氯化两面针碱抑制肿瘤血管生成相关靶标蛋白的初步筛选  

A preliminary screening of molecular targets for anti-tumor angiogenesis effect of nitidine chloride

作　　者：傅宣皓 秦悦[1] 黄燚燚 谢雪平[1] 成晓静[2] 刘华钢[1] 赖泽锋[1] 

机构地区：[1]广西医科大学药学院,南宁530021 [2]广西医科大学生命科学研究院,南宁530021

出　　处：《广西医科大学学报》2017年第5期650-653,共4页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81260657)

摘　　要：目的:初步筛选出氯化两面针碱(NC)抑制肿瘤血管生成的分子靶标。方法:将人脐静脉血管内皮细胞(HUVEC)分为空白对照组及不同浓度(30μmol/L和60μmol/L)NC的实验组,采用细胞划痕实验检测经NC干预24h和48h后HUVEC的迁移能力;western blotting法检测NC干预后HUVEC中血管内皮生长因子(VEGF)的蛋白表达水平。以计算机辅助药物设计软件SYBYL 2.0对NC与肿瘤血管生成相关信号通路受体蛋白进行分子对接,以Pymol软件绘制三维结构图并分析NC与受体蛋白的相互作用。结果:与空白对照组比较,30μmol/L和60μmol/L NC实验组细胞迁移率显著降低(P<0.001),各实验组的VEGF蛋白表达水平均显著下调(P<0.01)。分子对接结果表明,在肿瘤血管生成信号通路中NC与磷脂酰肌醇3-激酶(PI3K)和非受体酪氨酸激酶c-Src主要通过氢键和疏水场相互作用,亲和力较强。结论:NC能够下调VEGF的表达来抑制HUVEC的迁移,进而抑制肿瘤血管生成,PI3K和c-Src是NC抑制肿瘤血管生成的潜在靶标。Objective：To preliminarily screen the molecular targets of nitidine chloride （NC） relevant to its anti-tumor angiogenesis effect. Methods： Human umbilical vein endothelial cells （HUVECs） were divided into a control group and two different doses （30 μmol/L and 60 μmol/L） NC groups. The migration ability of HUVECs was detected by wound healing assay after exposure to NC for 24 h and 48 h. The protein expression of vascular endothelial growth factor （VEGF） in HUVECs was determined by western blotting after NC treatment. The computer-aided drug design （CADD） software SYBYL 2.0 was employed to conduct the molecular docking between NC and the receptors from the tumor angiogenesis signals, and the Pymol software was used to draw the 3-dimension structures of the docking results. The interaction between NC and the receptors was analyzed. Results： Compared with the control group, the cell migration rate in 30 /.tmol/L and 60 μmol/L NC groups was significantly decreased （P〈0. 001）. The protein expression of VEGF in NC groups was down-regulated （ P 〈0. 01）. The molecular docking results indicated that NC had strong affinity to both phosphatidylinositol 3-kinase （PI3K） and non-receptor tyrosine kinase （c-Src） and their interactions were dominantly hydrogen bonds and hydrophobic interaction. Conclusion： NC inhibited the migration of HUVECs via suppressing the expression of VEGF. PI3K and c-Src might be potential molecular targets relevant to anti-tumor and anti-angiogenesis effects of NC.

关 键 词：氯化两面针碱 血管生成 分子对接 靶标 PI3K C-SRC 

分 类 号：R285.5[医药卫生—中药学] R966[医药卫生—中医学]