Anip973细胞上清液增加脐静脉内皮细胞的生物活性  

The Supernatant Obtained from Cultured Anip973 Cells Enhances the Biological Activities of HUVEC

作　　者：张翠翠[1] 李月雅[1] 王晶[1] 李凯[1] 

机构地区：[1]天津医科大学肿瘤医院肺部肿瘤内科,国家肿瘤临床医学研究中心,天津市"肿瘤防治"重点实验室,天津市肺癌诊治中心,天津300060

出　　处：《中国肺癌杂志》2015年第11期668-673,共6页Chinese Journal of Lung Cancer

基　　金：天津市科委课题(No.11JCYBJC11300);国家自然科学基金(No.81372517);国家科技重大专项(No.2013ZX09303001);院级课题(No.1317);CSCO-先声抗肿瘤血管靶向治疗科研基金(No.Y-S2014-011)资助~~

摘　　要：背景与目的肿瘤微血管内皮细胞与正常组织来源的微血管内皮细胞相比,具有对生长因子反应性高、粘附因子表达高等特点,造成肿瘤血管通透性高且新生旺盛,导致肿瘤的快速生长和转移。因此了解肿瘤微环境中内皮细胞发生、形态及功能上的异质性特点,对进行合理的、个性化的、以血管内皮细胞为靶点的抗血管新生治疗有一定的指导作用。本实验旨在研究高转移肺腺癌Anip973细胞培养上清液对人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)表面标记及其生物学行为的影响。方法以含有不同浓度Anip973细胞上清液的培养基(RPMI-1640+10%胎牛血清)培养的HUVEC为实验组、单纯培养基培养的HUVEC为对照组,以CCK-8检测各组细胞增殖率、基质胶成管实验检测成管能力、Transwell检测迁移能力、流式细胞术检测细胞表面标记CD105、CD31以及凋亡标记Annexin V的表达,并分析细胞表面标记的表达与生物学行为之间的关系。结果与对照组相比,经Anip973细胞上清液培养的HUVEC增殖更多、且在浓度为250μL/m L作用24 h最明显(P=0.002);成管及迁移能力亦均较对照组增强,分别在浓度为125μL/m L(P<0.001)、250μL/m L(P=0.002)时达最强;细胞表面标记CD105表达阳性率升高、浓度为250μL/m L(P=0.028)最明显;CD31表达阳性率呈浓度依赖性升高;凋亡细胞比例降低。相关性分析显示:CD105的表达阳性率与细胞增殖及迁移能力呈正相关关系、CD31的表达与生物学行为并无明显相关性。结论 Anip973细胞上清液能促进HUVEC细胞增殖、迁移、血管形成,CD105也随之变化、并可在一定程度上反应其生物学行为。Background and objective Unlike normal tissue-derived microvascular endothelial cells, tumor microvessel endothelial cells are highly reactive to growth factors and exhibit more adhesion molecules. Thus, vascular tumors are highly permeable and grow vigorously; this occurrence results in rapid growth and metastasis cancer cells. Therefore, understanding the characteristics of endothelial cells in the tumor microenvironment guides anti-angiogenic therapy. To this end, we explore the effect of the supernatant obtained from cultured Anip973 cells（high-metastatic human lung adenocarcinoma cells） on the biological behavior and on the cell surface markers of the human umbilical vein endothelial cell（HUVEC）. Methods The HUVEC that was cultured in a medium（RPMI-1640 ＋ 10% fetal bovine serum） containing various concentrations of Anip973 supernatants was categorized into experimental groups. The HUVEC cultured in a medium without Anip973 supernatants served as the control group. Proliferation was determined with CCK-8; blood vessel formation was investigated with three-dimensional culture techniques in vitro; and HUVEC migration was observed via transwell assay. At the same time, the expressions of CD105, CD31, and the apoptotic marker of Annexin V were detected through flow cytometry for analyzing therelationship between the expression of cell surface markers and biological behavior. Results Following incubation with the supernatant obtained from cultured Anip973 cells, HUVEC proliferated more than the control group did, and the proliferation rate was maximized when incubated in a supernatant concentration of 250 μL/m L for 24 h（P=0.002）. In addition, the experimental groups exhibited varying degrees of migration and forms of vascular lumen sample structure, especially at supernatant concentrations of 125 μL/m L（P0.001） and 250 μL/m L（P=0.002）, respectively. CD105 expression was optimized at 250 μL/m L（P=0.028）, and CD31 expression also increased with an increase in concentration. However

关 键 词：ANIP973 HUVEC 细胞增殖 迁移 成管 CD105 CD31 ANNEXIN V 

分 类 号：R734.2[医药卫生—肿瘤]