The expression of Selenoprotein W(SelW)in C2C12 skeletal muscle cells was specifically decreased to examine its influence on the amount of glutathione(GSH)and the activity of glutathione peroxidase(GPx).SelW knock-down was performed by RNA interference(RNAi)in cultured muscle cells and verified by Real-time PCR and Western blotting.In addition,cell viability,GSH content and GPx activity were assayed.The results showed that the mRNA level and protein expression of SelW were decreased successfully by 71.9%and 68.8%relative to control values,cell viability decreased by 21.5%,GSH increased by 29.76%,and GPx increased by 47.58%.WST assay showed that compared with blank control,the value of positive group dropped 21.5%;In GSH and GPx assay,compared with blank control the positive group increased29.76%and 47.58%separately.In conclusion,SelW knock-down by RNAi caused significant cytotoxity in skeletal muscle cells and led to compensatory increases in GSH content and GPx activity.These findings are consistent with the suggestions from bioinformatics indicating an antioxidative role for SelW in skeletal muscle cells.
WANG Xiao-longXU KaiQIN Ou-juCHEN Xiao-fengYANG Chuan-ping