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RAD54B promotes gastric cancer cell migration and angiogenesis via the Wnt/β-catenin pathway

Jianchao Li
Jianchao Li
, 
Hui Geng
Hui Geng
, 
Xin Li
Xin Li
, 
Shenshan Zou
Shenshan Zou
 and 
Xintao Xu
Xintao Xu
   | Feb 21, 2024
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Article Category: Research Article
Published Online: Feb 21, 2024
Page range: 67 - 77
Received: Jul 28, 2023
Accepted: Nov 06, 2023
DOI: https://doi.org/10.2478/raon-2024-0007
Keywords
© 2024 Jianchao Li et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

FIGURE 1.

RAD54B expression was increased in gastric cancer. (A) The expression of RAD54B in gastric cancer was enhanced compared to normal samples according to ATGC database. (B) The pan-cancer analysis confirmed an increase in gastric cancer. (C) RAD54B expression was upregulated in gastric cancer based on GEPIA. (D) The expression level of RAD54B was also augmented in gastric cancer cell lines.*p < 0.05; **p < 0.01; ***p < 0.001T = tumor tissues, N = normal tissues
RAD54B expression was increased in gastric cancer. (A) The expression of RAD54B in gastric cancer was enhanced compared to normal samples according to ATGC database. (B) The pan-cancer analysis confirmed an increase in gastric cancer. (C) RAD54B expression was upregulated in gastric cancer based on GEPIA. (D) The expression level of RAD54B was also augmented in gastric cancer cell lines.*p < 0.05; **p < 0.01; ***p < 0.001T = tumor tissues, N = normal tissues

FIGURE 2.

RAD54B facilitated the proliferation of gastric cancer cells. (A) The expression of RAD54B was downregulated or upregulated in AGS and MKN45 cells by the transfection of shRAD54B or overexpression plasmid respectively. (B) Transfected AGS and MKN45 cells were seeded into 6-well plates at a density of 6 × 105 cells per well and maintained at 37 °C for 12 h with 5% CO2. The proliferation of AGS and MKN45 cells was assessed by Edu assays. (C) Transfected AGS and MKN45 cells with 6 × 105 cells per well were plated into 6-well plates and cultured at 37°C for 14 days. The proliferation of AGS and MKN45 cells was determined by colony formation assay.**p < 0.01; ***p < 0.001
RAD54B facilitated the proliferation of gastric cancer cells. (A) The expression of RAD54B was downregulated or upregulated in AGS and MKN45 cells by the transfection of shRAD54B or overexpression plasmid respectively. (B) Transfected AGS and MKN45 cells were seeded into 6-well plates at a density of 6 × 105 cells per well and maintained at 37 °C for 12 h with 5% CO2. The proliferation of AGS and MKN45 cells was assessed by Edu assays. (C) Transfected AGS and MKN45 cells with 6 × 105 cells per well were plated into 6-well plates and cultured at 37°C for 14 days. The proliferation of AGS and MKN45 cells was determined by colony formation assay.**p < 0.01; ***p < 0.001

FIGURE 3.

RAD54B promoted the migration and invasion of gastric cancer cells. The mobility and invasion of AGS and MKN45 cells were assessed by transwell assay using 24-well Transwell chambers with 8.0 μm pore size polycarbonate membranes. 200 μl of cells suspension with a total of 2 × 105 cells was appended into the upper chamber and cultured for 24 h at 37 °C. The migration and invasion of AGS and MKN45 cells were examined by transwell assay.* p < 0.05; **p < 0.01; ***p < 0.001
RAD54B promoted the migration and invasion of gastric cancer cells. The mobility and invasion of AGS and MKN45 cells were assessed by transwell assay using 24-well Transwell chambers with 8.0 μm pore size polycarbonate membranes. 200 μl of cells suspension with a total of 2 × 105 cells was appended into the upper chamber and cultured for 24 h at 37 °C. The migration and invasion of AGS and MKN45 cells were examined by transwell assay.* p < 0.05; **p < 0.01; ***p < 0.001

FIGURE 4.

RAD54B enhanced the tube formation of gastric cancer cells. After HUVECs were inoculated into conditioned medium containing supernatant harvested from transfected AGS and MKN45 cells for co-culture, the tube formation of HUVECs was assessed by tube formation assay.**p < 0.01; ***p < 0.001
RAD54B enhanced the tube formation of gastric cancer cells. After HUVECs were inoculated into conditioned medium containing supernatant harvested from transfected AGS and MKN45 cells for co-culture, the tube formation of HUVECs was assessed by tube formation assay.**p < 0.01; ***p < 0.001

FIGURE 5.

RAD54B enhanced activation of Wnt/β-catenin signaling pathway in gastric cancer cells. The relative protein expressions of β-catenin, Axin, c-myc and MMP-7 were detected by western blot. The data was expressed after being normalized to GAPDH.* p < 0.05; **p < 0.01; ***p < 0.001
RAD54B enhanced activation of Wnt/β-catenin signaling pathway in gastric cancer cells. The relative protein expressions of β-catenin, Axin, c-myc and MMP-7 were detected by western blot. The data was expressed after being normalized to GAPDH.* p < 0.05; **p < 0.01; ***p < 0.001

FIGURE 6.

RAD54B enhanced the growth of gastric cancer and activation of Wnt/β-catenin signaling pathway in vivo. (A) Nude mice were injected with MKN45 transfected with sh-NC or sh-RAD54B#1 and then the tumor volume and weight were monitored for consecutive five weeks after the treatment. (B) The expression levels of RAD54B, Ki-67, c-myc and MMP-7 were determined by IHC.***p < 0.001
RAD54B enhanced the growth of gastric cancer and activation of Wnt/β-catenin signaling pathway in vivo. (A) Nude mice were injected with MKN45 transfected with sh-NC or sh-RAD54B#1 and then the tumor volume and weight were monitored for consecutive five weeks after the treatment. (B) The expression levels of RAD54B, Ki-67, c-myc and MMP-7 were determined by IHC.***p < 0.001
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