Fig 1: eIF3b knockdown inhibited cell proliferation, migration, and invasion. A-B: eIF3b knockdown inhibited the proliferation of A498 (A) and CAKI-2 (B) cells. C: EIF3b-depleted cells formed fewer colonies. D: The scratch migration assay indicated that A498 and CAKI-2 migration was inhibited after eIF3b knockdown. E: The transwell assay also revealed that migration was impaired (cell numbers, eIF3b siRNA vs. negative control: A498 cells, 45.3 ± 4.9 vs. 100.3 ± 7.3 [p < 0.01]; CAKI-2 cells, 71.0 ± 6.1 vs. 125.3 ± 7.8 [p < 0.01]), as was the invasive capacity (cell numbers, eIF3b siRNA vs. negative control: A498 cells, 13.3 ± 3.8 vs. 39.7 ± 6.2 [p < 0.05]; CAKI-2 cells, 35.0 ± 6.1 vs. 73.3 ± 8.1 [p < 0.05]), upon eIF3b depletion. F: Changes in cellular morphology after eIF3b depletion.
Fig 2: The animal experiment. A: eIF3b knockdown inhibited the growth of subcutaneous RCC xenografts in mice. B: Xenograft samples after 28 d of growth. C: A498 cells transfected with the eIF3b-RNAi lentivirus (left: eIF3b expression was significantly reduced; right: the transfected cells were abnormal in shape with much more cell debris than control cells) (red arrow: cell debris).
Fig 3: Pathways affected by eIF3b knockdown. A: eIF3b depletion inhibited EMT progression by inactivating the β-catenin pathway. B: The Akt pathway was inhibited after eIF3b depletion.
Fig 4: A: qRT-PCR of fresh samples revealed that eIF3b mRNA expression was downregulated in cancer tissues (p = 0.0137, n = 20) and was significantly greater in metastatic than in non-metastatic tumors (p = 0.0262). B: Kaplan-Meier analysis of the effect of the tumor eIF3b level on OS. C: Kaplan-Meier analysis and log-rank testing of the effect of the TNM grade on OS. D: Kaplan-Meier analysis and log-rank testing of the effect of the nuclear Fuhrman grade on OS. E: Kaplan-Meier analysis of the effect of the ANK eIF3b level on OS. F: The tumor eIF3b expression correlated with the nuclear grade. G: Kaplan-Meier analysis of the effect of the eIF3b level (four categories) on OS. H: Kaplan-Meier analysis of eIF3b level in both tumor and adjacent normal (four categories) on OS.
Fig 5: The effects of eukaryotic translation initiation factor 3 subunit B (EIF3B) knockdown on cell apoptosis and cell cycle. (A) The effects of EIF3B knockdown on cell apoptosis were examined by flow cytometry. (B) The effects of EIF3B knockdown on cell cycle were determined by flow cytometry. Results are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
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