CXCR4/SDF-1轴调节人肺腺癌PC-9细胞对Bends细胞血脑屏障模型功能的影响  

CXCR4/SDF-1 axis regulates the effect of human lung adenocarcinoma PC-9 cells on function of in vitro blood-brain barrier model formed by Bends cells

作　　者：李鸿茹[1,2] 涂洵崴 陈正伟[1,2] 陈愉生[1,2] 韩莉莉[3] LI Hongru;TU Xunwei;CHEN Zhengwei;CHEN Yusheng;HAN Lili(Department of Respiratory and Critical Medicine,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou 350001,Fujian,China;Fujian Institute of Respiratory Diseases,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou 350001,Fujian,China;Fujian Key Laboratory of Cardiovascular Diseases,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou 350001,Fujian,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院呼吸与危重症医学科,福建福州350001 [2]福建医科大学省立临床医学院福建省立医院福建省呼吸病研究所,福建福州350001 [3]福建医科大学省立临床医学院福建省立医院福建省心血管病重点实验室,福建福州350001

出　　处：《中国肿瘤生物治疗杂志》2020年第5期528-533,共6页Chinese Journal of Cancer Biotherapy

基　　金：福建省卫生健康委中青年骨干人才培养项目资助(No.2019148);福建省立医院高水平医院建设科研联合基金资助项目(No.2018GSP008);福建医科大学启航基金资助项目(No.2018QH1130)。

摘　　要：目的:通过建立体外血脑屏障(blood brain barrier,BBB)模型,探讨人肺腺癌PC-9细胞在CXCR4/SDF-1轴作用下对BBB紧密连接蛋白的影响。方法:利用永生化的小鼠脑微血管内皮细胞Bends进行单层培养,建立体外BBB模型;通过跨内皮细胞电阻(transendothelial electrical resistance,TEER)测定及荧光素钠通透性实验判定体外BBB模型的功能状态以及观察PC-9细胞对体外BBB模型功能的影响。Western blotting检测PC-9细胞在CXCR4抑制剂AMD3100、SDF-1单独或联合(1μg/ml AMD3100,100 ng/ml SDF-1,AMD3100+SDF-1)作用下对BBB模型功能和内皮细胞紧密连接蛋白表达的影响,Transwell迁移实验检测CXCR4/SDF-1轴对PC-9细胞跨BBB模型细胞层迁移能力的影响。结果:Bends细胞单层培养可形成紧密连接的"屏障"并产生较高的TEER,第96 h达到(182.13±5.19)Ω·cm^2;同时行荧光色钠通透性实验结果显示,BBB具有良好屏障性能,其通透率低于空白对照组(P<0.05)。PC-9细胞作用后,BBB模型TEER逐渐降低,第24 h降至(46.7±4.35)Ω·cm^2;同时BBB通透率较作用前显著提高(P<0.05)。PC-9细胞在AMD3100作用下能够上调内皮细胞紧密连接蛋白的表达(P<0.05);AMD3100处理组的PC-9细胞穿过BBB的细胞数较空白组明显减少[(43±2)vs(81±2)个,P<0.05]。结论:AMD3100能够减弱PC-9细胞对Bends细胞建立的体外BBB模型紧密连接的破坏能力。Objective: To investigate the influences of human lung adenocarcinoma PC-9 cells on tight junction proteins of blood brain barrier(BBB) under CXCR4/SDF-1 axis by establishing a model of BBB in vitro. Methods: The immortalized mouse brain microvascular endothelial Bends cells were used to establish a model of BBB in vitro by monolayer culture;Subsequently, transendothelial electric resistance(TEER) and fluorescein sodium permeability experiment were used to detect the function of in vitro BBB model and observe the effect of PC-9 cells on the function of BBB model, respectively. Western blotting was used to detect the effect of PC-9 cells on function of BBB model and expressions of endothelial tight junction proteins under the treatment of single or combined AMD3100 and SDF-1(1 μg/ml AMD3100,100 ng/ml SDF-1, AMD3100+SDF-1). Transwell assay was used to detect the influence of CXCR4/SDF-1 axis on the ability of PC-9 cells transmigrating the cell layer of BBB model. Results: Monolayer culture of Bends cells can form tightly connected BBB with high TEER, which reached(182.13±5.19) Ω.cm^2 at the 96 h;in the meanwhile, fluorescein sodium permeability experiment showed that BBB had significantly lower permeability than that of control group([40.31±2.43]% vs [150.10±3.17]%, P<0.05). The TEER of BBB decreased to(46.7±4.35) Ω·cm^2 after coculture with PC-9 cells for 24 h, and at the same time the sodium fluorescein permeability of BBB significantly increased than that of pre-treatment([136.32±4.93]% vs [50.24±6.21]%, P<0.05).PC-9 cells up-regulated the expressions of tight junction proteins of Bends cells under the treatment of AMD3100(P<0.05). The number of PC-9 cells transmigrating the BBB in AMD3100 treatment group was significantly lower than that of CON group(43±2 vs 81±2,P<0.05). Conclusion: AMD3100 can reduce the ability of PC-9 cells destroying the tight junction of the BBB model established in vitro by Bends cells.

关 键 词：肺腺癌 PC-9细胞 血脑屏障 Bends细胞 紧密连接 跨内皮细胞电阻 迁移 CXCR4/SDF-1轴 

分 类 号：R734.2[医药卫生—肿瘤] R730.2[医药卫生—临床医学]