Journal of Southern Medical University ›› 2023, Vol. 43 ›› Issue (4): 537-543.doi: 10.12122/j.issn.1673-4254.2023.04.05

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Overexpression of miR-431-5p impairs mitochondrial function and induces apoptosis in gastric cancer cells via the Bax/Bcl-2/caspase3 pathway

WU Jiaming, DENG Zhongquan, ZHU Yi, DOU Guangjian, LI Jin, HUANG Liyong   

  1. Department of Gastrointestinal Surgery, First Hospital of Jiaxing (First Affiliated Hospital of Jiaxing University), Jiaxing 314000, China
  • Online:2023-04-20 Published:2023-05-16

Abstract: Objective To investigate the expression of microRNA miR-431-5p in gastric cancer (GC) tissues and its effects on apoptosis and mitochondrial function in GC cells. Methods The expression level of miR-431-5p in 50 clinical samples of GC tissues and paired adjacent tissues was detected using real-time fluorescence quantitative PCR, and its correlation with the clinicopathological features of the patients was analyzed. A cultured human GC cell line (MKN-45 cells) were transfected with a miR-431-5p mimic or a negative control sequence, and the cell proliferation, apoptosis, mitochondrial number, mitochondrial potential, mitochondrial permeability transition pore (mPTP), reactive oxygen species (ROS) production and adenosine triphosphate (ATP) content were detected using CCK- 8 assay, flow cytometry, fluorescent probe label, or ATP detection kit. The changes in the expression levels of the apoptotic proteins in the cells were detected with Western blotting. Results The expression level of miR-431-5p was significantly lower in GC tissues than in the adjacent tissues (P<0.001) and was significantly correlated with tumor differentiation (P=0.0227), T stage (P=0.0184), N stage (P=0.0005), TNM stage (P=0.0414) and vascular invasion (P=0.0107). In MKN-45 cells, overexpression of miR-431-5p obviously inhibited cell proliferation and induced cell apoptosis, causing also mitochondrial function impairment as shown by reduced mitochondrial number, lowered mitochondrial potential, increased mPTP opening, increased ROS production and reduced ATP content. Overexpression of miR-431-5p significantly downregulated the expression of Bcl-2 and increased the expressions of pro-apoptotic proteins p53, Bcl-2 and cleaved caspase-3 protein. Conclusion The expression of miR-431-5p is down-regulated in GC, which results in mitochondrial function impairment and promotes cell apoptosis by activating the Bax/Bcl-2/caspase 3 signaling pathway, suggesting the potential role of miR-431-5p in targeted therapy for GC.

Key words: gastric cancer; cell apoptosis; mitochondrial function; miRNA-431-5p