IGF2BP1 promotes gastric cancer progression by stabilizing HS6ST2 mRNA in an N6-methyladenosine-dependent manner

Figure 3 Insulin-like growth factor 2 mRNA-binding protein 1 promotes tumor growth via an N6-methyladenosine-dependent mechanism. A and B: Subcutaneous xenograft tumor model was used to evaluate the effects of insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) on tumor growth in vivo. Representative tumor images (A); tumor growth curves and final tumor weight statistics (B); C: Venn diagram of multi-omics analysis identifies heparan sulfate 6-O-sulfotransferase 2 (HS6ST2) and SH3-domain binding protein 1 as core IGF2BP1 target genes linked to poor prognosis; D: Positive correlation between IGF2BP1 and HS6ST2 mRNA expression in The Cancer Genome Atlas-stomach adenocarcinoma; E: High HS6ST2 expression correlates with poor overall survival in The Cancer Genome Atlas-stomach adenocarcinoma; F: The expression of the HS6ST2 protein in gastric cancer tissues compared with adjacent non-tumor tissues; G: Predicted N6-methyladenosine (m6A) modification sites on HS6ST2 mRNA (sequence-based RNA adenosine methylation site predictor tool); H: M6A enrichment of HS6ST2 mRNA in SGC7901 and MGC803 cells (MeRIP-quantitative polymerase chain reaction); I and J: M6A levels of HS6ST2 mRNA are decreased upon IGF2BP1 knockdown (I) and increased upon its overexpression (J); K and L: IGF2BP1 overexpression increases the luciferase reporter activity driven by the wild-type HS6ST2 3’UTR (K), an effect that is lost upon mutation of the predicted m6A site (L). Data are expressed as mean ± SD. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. IGF2BP: Insulin-like growth factor 2 mRNA-binding protein; shRNA: Short hairpin RNA; NC: Negative control; TPM: Transcripts per million; HR: Hazard ratio; HS6ST2: Heparan sulfate 6-O-sulfotransferase 2; m6A: N6-methyladenosine; RIP: RNA immunoprecipitation.