交叉反应物质197转运大分子物质通过血肿瘤屏障的效果和机制  

Effects and mechanisms of CRM197 on delivering macromolecular substance across the blood-tumor barrier

作　　者：王萍[1] 刘云会[2] 薛一雪[1] 

机构地区：[1]中国医科大学基础医学院神经生物学教研室,辽宁沈阳110001 [2]中国医科大学附属盛京医院,辽宁沈阳110004

出　　处：《中国药理学通报》2011年第7期907-910,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81072056;30973079);教育部博士点学科专项科研基金资助项目(No20092104110015;20102104110009);沈阳科学技术计划资助项目(NoF10-205-1-37;F10-205-1-22)

摘　　要：目的研究白喉毒素的无毒突变体交叉反应物质197(cross-reacting material 197,CRM197)转运大分子物质通过血肿瘤屏障的效果和机制。方法制备CRM197-HRP偶联物,建立体外血肿瘤屏障模型,给予CRM197-HRP作用2 h后,通过TMB显色法检测transwell下室中HRP活性的变化;给予CRM197作用1 h后,采用免疫组化法检测体外血肿瘤屏障内皮细胞中p-Akt的表达变化;Western blot法检测p-Akt、p-FOXO1A和质膜微囊蛋白caveolin-1的表达变化。结果和对照组相比,CRM197-HRP能有效通过血肿瘤屏障,并且其转运水平呈时间依赖性增加;p-Akt主要分布在内皮细胞的胞质和胞核中,CRM197作用1 h后p-Akt的表达水平下降;同时伴有内皮细胞中p-FOXO1A表达下降和质膜微囊蛋白caveolin-1的表达上调。结论 CRM197可能通过抑制内皮细胞中p-Akt的表达,减少转录因子FOXO1A的磷酸化水平,进一步上调质膜微囊蛋白caveolin-1的表达,增加血肿瘤屏障的通透性。Aim To investigate the effects and mechanisms of CRM197, the non-toxic mutant of diphtheria toxin, on delivering the macromolecular substance across the blood-tumor barrier（BTB）. Methods The CRM197-HRP conjugate was prepared and the BTB model was established in vitro. After treatment of CRM197-HRP for 2 h,TMB coloration was used to detect the HRP activity in the basal chamber of transwell. Immunohistochemical analysis was used to detect the expression of p-Akt in the endothelial cells of BTB in vitro. Western blot assay was used to detect the expressing levels of p-Akt, p-FOXO1A and caveolin-1. Results Compared with control group, CRM197-HRP could get through the BTB effectively, and this transcytosis increased in a time-dependent manner. Phospho-Akt located mainly in the cytoplasm and nucleus of endothelial cells. After treatment of CRM197 for 1 h,the expressing level of p-Akt reduced significantly. This was accompanied by the decrease of p-FOXO1A and the up-regulation of caveolin-I in endothelial cells. Conclusion CRM197 might decrease the phosphoryl- ation level of transcription factor FOXO1A by inhibiting the expression of p-Akt in endothelial cells, and then up-regulate the express of caveolin-1 and increase the BTB permeability.

关 键 词：CRM197 血肿瘤屏障 P-AKT p-FOXO1A CAVEOLIN-1 通透性 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]