Abstract
Objective To investigate the roles ofc-Jun/Fral dimer in viability of dopaminergic neurons. Methods （1） Dopaminergic neuron-like cell line SH-SYSY was cultured in vitro. Cells were lysed for immunoprecipitation to determine whether c-Jun dimerized with Fral. （2） Cells received treatments were divided into four groups： Jun N-terminal kinase （JNK） inhibitor SP600125 treatment group, mitogen-activated kinase （MEK） 1/2 inhibitor U0126 treatment group, SP600125＋U0126 treatment group and control group （treated with dimethyl sulfoxide [DMSO]）. Western blotting was performed to detect the c-Jun and Fral expression levels and MTT assay was used to observe the cell viability. （3） Cells infected with adenovirus （Ad） carrying c-Jun or Fral genes were divided into three groups： Ad-c-Jun group, Ad-Fral group and Ad-c-Jun＋Ad-Fral group, and cells transfected with GFP were used as control group; immunofluorescence and MTT assay were performed to determine the infectious rate and the cell viability, respectively. Results （1） Co-immunoprecipitation showed that the c-Jun dimerized with Fral in SH-SYSY cell line. （2） Western blotting indicated that inhibition of JNKby SP600125 reduced c-Jun expression and MEK1/2 by U0126 inhibited Fral expression; MTT assay indicated that there were significant differences in the cellular viability among the four groups （F=I 6.647, P=-0.000）, the cellular viability in control group obviously differed from that in SP600125 treatment group, U0126 treatment group or SP600125＋U0126 treatment group （P〈0.05）. （3） Immunofluorescence showed that most of cells infected by adenovirus expressed c-Jun or Fral. MTT assay indicated that there were significant differences in the cellular viability among the four groups （F=14.543, P=-0.000）, and there were significant differences in their viabilities between control group and Ad-Fral group, and between Ad-Fral group and Ad-Fral＋Ad-c-Jun group （P〈0.05）. Conclusion The c-Jun and Fral forming a dimer promotes the viability of dopaminergic neuron-like cells. Key words: Dopaminergic neuron-like cell; C-Jun; Fral; Viability
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