肿瘤源性血管内皮细胞的培养及靶向CD105能力测定  

Culture of tumor-derived vascular endothelial cells and identification of their molecular targeting ability

作　　者：杨华[1] 龚明福[2] 徐建众 邹利光[2] 张松[2] YANG Hua1,GONG Mingfu2,XU Jianzhong1,ZOU Liguang2,ZHANG Song2(1. Department of Radiology, Chongqing Traditional Chinese Medicine Hospital, Chongqing 400021, China ; 2. Department of Radiology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, Chin)

机构地区：[1]重庆市中医院放射科,重庆400021 [2]第三军医大学新桥医院放射科,重庆400037

出　　处：《西部医学》2018年第5期636-640,共5页Medical Journal of West China

基　　金：国家自然科学基金(81071197;81501521);重庆市前沿与应用基础研究项目(cstc2015jcyjA1338)

摘　　要：目的探讨肿瘤源性血管内皮细胞(Td-VECs)的培养方法,并测定CD105表达及靶向能力。方法采用酶消化法分离、培养人脐静脉内皮细胞(HUVECs),CD34免疫荧光染色鉴定;Millicell小室共培养VECs和乳腺癌细胞以获取肿瘤血管的内皮细胞(Td-VECs),采用聚合酶链反应、CD105免疫荧光及靶向CD105纳米粒标记检测Td-VECs细胞特性、分子表达及靶向结合能力。结果成功分离HUVECs,诱导后VECs的Tem1和Tem8表达量明显增高,免疫荧光染色显示VECs呈CD105阳性,诱导后的VECs荧光强度强于未诱导VECs;在铁浓度为0、0.5、1、2、5、10ug/mL时,共同孵育24h,诱导后和未诱导的VECs的标记率分别为0%、(25.35±4.23)%、(58.07±4.12)%、(86.68±3.42)%、100%、100%和0%、(10.58±2.62)%、(16.31±4.32)%、(46.33±3.42)%、(77.62±4.13)%、100%。结论肿瘤细胞共培养法能够诱导VECs向Td-VECs分化,是获取基于CD105分子靶向研究Td-VECs的可靠方法,为靶向抗肿瘤血管生成提供了细胞学基础。Objective To investigate the culture of tumor-derived vascular endothelial cells（Td-VECs）and identify their CD105 expression and targeting ability.Methods Enzyme digestion was used to isolate human umbilical vein endothelial cells（HUVECs）.CD34 immunofluorescence staining was used to identify HUVECs.In order to acquire the characteristics of Td-VECs,VECs were incubated with breast cancer cells in Millicell cell culture.The cell characteristics,molecular expression and targeted binding ability of Td-VECs were detected by polymerase chain reaction（PCR）,CD105 immunofluorescence and CD105-targeted nanoparticles.Results HUVECs were successfully isolated.The expression of Tem1 and Tem8 in VECs was significantly increased after induction with breast cancer cells.Incubated VECs were positive for CD105 immunofluorescence staining,and the fluorescence intensity of induced VECs was stronger than that of non-induced VECs.After co-incubation for 24 hours,at iron concentrations of 0,0.5,1,2,5,10 ug/mL,the labeling ratios were,respectively,0%,（25.35±4.23）%,（58.07±4.12）%,（86.68±3.42）%,100% and 100%for induced VECs,and 0%,（10.58±2.62）%,（16.31±4.32）%,（46.33±3.42）%,（77.62±4.13）%,100% for non-induced VECs.Conclusion Tumor cell co-culture method can induce VECs to differentiate into Td-VECs and is a reliable method to obtain Td-VECs for CD105 molecular targeting studies.

关 键 词：血管内皮细胞 肿瘤 培养 鉴定 CD105 

分 类 号：R393[医药卫生—基础医学] R445