uPAR mRNA测定及其在U937细胞中的表达

Determination of u - PAR mRNA and Its Expression in U937 Cells

目的研究凝血酶、TNF－α.IFN-γ等对U937细胞表达uPARmRNA的影响，观察凝血系统、炎症介质与纤溶功能变化间的关系。方法建立并应用逆转录PCR（RT－PCR）法测定培养U937细胞的uPARmRNA。结果1．经测定批间变异系数（CV间）确认了RT－PCR法测定uPARmRNA的稳定性；2以不同浓度凝血酶刺激24h，对U937细胞表达uPARmRNA有抑制和促进两种不同的效应：TNF－α、IFN－γ、ATRA和As2O3都能促进uPARmRNA表达，但剂量-效应关系并不相同。结论与凝血和炎症反应相关的因子及某些药物以不同方式和程度影响细胞表达uPARmRNA，说明它们的作用机制和意义并不相同。

Obiective This study was designed to observe the expressive changes ofurokinase - type plasminogen activator receptor (u - PAR) mRNA in cultured U937 cellsunder the conditions of adding thrombin, TNF - a, IFN - γ, all - trans - retinoic acid (ATRA)or As2O3 into the experimental cell cultures. Metbods The method of reverse transcriptio-nal PCR (RT- PCR) for examination of u- PAR mRNA in cultured U937 cells has beenestablished and has been used to monitor the u - PAR mRNA levels in all the experiments.Rcsults The coefficients of variation for between - assay were obtained from O.97 % to 8.59%with the method of RT- PCR for determination of u - PAR mRNA through four groups ofexperiments. The u - PAR mRNA levels showed up - expressed by actions of the agents ofTNF - a, IFN -γ, ATRA and As2O3 respectively in different doses on the cultured cells withincertain concentration ranges. Changes of expression were different between differentagent - experimental groups. When U937 cells culturedin in medium with thrombin upto 24h,the expression of u - PAR mRNA increased dose - dependently in the thrombin concentra-tions from O.05 to 1.0nmol/L, and it showed significantly decreased with higherconcentrations of thrombin up to 40nmol/L. Conclusion The phenomenon showed by theresults may be related to that there were some different mechanisms responsible for theseagents which contribute to the expression of u - PAR mRNA by U937 cells.