Androgen-induced gene 1 correlates with extracellular vesicle-associated cholesterol metabolism and hepatic metastasis of pancreatic cancer

Figure 6 Androgen-induced gene 1 knockdown rewires cholesterol metabolism in tumor cells. A: Volcano plot of differentially expressed genes identified by bulk RNA sequencing comparing androgen-induced gene 1 knockdown (AIG1-KD) and non-targeting control single-guide RNA (sgControl) cells. Genes not meeting both criteria are labeled as “none”. Selected upregulated and downregulated genes relevant to inflammatory signaling, cholesterol metabolism, tumor-suppression and tumor-promotion are highlighted; B: Kyoto Encyclopedia of Genes and Genomes enrichment analysis of downregulated genes. The Q-value shown in the plot represents the false discovery rate-adjusted P-value. Cholesterol metabolism was the only significantly enriched pathway among downregulated genes (false discovery rate-adjusted P < 0.05); C: Heatmap showing the relative expression of cholesterol metabolism-related genes between groups based on bulk RNA-sequencing analysis; D: Quantitative real-time polymerase chain reaction validation of cholesterol metabolism-related genes in sgControl and AIG1-KD cells; E: Total cholesterol, free cholesterol, and free cholesterol/total cholesterol ratio measured in sgControl and AIG1-KD cells; F: Comparison of cholesterol efflux between the two groups. Data are presented as mean ± SD of three independent experiments. Statistical analysis was performed using unpaired t-tests. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001. KEGG: Kyoto Encyclopedia of Genes and Genomes; CAM: Cell adhesion molecule; sgControl: Non-targeting control single-guide RNA; AIG1-KD: Androgen-induced gene 1 knockdown; HMGCR: 3-hydroxy-3-methylglutaryl-coenzyme A reductase; DHCR24: 24-dehydrocholesterol reductase; LRP2: Low-density lipoprotein receptor-related protein 2; ABCA1: ATP-binding cassette subfamily A member 1; APOE: Apolipoprotein E; SREBF2: Sterol regulatory element binding transcription factor 2; FC: Free cholesterol; TC: Total cholesterol.