assessment of the effect of overexpression of death-associated protein kinases 3 using pegfpn1 on gastric adenocarcinoma cell line (mkn45)

Background: gastric cancer (gc) is one of the most common malignancies worldwide. an in-depth understanding of the molecular mechanisms that underlies tumor gc will lead to breakthroughs in the targeted treatment of gc. based on multiple lines of evidence, death-associated protein kinase 3 (dapk3) regulates both programmed cell death including apoptosis and autophagy. the widespread experimental evidence raises the possibility of using dapk-based gene therapy strategies. objectives: the aim of this study was to investigate the effect of overexpression of dapk3 using the pegfpn1 vector on the gastric adenocarcinoma cell line (mkn45). methods: the mkn45 cell lines were cultured in a dmem culture medium and, then, the recombinant vector pegfpn1-dapk3 was transfected into the cells by lipofectamine 2000. the effects of the overexpression of the dapk3 gene on mkn45 cells were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2h-tetrazolium bromide (mtt), flow cytometry, and real-time quantitative reverse transcription pcr (qrt-pcr) techniques. results: our findings indicated that overexpression of dapk3 in mkn45 cells not only affects the expression of apoptosis-related genes but also changes the expression of autophagy-related genes. additionally, overexpression of dapk3 reduces the metabolic activity of cells. conclusions: the overexpression of the dapk3 gene can lead to cell death by both inducing apoptosis and autophagy pathways in the gastric adenocarcinoma cell line (mkn45). this anti-cancer activity may describe a hopeful strategy in the application of novel gene therapy for the treatment of gastric adenocarcinoma; however, further research is required to examine the clinical effective ness of this strategy in gc treatment.