肺癌脑转移型A549/GFP-2细胞的筛选及其条件液对脑微血管内皮细胞的作用

The Selection of A549/GFP-2 of Brain Metastases From Lung and the Effect of Condition Medium on Brain Microvascular Endothelial Cells

目的:建立肺癌脑转移模型,筛选脑转移倾向细胞A549/GFP-2,探讨A549和A549/GFP-2条件液对脑微血管内皮细胞的作用,揭示肺癌脑转移的机制。方法:利用胸腔原位注射法筛选出A549脑转移细胞亚型A549/GFP-2,原代培养大鼠脑微血管内皮细胞,观察A549和A549/GFP-2细胞条件液对脑微血管内皮细胞增殖的影响和细胞内HIF-1α和VEGF表达的改变。结果:胸腔内原位种植较好地反应了临床肺癌脑转移的过程。不同浓度的A549和A549/GFP-2细胞条件液对脑微血管内皮细胞增殖的影响不同,低浓度(<30%)对脑微血管内皮细胞有促进的作用;高浓度(>60%)对脑微血管内皮细胞的增殖有不同程度的抑制作用,且有随浓度增加抑制作用增强的趋势。A549和A549/GFP-2细胞条件液能提高脑微血管内皮细胞内HIF-1α和VEGF的表达。结论:胸腔内原位种植是建立肺癌脑转移的稳定模型。肺癌脑转移与肺癌细胞在生长过程中分泌的HIF-1α和VEGF等细胞因子破坏了脑微血管内皮细胞的结构有关。

Objective： The model of brain metastases was established from lung, and the effect of A549 and A549/GFP-2 condition medium was explored on Cerebral Microvascular Endothelial Cells （CMEC）. So the mechanism of brain metastases from lung was indicted. Methods： A549/GFP-2 cells have been selected from A549 cells by orthotopic implantation in chest, and primary cultured rat cerebral microvascular endothelial cell. The proliferation have been observed in CMEC and the expression of HIF-lalpha and VEGF have been evalfiated in CMEC. Results： The orthotopic implantation of A549 was the bettest way for brain metastases of lung cancer in chest. Low concentration （〈30%） could promote proliferation of CMEC, but high concentrations （〉60%） could inhibit proliferation of CMEC. And the effect of inhibition enhanced with increasing concentration. The conditioned medium of A549 and A549/GFP-2 cells could increase the expression of HIF-lalpha and VEGF in CMEC. Conclusion： Orthotopic implantation in chest is the stable model for brain metastases from lung. The brain metastasis from lung cancer relates to destruction of structure of CMEC.