The Effects Of Ulinastatin On The UPA Expression And Invasion Of Choiocarcinoma

Objective To detect the expression of urokinase-type plasminogen activator (uPA) in different invasion ability human choriocarcinoma cell lines JEG-3 and JAR,and observe the effects of Ulinastatin on the level of uPA gene expression and cell invasion in vitro of JEG-3 cells, to investigate the correlation between the expression of uPA mRNA and the invasion and metastasis ability of choriocarcinoma,and explore the anti-invasion and metastasis and its mechanism of Ulinastatin in choriocarcinoma.Methods (1) Culture choriocarcinoma cell lines JEG-3 and JAR in vitro; (2) To detect the invasive ability of JEG-3 and JAR cells by Matrigel invasion assay, using semi-quantitative RT-PCR (reverse transcriptase-polymerase chain reaction) to detect the expression of uPA mRNA in JEG-3 and JAR; (3) The high-invasive cell line JEG-3 were cocultured respectively with OU/ml, 50U/ml,100U/ml,200U/ml,400U/ml,800U/ml Ulinastatin for 24 hours, the effects of Ulinastatin on the expression of uPAmRNA and the invasion of JEG-3 cells were measured by RT-PCR and Matrigel invasion assay.Results (1) The invasion ability of JEG-3 cell line was stronger than that of JAR cell line(P<0.05); the expression of uPA gene in JEG-3 cells were significantly stronger than that in JAR cells (P< 0.05);(2) The expression of uPA mRNA in 50U/ml group has no significant difference compared with the control group(OU/ml) (P>0.05); when the Ulinastatin concentration at 100U/ml and above, the differences were statistically significant compared with the control group(P<0.05), and the differences between any two groups were significantly (P<0.05); With the increasing concentration of Ulinastatin,the level of uPAmRNA expression was decreasing, correlation analysis showed there is a negative correlation between uPA mRNA levels and Ulinastatin concentration (r=-0.862, P<0.05);(3) Trans-membrane number of JEG-3 cells was reduced in all groups; there is no significant difference at 50U/ml group compared with the control group, (P>0.05); at 100U/ml and above, the differences were statistically significant compared with control group (P<0.05), and differences between any two groups were also statistically significant (P<0.05); the invasive ability of JEG-3 was weakening when the concentration of Ulinastatin was increasing, correlation analysis showed that the invasion ability and the Ulinastatin concentration was negatively correlated (r=-0.885,P<0.05), and the choriocarcinoma cell invasive ability and the level of uPA mRNA expression was positively correlated (r=0.996,P<0.05). Conclusion (1) The expression of uPA mRNA was obvious different between the high and low invasiveness choriocarcinono-ma cells,the level of uPAmRNA expression affect the invasiveness of choriocarcinoma cells; (2) Ulinastatin can down-regulate the expression of uPA in the high-invasive JEG-3 cells; (3) Inhibiting the expression of uPAmRNA can reduce the invasiveness of JEG-3 cells in vitro.