摘要
目的小细胞肺癌易发生脑转移,但其机制尚不完全清楚,本文探讨小细胞肺癌细胞对人脑微血管内皮细胞单层细胞间紧密连接开放的作用。方法应用免疫荧光技术观察人脑微血管内皮细胞间紧密连接结构改变;应用Western blot技术分析小细胞肺癌细胞与微血管内皮细胞单层共培养时紧密连接蛋白occludin的可溶性片段和不可溶性片段在内皮细胞紧密连接处的分布情况;应用跨内皮迁移实验及辣根过氧化物酶渗漏实验分析小细胞肺癌细胞与内皮细胞单层共培养后其跨内皮细胞单层的迁移能力和内皮单层渗透性的改变;利用ROCK特异性抑制剂Y27632,分析Rho/ROCK信号通路是否参与小细胞肺癌细胞跨内皮单层迁移。结果小细胞肺癌细胞能够通过人脑微血管内皮细胞单层迁移,在0h到12h之间随培养时间的延长小细胞肺癌细胞的迁移率逐渐增加;共培养8h时人脑微血管内皮细胞紧密连接处紧密连接结构蛋白(ZO-1和occludin)表达减少,通透性明显增加;ROCK特异性抑制剂Y27632能够阻断小细胞肺癌细胞引起的内皮细胞间紧密连接处紧密连接结构蛋白ZO-1和occludin表达减少、内皮单层通透性的增加及小细胞肺癌细胞跨内皮单层迁移。结论小细胞肺癌细胞与人脑微血管内皮细胞单层共培养诱发人脑微血管内皮细胞间紧密连接的开放,利于小细胞肺癌细胞的跨内皮单层迁移。
Objective It is believed that brain metastases is easy to take place in patients with small cell lung cancer,however,the molecular mechanism is not well understood. The present study is to explore if the tight junction（TJ） of human brain mierovascular endothelial cells （HBMECs） was opened by small cell lung cancer ceils. Methods TJ proteins（ZO-1 and occludin） expression in HBMECs monolayer as measured by immunofluorescence assay, occludin＇s insoluble and soluble fractions expression was analyzed by Western blot.The transendothelial migration ability of small cell lung cancer cells and permeability of endothelial cells monolayers were analyzed by transendothelial migration assay and horseradish peroxidase（HRP） flux. The effect of Rho/ROCK signalling in the transendothelial migration of small cell lung cancer cells was investigated by Y27632 （ROCK inhibitor）. Rusults A time-dependent increase of transendothelial migration percentage of small cell lung cancer cells was observed in 0-hour to 12-hour period. The transendothelial migration increase of small cell lung cancer cells is associated with the increased permeability of HBMECs monolayer and the decreased expression of TJ proteins（ZO-1 and occludin）. Y27632 could decrease transendothelial migration ability of small cell lung cancer cells and permeability of HBMECs monolayer, and upregulate the expression of TJ proteins（ZO-1 and occludin） in HBMECs. Conclusion The small cell lung cancer cells promote TJ opening of HBMECs and the transendothelial migration of small cell lung cancer cells.
出处
《解剖科学进展》
CAS
2009年第2期183-187,191,共6页
Progress of Anatomical Sciences
基金
国家自然科学基金资助项目(No.30600270)
关键词
小细胞肺癌细胞
人脑微血管内皮细胞
共培养
紧密连接
small cell lung cancer cells
haman brain microvascular endothelial cells
cocuhivation
tight junction
