PCR) 的方法鑑定apigenin、flavone和 17α-estradiol對胃癌細胞株 (SC-M1) 基因表現的影響。結果發現17a-estradiol誘導 oncostatin M receptor基因的表現，因此我們再利用 RT-PCR 和流式細胞計數儀再次證明結果與 DD RT-PCR是相符合的。P38 MAPK 為 oncostatin M 蛋白質的其中一個下游訊息傳遞因子，其可調控細胞凋亡和細胞生長，因此本實驗利用 p38 MAPK 抑制劑 SB203580先加入細胞培養液中反應3小時後再加入 60 μM 17α-estradiol培養24小時，利用流式細胞計數儀分析細胞存活率結果發現細胞存活率增加6 %，因此推論17α-estradiol是藉由p38 MAPK 訊息傳遞路徑而導致人類胃癌細胞株產生細胞凋亡。本實驗利用高壓液相層析儀檢測人類胃癌細胞株乙醯轉移酵素活性結果發現apigenin、flavone和17α-estradiol均會抑制乙醯轉移酵素的活性，而apigenin同時也會抑制已醯轉儀酵素mRNA的表現。; Apigenin, flavone and 17α-estradiol are belong to the phytoestrogens and they are present in our common fruits and vegetables such as tea leafs, clear, nuts etc. These compounds have been demonstrated to present cancer therapy and prevention such as colorectal, prostate, leukemia and skin cancers. The purpose of present study is to examine apigenin, flavone and 17α-estradiol whether or not could affect the growth, gene expression, cell cycle, cyclins and N-acetyltransferase (NAT) activity from human stomach cancer cell line (SC-M1). Cell viability, cell cycle and related enzymes (cyclins and cyclin dependent kinases) were examined and determined by flow cytometric assays (FACS). The effects of three compounds on the NAT activity were determined by high performance liquid chromatography (HPLC). The data from experiments indicated that apigenin, flavone and 17α-estradiol induced cytotoxicity, G2/M phase arrest of cell cycle on SC-M1 cells. The cyclins and cyclin dependent kinases examination were performed by FACS and RT-PCR both methods. The results demonstrated that 60 μM apigenin inhibit cyclin B1, CDK1, cyclin E and p53 protein and mRNA expression and this effects is dose dependent; 60 mM flavone inhibit cyclinA and CDK1 protein and cyclin B1, E and P21 mRNA expression but it did not affect CDK2 and P53 MRNA expression; 60 μM 17α-estradiol inhibit cyclin B1 and CDK1 protein expression, it not inhibit cyclin B1, D1 and CDK2 mRNA expression but it inhibit Cyclin E and p53 mRNA expression and promote p21 mRNE expression. The experiments also used differential display RT-PCR (DD RT-PCR) for determining apigenin, flavone and 17α-estradiol affect SC-M1 gene expression. The results demonstrated that 17α-estradiol promote gene expression of oncostatin M receptor from SC-M1 cells. It was also been confirmed by RT-PCR and FACS methods. The P38 MAPK is a down regulation factor for oncostatin M receptor that can regulate cell apoptosis and viability. The inhibitor (SB203580) of P38 MAPK was added to the cells for 3 hours then 60 μM 17α-estradiol was added to the cells the cell viability was increased about 6%. Therefore, 17α-estradiol is through the P38 MAPK pathway to induce cell apoptosis. The data also demonstrated that apigenin, flavone and 17α-estradiol affect NAT activity from SC-M1 cells which was based on the changes of the amounts of N-acetylation of 2-aminofluorene that was determined by HPLC. |
| 顯示於類別: | [醫學研究所] 博碩士論文
|
文件中的檔案:
| 檔案 |
描述 |
大小 | 格式 | 瀏覽次數 |
|---|
| 910704-畢業論文完成版-1.pdf | | 3168Kb | Adobe PDF | 2023 | 檢視/開啟 |
|
在CMUR中所有的資料項目都受到原著作權保護.
|
