SODIUM BUTYRATE EXERTS AN ANTI-COLORECTAL CANCER ACTION BY INDUCING FERRITINOPHAGE IN HCT-116 CELLS
Objective To investigate whether sodium butyrate(NaB)can exert its anti-colorectal cancer action by inducing ferritinophagy.Methods The effects of NaB on the proliferation and death of colorectal cancer HCT-116 cells were investigated using MTT assay,plate cloning assay,and PI staining.The induction of ferritinophagy in HCT-116 cells by NaB was explored through the use of autophagy detection kit(MDC method),ferrous ion fluorescent probe FerroOrange.Western blot analysis was performed to detect the expression of p62,FTH1,LC3 and NCOA4 molecules.Results The results of the MTT and colony formation experiments showed that 2 mmol/L NaB significantly inhibited the viability(P<0.001)and clone formation(P<0.01)of HCT-116 cells.The PI staining results showed that NaB increased the number of dead cells,and the iron chelator deferoxamine(DFO)reversed the inhibitory and pro-apoptotic effects of NaB on HCT-116 cells(P<0.05).The autophagy inhibitor chloroquine(CQ)further enhanced the inhibitory effect of NaB on HCT-116 cells(P<0.001).The results of MDC method and FerroOrange detection showed that NaB increased the formation of autophagic vacuoles and intracellular Fe2+levels in HCT-116 cells(P<0.001).The upregulation of intracellular Fe2+levels by NaB was reversed by DFO and CQ(P<0.05).Western blot analysis revealed that NaB downregulated the protein expression levels of p62 and FTH1(P<0.001)and upregulated the protein expression levels of LC3-II,NCOA4 in HCT-116 cells(P<0.001,P<0.05).The downregulation of FTH1 protein expression level by NaB was reversed by CQ(P<0.05).Conclusion NaB can exert an anti-colorectal cancer action by inducing ferritinophagy in HCT-116 cells.
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