Effects of NEAT1 on expression of GPX4 through IGF2BP2 on middle ear cholestatoma cells
Objective To investigate the effect of nuclear-enriched abundant transcript 1(NEAT1)on human middle ear cholesteatoma cells through the regulation of glutathione peroxidase 4(GPX4)expression by insulin-like growth factor 2 mR-NA-binding protein 2(IGF2BP2).Methods Intraoperative specimens and paraneoplastic normal tissue specimens of 30 pa-tients with middle ear cholesteatoma collected from the Department of Otorhinolaryngology of the First Affiliated Hospital of Zhengzhou University during the period from May 2021 to December 2022 were selected and were ordered as the control group and middle ear cholesteatoma group,respectively.Cultured ear cholesteatoma cells were transfected with NEAT1 overexpres-sion plasmid,small interfering RNA,disordered siRNA and PBS,respectively.They were named as NEAT1 group,si-NEAT1 group,si-RNA-negative control(si-NC)group and normal control(NC)group,respectively.The expression levels of NEAT1,IGF2BP2,and GPX4 were determined by real-time quantitative polymerase chain reaction(qRT-PCR).Dual luciferase reporter gene assay verified the targeting relationship between NEAT1 and IGF2BP2.The effect of NEAT1 on chol-esteatoma cell activity was verified using CCK-8 assay.Results The relative expression of NEAT1 in the cholesteatoma group was higher than that in the control group,the difference was statistically significant(P＜0.05).Compared with the NC group,the relative expression of NEAT1 was elevated in the NEAT1 group and decreased in the si-NEAT1 group,the differ-ence was statistically significant(P＜0.05).The results of dual luciferase reporter gene assay showed that the relative lucifer-ase activity of the cholesteatoma cell in the IGF2BP2 mimics+NEAT1-WT group was lower than that in the miRNA-NC+G-CSF-WT group,the difference was statistically significant(P＜0.05).There was no statistically significant difference in the relative luciferase activity between the IGF2BP2 mimics+NEAT1-MUT group compared with the miRNA-NC+NEAT1-MUT group(P＞0.05).Compared with the NC group,the relative expression of IGF2BP2 and GPX4 mRNA was decreased in the NEAT1 group,and the relative expression of IGF2BP2 and GPX4 mRNA was elevated in the si-NEAT1 group,the difference was statistically significant(P＜0.05).Compared with the NC group,cell activity increased in the NEAT1 group and decreased in the si-NEAT1 group,the difference was statistically significant(P＜0.05).Conclusion NEAT1 enhances middle ear cholesteatoma cell activity by inhibiting the IGF2BP2/GPX4 pathway,and NEAT1 can be investigated as a thera-peutic target for ear cholesteatoma.
Cholesteatoma of middle earLong non-coding RNANuclear-enriched abundant transcript 1Insulin-like growth factor 2 mRNA-binding protein 2Glutathione Peroxidase 4
NETL
NSTL
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