凝血酶诱导非小细胞肺癌血管内皮生长因子分泌的实验

Inducible Effect of Thrombi on Secretion of Vascular Endothelial Growth Factor in Non-small Cell Lung Carcinoma

目的探讨凝血酶对非小细胞肺癌细胞株A549分泌血管内皮生长因子(Vascular endothelial growth factor,VEGF)的促进作用及其机制。方法采用ELISA法检测凝血酶对A549细胞分泌VEGF的浓度以及水蛭素对其阻断作用。采用RT-PCR法检测凝血酶受体PAR-1在A549细胞中的表达。结果凝血酶在0.5～3U/ml的浓度范围内可以明显地增加VEGF的分泌,而<0.5U/ml时这种促进作用则不明显,水蛭素能够完全阻断凝血酶的这种促进作用。PAR-1在A549细胞中表达,而在正常的胎肺细胞(KMB-17)中并不表达。结论在一定浓度范围内,凝血酶能有效地促进A549细胞VEGF的分泌,而这种促进作用可能与PAR-1受体的表达有关。

Objective To study thrombin for the up-regulation of vascular endothelial growth factor on the human non-small cell lung cancer cells line A549, and to explore its mechanism. Methods The ELISA methods was used to evaluate the role of thrombin on the expression of vascular endothelial growth factor （VEGF） in A549 and inhibition of hirudin. RT-PCR was used to evaluate the expression of thrombin receptor （PAR-1） in A549 cell. Results Thrombin may increase the secretion of VEGF in A549 cell in 0. 5 -3 U/ml concentrations. The induction of VEGF was significantly blocked by the thrombin inhibitor hirudin. RT-PCT demonstrated PAR-1 expressed in A549 cell but not in KMB-17. Conclusion Thrombin may substantially stimulate the secretion of VEGF in A549 cell within a certain degree. The mechanisms of induction were might related with the expression of thrombin receptor PAR-1.