Exploring the role of ferroptosis in the resistance of liver cancer cells to lenvatinib based on RNA-seq technology
Objective To explore the role of ferroptosis in the resistance of liver cancer cells to lenvatinib(Lenva),and provide new ideas for preventing or reversing Lenva resistance.Methods Human liver cancer cell line PLC/PRF/5 cells also known as parental cells(PC)were cultured in vitro,and Lenva resistant cells(LRG)were constructed using gradient concentration induction method.RNA-seq technology was used for transcriptome sequencing of LRC and PC.Differential expression gene analysis and KEGG enrichment analysis were completed.Cell viability was measured by CCK-8 assay.Expressions of ferroptosis related proteins were detected by Western blot.The reactive oxygen species(ROS)was detected using DHE probes,and changes in mitochondrial morphology were observed by transmission electron microscopy(TEM).Results The half inhibitory concentration IC50 of Lenva in PC group and LRC group was 24.88 μmol/L and 89.34 μmol/L,respectively,indicating the successful induction of Lenva resistant human liver cancer cells in vitro.A total of 12106 genes were detected by RNA-seq,among which 88 genes were upregulated and 197 genes were downregulated in the LRC group compared to the PC group.The KEGG pathway enrichment analysis of differential expression genes showed that ferroptosis signaling pathway was enriched within it.Under the exposure of Lenva(10 μmol/L for 48 h),the expression of ferroptosis related proteins SLC7A11 and GPX4 were increased in the LRC group compared to the PC group,while the content of ROS was decreased.TEM results showed that compared to the PC group,the characteristic mitochondrial morphological changes related to ferroptosis in the LRC group were significantly reduced.Conclusion Lenva resistance in liver cancer cells may involve multiple genes and signaling pathways,among which ferroptosis resistance may be a key role in causing Lenva resistance.
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