Previous studies have reported that E2F transcription factor 1 (E2F1) and DNA topoisomerase II alpha (TOP2A) are up-regulated in gastric cancer (GC), the corresponding unexplored regulatory mechanisms and the interactions of which in GC are worth figuring out. In this study, the expressions of E2F1 and TOP2A in GC tissues were assessed by real-time PCR (RT-PCR), and E2F1 binding sites in the TOP2A promoter region were predicted via bioinformatics analyses and confirmed using dual-luciferase reporter and ChIP assays. The viability, apoptosis, migration, and invasion of GC cells after transfection with sh-E2F1 or TOP2A plasmid were measured using methyl thiazolyl tetrazolium (MTT) assay, Hoechst 33258 staining/Annexin V/PI staining, and transwell assay. The expressions of E2F1, TOP2A, Cleaved caspase-3, Proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin, and Vimentin in transfected cells were assessed by reverse transcription quantitative PCR (RT-qPCR) and western blot. Concretely, E2F1, as a transcription factor, has binding sites in the TOP2A promoter region. E2F1 and TOP2A levels were up-regulated and positively correlated with each other in GC tissues, i.e., overexpressed E2F1 increased TOP2A levels and E2F1 silencing decreased TOP2A levels in GC cells. Moreover, E2F1 silencing hindered GC cell viability, migration, and invasion, enhanced apoptosis, diminished the levels of PCNA, N-cadherin and Vimentin, and elevated those of Cleaved caspase-3 and E-cadherin in GC cells. However, overexpressed TOP2A reversed the above effects of E2F1 silencing. To sum up, E2F1-mediated up-regulation of TOP2A promotes the viability, migration, and invasion, and inhibits the apoptosis of GC cells.