新型蛋白酶体抑制剂YSY-01A对肿瘤细胞促血管生成作用的抑制及其机制初探  

Inhibition Effect and Mechanism of YSY-01A,a Novel Proteasome Inhibitor,on Tumor-induced Angiogenesis

作　　者：刘敬弢[1] 袁霞[1] 徐波[1] 冉福香[1] 楚明明[1] 贾璇[1] 陈溢欣[1] 王哲[1] 李润涛[1] 崔景荣[1] 

机构地区：[1]北京大学药学院,天然药物与仿生药物国家重点实验室,北京100191

出　　处：《生物化学与生物物理进展》2013年第8期748-756,共9页Progress In Biochemistry and Biophysics

基　　金：国家“十一五”科技重大专项:北京大学综合性创新药物研究开发技术大平台(2009ZX09301-010)~~

摘　　要：YSY-01A是一种新型蛋白酶体抑制剂,前期研究已经证实其对肿瘤细胞的增殖有抑制作用.但是它对肿瘤血管生成是否有影响尚不明确.本研究旨在探明YSY-01A阻碍肿瘤细胞促进血管生成的作用及机制.我们首先将磺酰罗丹明B(sulforhodamine B,SRB)法与细胞共培养(Transwell)模型相结合,探讨YSY-01A抑制人结肠癌细胞(HT-29 cells)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)的增殖促进作用;运用高内涵筛选(high content screening,HCS)法研究YSY-01A对HT-29细胞中NF-κB核转位的影响;利用Western blot法检测YSY-01A对HT-29细胞中缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)及血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达调控.为了观察YSY-01A对HUVEC增殖和运动有无直接抑制作用,我们采用SRB法观察YSY-01A对HUVEC的增殖抑制作用;运用HCS法分别考察YSY-01A对HUVEC的运动抑制和细胞毒作用.结果证实,YSY-01A可以阻碍HT-29细胞对HUVEC的增殖促进作用并具有浓度依赖性.YSY-01A还可抑制HT-29细胞中NF-κB的核转位,下调HIF-1α及VEGF的表达.进一步研究证实,YSY-01A能够浓度依赖地抑制HUVECs的增殖和运动,而不伴有明显的细胞毒作用.上述结果表明,YSY-01A可以通过抑制蛋白酶体活性下调肿瘤细胞中促血管生成因子的表达,进而在血管内皮细胞中发挥抗血管生成作用.Compound YSY-01A is a recently synthesized proteasome inhibitor. It has been proved for potent growth-inhibitory effect on tumor cells in previous studies. However, the effect of YSY-01A on tumor angiogenesis remains unclear. Our research aims to reveal the inhibition effect and mechanism of YSY-01A on tumor-induced angiogenesis. Firstly, we combined the Sulforhodamine B （SRB） assay and Transwell co-culture model to observe the inhibition of YSY-01A on human umbilical vein endothelial cells （HUVECs） proliferation induced by tumor cells （HT-29 cells）. In succession, high content screening （HCS） assay was used to investigate effect of YSY-01A on NF-KB nuclear translocation in HT-29 cells. Finally, Western blot was used to measure the expression of hypoxia-inducible factor-loL （HIF-1α） and vascular endothelial growth factor （VEGF） in HT-29 cells inhibited by YSY-01A. To further determine mechanism of inhibition, SRB and HCS methods were used to investigate the effect of YSY-01A against HUVECs proliferation and motility, respectively. The results showed that YSY-01A could prohibit HUVECs proliferation induced by HT-29 cells in a concentration-dependent manner. Furthermore, YSY-01A significantly inhibited NF-KB nuclear translocation and reduced the expression of HIF-1α and VEGF in HT-29 cells. Further investigation revealed concentration-dependent suppress of YSY-01A on HUVECs proliferation and motility without obvious cytotoxic effect. In conclusion, through proteasome inhibition, YSY-01A could down-regulate pro-angiogenesis factors expression in tumor cells and exhibit remarkable anti-angiogenesis activity on vascular endothelial cells.

关 键 词：蛋白酶体抑制剂 血管生成 人脐静脉内皮细胞 HT-29细胞 高内涵筛选 YSY-01A 

分 类 号：R979.1[医药卫生—药品] R965.11[医药卫生—药学]