Inhibition of uPA Expression with siRNA in Mice Sertoli Cells

Abstract: The aim of our study is to analyze the suppressive effects of siRNA targeting uPA gene (siuPA) in uPA expression in mice sertoli cell line (TM4 cells). 3 different sequences of si-uPA were transfected into TM4 cells to screen the effective si-uPA. The inhibition of uPA expression was observed at various time points after transfection with the effective si-uPA in three different concentration (30， 50， 100 nmol/L). The results showed that the proper concentration of transfection for TM4 cells was 50 nmol/L. Among the three si-uPA， si-uPA1 had best suppressive effect. After transfection of si-uPA1 into TM4 cells， the decrease of uPA mRNA expression was observed in all three groups at 24 h， and the suppressive rate reached 70% in 100 nmol/L group， which was much obvious than those in 30 nmol/L and 50 nmol/L groups (P<0.05). The relative expression of uPA reduced steadily with the extension of transfection. At 72 h after transfection， the uPA mRNA in those three groups were 53.9%， 35.3% and 27.7%， respectively， compared with those in blank control (P<0.05). In this study， the effective sequence of siRNA targeting uPA gene was screened from three candidates and the inhibition effect continued to 72 h. The suppressive effects of the same siRNA concentration showed no significant differences when detected at different time points， while variant concentration of siRNA inhibited the expression of uPA gene in a dose-dependent manner.

CSTR: 32200.14.cjcb.2012.10.0007