5,5-Diphenyl-2-thiohydantoin (SDil-1)對於人類血管內皮細胞的生 長抑制作用
The anti-proliferation effect of 5,5-Diphenyl-2-thiohydantoin ( SDil-1 ) in human vascular endothelial cells
中文摘要
本篇論文的主旨,在研究化學合成物質SDil-1
(5,5-Diphenyl-2-thiohydantoin) 對於人類臍帶靜脈內皮細胞 HUVEC 在 體外生長的影響, 並探討其中作用的機制。 我們的初步實驗結果發現 SDil-1 在不造成細胞死亡的濃度下, 能對 HUVEC 產生生長抑制的作用。
3H-Thymidine incorporation 的實驗結果顯示, SDil-1 會抑制內皮細胞 DNA 的合成作用。 利用流式細胞儀分析細胞週期,發現 SDil-1 增加了細胞停 滯在G0/G1 phase 的比例。西方墨點法(Western blot)的實驗結果, 觀 察到SDil-1 處理 21 小時的細胞,其和細胞週期停滯有關的蛋白 p21, 表現 量高於對照組, 同時 cdk4、 cyclin D3、cyclin A 蛋白的表現量有顯著的降 低, p53、p27、cdk2、cyclin D1, 以及 cyclin E 蛋白的表現則無明顯變 化。 免疫沉澱(immunoprecipitation)與蛋白激酶活性測定的實驗結果, 發 現以SDil-1 處理後,能增加 p21 與 cdk2 和 cdk4 的結合量, 並降低 cdk2 的激酶活性。 根據實驗結果, 我們認為 SDil-1 會干擾內皮細胞的細胞週期進 行而減少細胞的增生, 其作用主要是透過抑制 cdk 活性的途徑, 因此, SDil-1 或許具有潛力, 能成為抗血管新生的藥物。
英文摘要
The aim of this study is to examine the antiproliferation effect of SDil-1
(5,5-Diphenyl-2-thiohydantoin)in human umbilical vein endothelial cells
(HUVEC)and its possible underlying mechanism. Our data demonstrated that SDil-1 caused an inhibition in HUVEC proliferation. The result of 3H-Thymidine incorporation showed that SDil-1 decreased DNA synthesis in HUVEC. Flow cytometric analysis demonstrated that treatment of HUVEC with SDil-1 arrested the cell at the G0/G1 phase of the cell cycle. Western blot analysis showed that treatment of HUVEC with SDil-1 for 21h increased the levels of p21 protein but decreased the level of cdk4, cyclin D3, and cyclin A proteins. The expression of p53、p27, cdk2, cyclinD1 and cyclin E levels did not significantly change as compared with untreated cells. Furthermore, immunoprecipitation study showed that the cdk4-cyclin-linked p21 and the cdk2-cyclin-linked p21 proteins were increased after SDil-1 treatment.
Using kinase assay method to measure the kinase activity, we demonstrated that the
cdk2 activity was decreased in the SDil-1 treated HUVEC. Thus, our results suggest that SDil-1 can interrupt the cell cycle progression and proliferation of human
endothelial cells by inhibiting the cyclin-dependent kinase activity. The findings from the present study suggest that SDil-1 might have the potential to inhibit the
occurrence of angiogenesis.