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

施純如(Chun-Ru Shih)

指導教授 ： 李文森

臺北醫學大學/醫學院/醫學科學研究所/碩士(2002年)

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。

關鍵字

內皮細胞 ； 細胞週期 ； 抗血管新生

並列摘要

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.

並列關鍵字

HUVEC ； cell cycle ； angiogenesis ； SDil-1

參考文獻

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