GEN1 regulates cell proliferation, migration, apoptosis and ferroptosis in gastric cancer.

Abstract

Background: Gastric cancer (GC) has a high prevalence and mortality overall. GEN1 is associated with abnormal centrosome amplification, DNA damage and increased apoptosis. To date, little is known about the function and mechanism of GEN1 in GC.

Aim: To explore the cellular processes associated with GC will help to elucidate the mechanism of the occurrence and development of GC and discover potential therapeutic targets.

Methods: The detection of GEN1 expression at mRNA and protein levels was done by real-time quantitative polymerase chain reaction and western blotting. The function of GEN1 was verified by loss-of-function experiments in AGS cells. The genes co-expressed with GEN1 were searched from the stomach adenocarcinomas (STAD) data in The Cancer Genome Atlas database. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the genes co-expressed with GEN1 to further identify the pathways involved in GEN1. Rescue experiments using ferroptosis inhibitor ferrostatin-1 and chemotherapeutic sensitivity assays with cisplatin were also performed.

Results: Significant up-regulation of GEN1 was observed in GC cell lines AGS and MGC-803. Inhibition of GEN1 induced cell apoptosis and decreased cell proliferation, cycle progression, migration in AGS cells. There were 264 genes co-expressed with GEN1 in STAD cohort (r > 0.4, P < 0.001). KEGG enrichment analysis showed that GEN1 might be associated with the cell cycle, Fanconi anemia pathway, homologous recombination, oocyte meiosis and cellular senescence in GC. Furthermore, CCNA2, CCNB1, CCNB2, cyclin-dependent kinase (CDK) 1, CDK2 and polo-like kinase 1 protein levels were lower in GEN1-knockdown AGS cells, manifesting that GEN1 was associated with the cell cycle pathway in AGS cells. Downregulation of GEN1 decreased adenosine triphosphate content and elevated reactive oxygen species in AGS cells, suggesting that GEN1 silencing led to mitochondrial dysfunction in AGS cells. In addition, GEN1 silencing caused an overt decrease in FTH1 and GPX4 protein levels and a significant elevation in ACSL4 protein levels, implying that GEN1 silencing promoted AGS cell ferroptosis. Treatment with ferrostatin-1 rescued cell viability loss induced by GEN1 knockdown, confirming ferroptosis as a key death mechanism. Additionally, GEN1-deficient AGS cells showed enhanced sensitivity to cisplatin, with a significantly reduced half-maximal inhibitory concentration compared to control cells.

Conclusion: GEN1 promotes GC cell proliferation and migration while suppressing apoptosis and ferroptosis. Targeting GEN1 not only disrupts mitochondrial function and cell cycle progression but also sensitizes GC cells to ferroptosis and chemotherapy. These findings highlight GEN1 as a potential therapeutic target for enhancing treatment efficacy in gastric cancer.