人类非小细胞肺癌细胞对小鼠体外血脑屏障模型功能的影响

发布时间：2018-04-30 17:10

本文选题：血脑屏障模型 + 非小细胞肺癌　；参考：《神经解剖学杂志》2017年02期

【摘要】：目的:建立体外血脑屏障模型(blood-brain barrier model,BBBM),观察非小细胞肺癌(non-small cell lung cancer,NSCLC)A549细胞对BBBM功能的影响。方法:早上将周细胞(pericytes,PC)接种到Transwell膜的外侧面,星形胶质细胞(astrocytes,AS)接种到12孔板内,8 h后将Transwell置入12孔板内,再将小鼠脑微血管内皮细胞(brain microvascular endothelial cells,BMEC)接种到transwell膜的内侧面,建立血脑屏障模型。模型建立后每天进行试漏试验及跨内皮电阻(transendothelial electrical resistance,TEER)测定,鉴定是否成模。成模后分为对照组与实验组,将A549细胞接种到实验组BBBM内室,共培养体系置于培养箱,于24 h、48 h、72 h后进行电阻测定及辣根过氧化物酶渗透试验,以评估BBBM的功能。分别于渗透试验进行至2 h、6 h时取样,根据样本中辣根过氧化物酶浓度计算表观渗透系数(apparent permeability coef錞cient,Papp)。结果:(1)部分血脑屏障模型在建立后3 d成模,4 d可全部成模。(2)A549细胞加入BBBM内室共培养。(a)实验组TEER值与对照组比较:1 d后无显著变化(P0.05);2 d后降低出现统计学差异(P0.05);3 d后降低具有统计学差异(P0.01);(b)渗透试验与对照组比较:1 d后2 h及6 h渗透试验实验组Papp均降低,但无统计学差异(P0.05);2 d后实验组Papp升高,2 h渗透试验差异具有统计学意义(P0.05),6 h渗透试验差异无统计学意义(P0.05);3 d后实验组Papp升高,2 h及6 h渗透试验均具有显著统计学差异(P0.01)。结论:小鼠体外血脑屏障模型的功能可被人类非小细胞肺癌细胞影响。体外共培养体系可用于研究肺癌脑转移过程中肿瘤细胞与脑微血管内皮细胞间相互作用的分子机制。
[Abstract]:Aim: to establish a blood-brain barrier model in vitro to observe the effect of non-small cell lung cancer on the function of BBBM in NSCLC A549 cells. Methods: in the morning, the pericytes were inoculated into the outer side of the Transwell membrane, the astrocytes were inoculated into the 12-hole plate for 8 hours, the Transwell was placed into the 12-hole plate, and then the mouse brain microvascular endothelial cells were inoculated into the inner side of the transwell membrane. Blood brain barrier model was established. After the establishment of the model, the leakage test and transendothelial electrical resistance test were carried out every day to determine whether the model was formed. The model was divided into control group and experimental group. A549 cells were inoculated into the BBBM chamber of experimental group. The co-culture system was placed in incubator. After 24 h, 48 h and 72 h, resistance measurement and horseradish peroxidase permeation test were performed to evaluate the function of BBBM. The peroxidase concentration of horseradish peroxidase was calculated according to the apparent osmotic coefficient of permeability coef. Results part of the blood-brain barrier model was established 3 days after the establishment of the model and 4 days after the establishment of the model, all of the cells were added into the BBBM chamber co-culture. The TEER value of the experimental group was no significant change compared with that of the control group after 1 day. P0.05A549 cells decreased after 2 days. There was a statistical difference between the experimental group and the control group. Compared with the control group, the Papp of the experimental group at 2 h and 6 h after 1 day was significantly lower than that of the control group. However, there was no statistical difference between the experimental group and the experimental group after 2 days after P0.05, there was no significant difference between the experimental group and the experimental group. There was no significant difference in the permeability test between the experimental group and the experimental group. There was no significant difference between the experimental group and the experimental group. After 3 days, the Papp in the experimental group was increased for 2 h and 6 h, respectively. There was significant difference between the two groups (P0.01) and the experimental group (P0.01). Conclusion: the function of mouse blood-brain barrier model can be affected by human non-small cell lung cancer cells in vitro. In vitro coculture system can be used to study the molecular mechanism of the interaction between tumor cells and microvascular endothelial cells during brain metastasis of lung cancer.
【作者单位】：昆明医科大学第一附属医院神经外科;红河州第一人民医院;
【基金】：云南省科技厅-昆明医科大学应用基础研究联合专项资金项目(2015FB033)
【分类号】：R734.2;R-332

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