The Effect Of Rapamycin On Proliferation Of Human Keloids Fibroblasts In Vitro

Objective To investigate the effect of rapamycin on proliferation of Human keloid fibroblasts in vitro, to further explore the mechanism of it.Methods With different concentrations of rapamycin effect on cultured keloid fibroblasts, and CCK-8 assay was adopted to evaluate cell survival. Immunohistochemistry(10 nmol/L), RT-PCR and western blot(0.1n mol/L,1 nmol/L,10 nmol/L,100 nmol/L) were performed to detect the expression of P70s6k and 4E-BP1 before and after intervention in cultured keloid fibroblasts, cultured normal fibroblasts.Result CCK-8 assay detected the absorbance of keloid fibroblasts, which decreased with the concerntrations and time increased.There were significant differences between these groups (P<0.05).IHC(rapamycin concentration 10nmol/L intervention), RT-PCR and western blot results showed that gene expression and protein expression of P70s6k and 4E-BP1 were all decreased in(0.1nmol/L,1nmol/L,10nmol/L,100nmol/L)rapamycin concentration before and after intervention.The level of keloid's protein and mRNA expression between P70s6k and 4E-BP1 was much higher than the normal's.There were significant differences between these groups (P<0.05).Conclusions rapamycin significantly inhibited the proliferation of Human Keloids Fibroblasts cell in vitro. rapamycin also inhibited the expression of P70s6k and 4E-BP1.The expression reduce of P70s6k and 4E-BP1 as well as the dependablity between P70s6k and 4E-BP1 maybe were the mechanism of abnormal keloid's formation.