Inhibition of gastric cancer cell proliferation and invasion by circ-BCAR3 through regulation of miR-145-5p/VEGFR-2 signaling pathway
10.3760/cma.j.cn121382-20241013-00609
- VernacularTitle:circ-BCAR3通过调控 miR-145-5p/VEGFR-2信号通路对胃癌细胞增殖和侵袭的抑制作用
- Author:
Yanjuan XIONG
1
;
Yong LIU
;
Huiqin PENG
;
Yong XU
;
Ting GUO
Author Information
1. 黄石市中心医院（湖北理工学院附属医院）消化内科，黄石 435000
- Keywords:
Gastric cancer;
Circ-BCAR3;
MiR-145-5p;
Cell proliferation;
Cell invasion
- From:
International Journal of Biomedical Engineering
2024;47(6):577-583
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze the expression and significance of circ-BCAR3 in gastric cancer tissues, and to explore the effect and molecular mechanism of silencing circ-BCAR3 on the proliferation and invasion of gastric cancer cells. Methods:The expression of circ-BCAR3 in gastric cancer tissues and adjacent normal tissues was analyzed using the UCSC database. The relationship between circ-BCAR3 expression and the survival rate of gastric cancer patients was analyzed by Kaplan-Meier method. Small interfering RNA (siRNA)-negative control (si-NC) and si-circ-BCAR3 were transfected into gastric cancer BGC823 cells, and recorded as si-circ-BCAR3 group and si-NC group, respectively. Real-time reverse transcription-PCR (RT-qPCR) was used to detect the expression of circ-BCAR3 gene in gastric cancer NCI-N87, AGS, BGC823, HS-746T, SGC7901 cells, and immortalized gastric mucosal epithelial GES-1 cells. The proliferation and invasion abilities of BGC823 cells were detected by clone formation assay and Transwell assay, respectively. The binding sites of circ-BCAR3 and miR-145-5p were predicted using the online Circinteractome platform, and their targeting relationship was verified by a dual luciferase reporter gene assay. The relationship between the expression of circ-BCAR3 and miR-145-5p in gastric cancer tissues was analyzed using the UCSC database. The expression levels of vascular endothelial growth factor receptor-2 (VEGFR-2) signaling pathway proteins, including phosphorylated VEGFR-2 (p-VEGFR-2), phosphorylated phospholipase C-λ (p-PLC-λ), phosphorylated extracellular signal-regulated kinase (p-Erk), and phosphorylated P38 (p-P38), were detected in BGC823 cells using Western blotting analysis. Results:The relative expression level of circ-BCAR3 in gastric cancer tissues (8.04±2.34) was significantly higher than that in adjacent normal tissues (2.53±1.74), and the difference was statistically significant ( P<0.01). The survival rate of gastric cancer patients with low circ-BCAR3 expression was higher than that of gastric cancer patients with high circ-BCAR3 expression, and the difference was statistically significant ( P<0.05). The relative expression levels of circ-BCAR3 in NCI-N87, HS-746T, AGS, BGC823, and SGC7901 cells (5.05±0.36, 3.50±0.21, 2.87±0.38, 6.80±0.29, and 3.77±0.53) were all higher than those in GES-1 cells (1.02±0.21), and the differences were statistically significant (all P<0.01). The number of clones formed in the si-NC group and si-circ-BCAR3 group were [(126.70±21.78) and (44.89±8.33) unit], respectively, and the number of invasive cells were [(53.81±5.92) and (24.91±4.56) unit], respectively, with statistically significant differences ( P<0.01). The AACUGGA sequence in circ-BCAR3 was found to have the capacity to bind to the UUGACCU sequence in miR-145-5p. The luciferase activity of wild type circ-BCAR3 (WT- circ-BCAR3)+ miR-145-5p group (0.26±0.03) was lower than that of WT- circ-BCAR3+miR-NC group (1.00±0.05), and the difference was statistically significant ( P<0.01). In contrast, for the miR-NC, the luciferase activities of the mutant- circ-BCAR3 (MUT- circ-BCAR3)+ miR-145-5p and MUT- circ-BCAR3+ miR-NC groups were (1.07±0.12) and (1.00±0.09), and the differences were not statistically significant ( P>0.05). The expression of circ-BCAR3 and miR-145-5p showed a negative correlation in gastric cancer tissues ( R=0.82, P<0.01). The relative expression levels of VEGFR-2 signaling pathway proteins p-VEGFR-2, p-PLC-λ, p-Erk, and p-P38 in BGC823 cells in the si-circ-BCAR3 group (1.08±0.21, 1.50±0.21, 3.01±0.25, and 0.46±0.10) were lower than those in the si-NC group (5.22±0.17, 5.56±0.19, 6.63±0.31, and 4.24±0.16), and the differences were statistically significant (all P<0.05). Conclusions:The expression of circ-BCAR3 is elevated in gastric cancer tissues and cells. Furthermore, silencing of circ-BCAR3 may inhibit the proliferation and invasion of BGC823 cells by regulating the miR-145-5p/VEGFR-2 signaling pathway.
