The mechanism of miR-32-5p mediated ferroptosis on thyroid cancer cell proliferation through the p53/SLC7A11 signaling pathway
Objective To investigate whether miR-32-5p can affect thyroid cell proliferation by regulating the p53/SLC7A11 signaling pathway mediated ferroptosis.Methods TPC-1 cells were randomly divided into control group,miR-NC group,miR-32-5p group,miR-32-5p+Fer-1 group,miR-32-5p+si-p53 group,and miR-32-5p+Fer-1+si-p53 group.RT-PCR was used to detect the relative expression level of miR-32-5p in cells;CCK-8 method for detecting cell proliferation;Detect the content of Fe2+,MDA,SOD,GSH,and ROS in cells;JC-1 method for de-tecting mitochondrial membrane potential levels in cells;Protein blotting was used to detect the expression of p53,GPX4,and SLC7A11 proteins in cells.Results The relative expression levels of miR-32-5p in thyroid cancer cell lines BCPAP,TPC-1,and SW1736 were lower than those in human normal thyroid cell line HT ori3 cells(P＜0.01).Compared with the Control group,the miR-32-5p group showed a significant increase in the relative expres-sion level of miR-32-5p,MDA content,Fe2+content,ROS fluorescence intensity,and p53 protein expression in cells.However,the cell survival rate,GSH content and SOD activity,mitochondrial membrane potential level,and GPX4 and SLC7A11 protein expression in cells were significantly reduced(P＜0.01);Compared with the miR-32-5p group,the miR-32-5p+Fer-1 group,miR-32-5p+si-p53 group,and miR-32-5p+Fer-1+si-p53 group showed a signifi-cant decrease in the relative expression of miR-32-5p,MDA content,Fe2+content,ROS fluorescence intensity,and p53 protein expression in cells.The cell survival rate,GSH content and SOD activity,mitochondrial membrane po-tential level,and GPX4 and SLC7A11 protein expression in cells were significantly increased(P＜0.01);The rela-tive expression level of miR-32-5p,MDA content,Fe2+content,ROS fluorescence intensity,and p53 protein expres-sion in the miR-32-5p+Fer-1+si-p53 group were significantly lower than those in the miR-32-5p+Fer-1 group.The cell survival rate,GSH content and SOD activity,mitochondrial membrane potential level,and GPX4 and SLC7A11 protein expression in the cells were higher than those in the miR-32-5p+Fer-1 group(P＜0.05).Conclusion Over-expression of miR-32-5p can inhibit thyroid cancer cell proliferation by promoting ferroptosis,which may be related to the regulation of the p53//SLC7A11 signaling pathway.
Thyroid cancermicroRNA-32-5p(miR-32-5p)Ferroptosis P53//SLC7A11 signaling path-wayProliferationTPC-1 cells line
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