Effects Of Calpastatin Peptide,Calpain Inhibitor ? And Calpeptin On Expression Of AP-1 And PARP Of C2C12 Myoblasts In Tumor Microenvironment

Objective To explore the effects of 3 kinds of calpain inhibitor on expression of AP-1 and PARP of C2C12 myoblasts in tumor microenvironment.Methods C2C12 myoblasts and CT26 cells were co-cultured by transwell in 6-well plate,then divided into 5 groups: CalP(Calpastatin Peptide),Cal4(calpain inhibitor IV),Cal(Calpeptin),DmD(DMSO)and Dm S(DMSO,C2C12 myoblasts only).After intervention of certain time,immunocytochemistry was used to detect protein levels of AP-1 and PARP,and quantitative real-time polymerase chain reaction was used to detect mRNA levels.Results immunocytochemistry:CalP reduces the total protein levels of AP-1 at 6h to 24 h.Cal4 reduces the total protein level of AP-1 at 24 h.There is no obvious change of total protein level of PARP after intervention of CalPCal4,and Cal.QPCR: Cal P,Cal4 down-regulates the mRNA levels of AP-1 at 6h,12 h,48h,72 h.Cal down-regulates the mRNA levels of AP-1 from 6h to 72 h.CalP up-regulates the mRNA level of PARP at 6h and down-regulates it at 12 h and 72 h.Cal4down-regulates the mRNA level of PARP at 12 h,24h and 72 h.Cal up-regulates the mRNA level of PARP at 48 h and down-regulates it at 12 h and 72 h.Conclusion Calpain inhibitors can down-regulate the expression of AP-1,and manifestation of biological effects after drugs intervention is anti-cancer.Intervention of Calpain inhibitors makes no difference of expression of PARP.