Research Paper Volume 13, Issue 15 pp 19317—19338

The deubiquitinase OTUD3 stabilizes ACTN4 to drive growth and metastasis of hepatocellular carcinoma

OTUD3 downregulation inhibits HCC cells migration and invasion in vitro and in vivo. (A, B) Invasion and migration assays were employed to evaluate the effect of OTUD3 knockdown on HCCLM3 cells metastatic ability (Magnification 200X). (C, D) Scratching assay was performed to detect migration ability of OTUD3 knockdown HCCLM3 cells compared with the control group. (E) In vivo tumour metastasis was examined using the nude mice (n=6 per group) injected with luciferase-expressing HCCLM3 cells stably transfected with shNC or shRNA targeting OTUD3 and was detected by IVIS from day 0 to day 35. Representative images obtained are shown. (F) Quantification of metastatic lung nodules with shNC and shOTUD3 HCCLM3 cells by tail-vein injection. (G) Images of H&E staining of paraffin-embedded lung tissues from shNC and shOTUD3 nude mice group (Magnification: 100X, 200X, 400X). The unpaired two-sided Student’s t test was used for comparing between two groups of equal variances. Error bars represent mean ± SD from three independent experiments. *PP

Figure 3. OTUD3 downregulation inhibits HCC cells migration and invasion in vitro and in vivo. (A, B) Invasion and migration assays were employed to evaluate the effect of OTUD3 knockdown on HCCLM3 cells metastatic ability (Magnification 200X). (C, D) Scratching assay was performed to detect migration ability of OTUD3 knockdown HCCLM3 cells compared with the control group. (E) In vivo tumour metastasis was examined using the nude mice (n=6 per group) injected with luciferase-expressing HCCLM3 cells stably transfected with shNC or shRNA targeting OTUD3 and was detected by IVIS from day 0 to day 35. Representative images obtained are shown. (F) Quantification of metastatic lung nodules with shNC and shOTUD3 HCCLM3 cells by tail-vein injection. (G) Images of H&E staining of paraffin-embedded lung tissues from shNC and shOTUD3 nude mice group (Magnification: 100X, 200X, 400X). The unpaired two-sided Student’s t test was used for comparing between two groups of equal variances. Error bars represent mean ± SD from three independent experiments. *P<0.05, **P< 0.01.