Fig. 6

FTO inhibits HCC by downregulating VEGFA expression. (a) Transcriptional sequencing of FTO-overexpressing MHCCLM3 cells compared to the control group revealed significant downregulation of 309 genes and upregulation of 227 genes. Differentially expressed genes were computed using the negative binomial generalized linear model in edgeR. (b) GO analysis of the differentially expressed genes unveiled ten major functional pathways and 78 key genes. (c) Further analysis indicated that the cellular biological processes primarily affected by FTO overexpression were lipid metabolism, angiogenesis, and hypoxia. (d) Results from protein chip detection showed an inverse trend between VEGFA expression and FTO alteration. Knockdown of FTO in MHCC97L cells led to upregulation of VEGFA, while overexpression of FTO in MHCC97H cells resulted in significant downregulation of VEGFA. Differences were assessed using an unpaired Student’s t-test. (e) Western blot analysis indicated a significant upregulation of VEGFA expression in FTO-knockdown HepG2 cells, whereas FTO-overexpressing MHCCLM3 cells exhibited a pronounced downregulation of VEGFA expression. (f) We reversed of the FTO-induced downregulation of VEGFA expression through VEGFA plasmid. The results of the scratch assay and transwell assay demonstrated that the invasive and migratory abilities of HCCLM3 were significantly enhanced after the restoration of VEGFA expression. (g) VEGFA expression was reduced in FTO-upregulated 97 L cells through VEGFA siRNA, and the results of the scratch assay and transwell assay revealed a corresponding decrease in the invasive and migratory abilities of 97 L cells. Differences were assessed using an unpaired Student’s t-test. *P<0.05