姜黄素通过调控PI3K/AKT通路诱导骨肉瘤细胞焦亡OA

AIM:To investigate the effect of curcumin on pyroptosis in human osteosarcoma U2OS cells and MG63 cells.METHODS:The U2OS and MG63 cells were cultured in vitro and divided into a control group and groups treated with different concentrations(20,40,60 and 80 µmol/L)of curcumin.Cell viability was assessed using the MTT assay,lactate dehydrogenase(LDH)levels were measured by colorimetric assay,and cellular morphology was examined by scanning electron microscopy.Levels of interleukin-1β(IL-1β)and IL-18 in the cell culture supernatants were quanti-fied by ELISA.Western blot analysis was performed to determine the expression levels of cleaved caspase-1,N-terminal fragment of gasdermin D(GSDMD-N),nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),as well as key proteins involved in the PI3K/AKT signaling pathway.RESULTS:Compared with the control group,curcum-in treatment at various concentrations significantly inhibited the viability of both U2OS and MG63 osteosarcoma cells(P<0.05),with IC50 values of 36.4 and 31.7 µmol/L,respectively.Treatment with 20 and 40 µmol/L curcumin markedly in-creased LDH release into the cell culture supernatant(P<0.05),indicating enhanced membrane permeability.Scanning electron microscopy revealed characteristic features of pyroptosis in these groups,including cellular swelling and plasma membrane rupture.Furthermore,compared with control group,both 20 and 40 µmol/L curcumin groups exhibited signifi-cantly elevated levels of IL-1β and IL-18 in the supernatant(P<0.05).Consistently,Western blot analysis showed that curcumin(20 and 40 µmol/L)significantly up-regulated the expression of key pyroptosis-related proteins,cleaved cas-pase-1,GSDMD-N and NLRP3,in both U2OS and MG63 cells.In addition,curcumin treatment(20 and 40 µmol/L)sig-nificantly suppressed the PI3K/AKT signaling pathway,as evidenced by a marked decrease in the phosphorylated AKT(p-AKT)/total AKT ratio and reduced PI3K protein expression compared with the control group.CONCLUSION:Curcumin induces pyroptosis and suppresses the survival of osteosarcoma U2OS and MG63 cells,at least in part,through inhibition of the PI3K/AKT signaling pathway.