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The objective of this study is to investigate the relationship between caspase-3 expression and apoptosis in the gastric adenocarcinoma cell line SGC-7901 transfected with the adenovirus-mediated FasL gene. The human gastric adenocarcinoma cell line SGC-7901 was transfected with the adenovirus-mediated FasL gene. The expression of caspase-3 and apoptotic morphological changes were measured by flow cytometry, TUNEL, RT-PCR, and immunocytochemical assays. The expression of caspase-3 was low in the gastric adenocarcinoma cell line SGC-7901, but transfection of these cells with Ad-FasL enhanced the cellular expression of the caspase-3 gene and induced susceptibility to Fas/FasL-mediated apoptosis (25.4% vs. 7.2%, p < 0.001). Apoptosis exhibited a positive relationship with caspase-3 expression (gamma = 0.647, p < 0.01). The down-regulation of caspase-3 expression and resistance to Fas/FasL-mediated apoptosis in gastric adenocarcinoma cells were found to be related to the development and progression of the cells. FasL gene transfer may increase caspase-3 expression in gastric adenocarcinoma cells and enhance apoptosis in them.