Fig. 7
TRIM6 regulates Snail1 expression and HCC cell invasion via DDX58. (A) Immunoblot analysis of Snail1 protein levels in HCCLM3 cells following TRIM6 knockdown, with or without concomitant DDX58 knockdown. Silencing TRIM6 led to a reduction in Snail1 protein levels, which was partially restored upon co-knockdown of DDX58. β-Actin served as a loading control. (B) Quantitative RT-PCR analysis of Snail1 mRNA levels under the same conditions as in (A). TRIM6 knockdown reduced Snail1 transcript levels, which were partially rescued by simultaneous DDX58 knockdown. **p < 0.01, ***p < 0.001, ns = not significant. (C, D) Transwell invasion assay evaluating the invasive capacity of HCCLM3 cells following TRIM6 knockdown, with or without DDX58 knockdown. (C) Representative images of invaded cells stained with crystal violet. (D) Quantification of invasive cell numbers. TRIM6 silencing significantly impaired cell invasion, which was partially restored by DDX58 knockdown. **p < 0.01, ***p < 0.001. (E) Immunoblot analysis of Snail1 protein levels in HCCLM3 cells overexpressing TRIM6, with or without DDX58 overexpression. TRIM6 overexpression markedly elevated Snail1 protein expression, which was attenuated by co-expression of DDX58. β-Actin was used as a loading control. (F) Quantitative RT-PCR analysis of Snail1 mRNA levels in HCCLM3 cells upon TRIM6 overexpression, with or without DDX58 overexpression. The TRIM6-induced upregulation of Snail1 transcripts was reversed by enforced expression of DDX58. **p < 0.01, ***p < 0.001, ns = not significant. (G, H) Transwell invasion assay assessing the effects of TRIM6 and DDX58 overexpression on HCCLM3 cell invasiveness. (G) Representative images of invading cells stained with crystal violet. (H) Quantification of invasive cell numbers. TRIM6 overexpression significantly enhanced invasion, which was substantially suppressed by DDX58 overexpression. ***p < 0.001, ****p < 0.0001.
