Fig. 5

5-lipoxygenase drives ccRCC migration phenotype and promotes sensitivity to ferroptosis. A ALOX family gene expression in ccRCC tumor biopsies (denoted as T) compared to non-malignant renal cortex (denoted as N). Data curated from TCGA KIRC dataset. Comparisons are multiple comparisons from a one-way ANOVA using Šídák’s multiple comparisons test to measure the differences between all groups. 11 degrees of freedom. P(ALOX5 N vs ALOX5 T) = < 0.0001. P(ALOX5AP N vs ALOX5AP T) = 0.002. B Kaplan-Meyer plots of overall survival in patients expressing high (upper quartile) or low expression of 5-LOX pathway genes. Dashed lines represent 95% confidence intervals. C qPCR measurement of relative mRNA expression of ALOX5 in ccRCC cells transfected with siRNA against either ALOX5 or a control scrambled siRNA (siCTRL). Bar graph represents mean and error bars represent standard deviation. n = 2 with 3 technical replicates per experiment. Comparisons are from an unpaired two-tailed Student’s t test. ***p < 0.001. D, E Cell titer blue viability assay of ccRCC cells (M62 or RCC4 cells as indicated) cultured with 5-LOX inhibitor zileuton or transfected with siALOX5 and increasing concentrations of erastin. n = 3 biological replicates, error bars depict the standard deviation from the mean. F Summary model of the role of ACSL3 and 5-LOX in ccRCC as well as compounds effecting the system. Loss of functional pVHL leads to a change in metabolism that leads to an increase in lipid uptake as well as a decrease in beta oxidation of lipids, resulting in lipid droplet accumulation in the cytoplasm. ACSL3 is required to process lipids which eventually leads to downstream PUFA metabolism that leads to the production of lipid hydroperoxides. These lipid peroxides can sensitize the cell to ferroptosis if the cell is unable to properly detoxify them