Research Paper Volume 13, Issue 9 pp 13300—13317

SnoRD126 promotes the proliferation of hepatocellular carcinoma cells through transcriptional regulation of FGFR2 activation in combination with hnRNPK

SnoRD126 combining with hnRNPK to up-regulate FGFR2. (A) qRT-PCR assay for FGFR2 expression in HepG2 cell and snoRD126-overexpressing Huh7 cells after transfected with hnRNPK siRNAs. (B) qRT-PCR assay for FGFR2 expression in Huh7 cells with snoRD126 WT or mutants-overexpressing. (C) Overexpression of snoRD126 rather than the mutants activated phosphorylation of AKT and p70S6K in Huh7 cells as measured by immunoblotting. (D) Immunoblotting was performed for the indicated proteins in snoRD126- or vector-overexpressed Huh7 cells with hnRNPK siRNA treatment. (E, F) ChIP assay using hnRNPK antibody followed by qRT-PCR assay for hnRNPK occupancy at FGFR2 promoter region in (E) HepG2 and (F) snoRD126-overexpressed Huh7 cells. Four primer pairs were used to assess the occupancy at every 250 bp upstream of FGFR2 transcription start site (TSS). (G) Luciferase reporter assays for FGFR2 promoter activity after transiently transfected hnRNPK in HEK-293 cells, mean ± SD, **PC, D), loading control.

Figure 4. SnoRD126 combining with hnRNPK to up-regulate FGFR2. (A) qRT-PCR assay for FGFR2 expression in HepG2 cell and snoRD126-overexpressing Huh7 cells after transfected with hnRNPK siRNAs. (B) qRT-PCR assay for FGFR2 expression in Huh7 cells with snoRD126 WT or mutants-overexpressing. (C) Overexpression of snoRD126 rather than the mutants activated phosphorylation of AKT and p70S6K in Huh7 cells as measured by immunoblotting. (D) Immunoblotting was performed for the indicated proteins in snoRD126- or vector-overexpressed Huh7 cells with hnRNPK siRNA treatment. (E, F) ChIP assay using hnRNPK antibody followed by qRT-PCR assay for hnRNPK occupancy at FGFR2 promoter region in (E) HepG2 and (F) snoRD126-overexpressed Huh7 cells. Four primer pairs were used to assess the occupancy at every 250 bp upstream of FGFR2 transcription start site (TSS). (G) Luciferase reporter assays for FGFR2 promoter activity after transiently transfected hnRNPK in HEK-293 cells, mean ± SD, **P<0.01, NS, not significant. GAPDH and β-actin in (C, D), loading control.