Figure 4.

FASN regulates proliferation, migration, apoptosis, and lipid droplets formation of human ccRCC cells. (A,B) After transient transfected with FASN plasmid in normal control, FASN-RNAi1, FASN-RNAi2, and FASN overexpression (OE) groups, we investigated FASN mRNA and protein expression level in A498 and 786O cells and found significant differential expression levels in the RNAi1, RNAi2, and OE groups compared with A498 and 786O cells. (C,D) After the A498 and 786O cells were transfected for 5 days, the A450 OD values revealed that cells proliferation ability was significantly suppressed in the RNAi1 and RNAi2 groups and significantly promoted in the OE groups compared with normal control (p < 0.05). (E,F) The X and Y axes represent Annexin V-FITC and PI, respectively. Apoptotic cells number was increased in the RNAi1- and RNAi2-transfected groups compared with control in 786O cells, whereas the FASN OE groups showed similar apoptosis to normal control cells. (G,H) To explore the role of FASN-mediated migration ability of ccRCC cells, we performed transwell migration assay and found that the inhibition of FASN markedly restrained migrated cell numbers. Meanwhile, the OE of FASN significantly promotes migrated A498 and 786O cell counts. (I) To explore whether FASN regulates ccRCC cells lipid formations, we performed Oil Red O staining and found that inhibition of FASN significantly reduced absorption of lipid droplets in lipid-rich cells. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.