Objective: To investigate the effects of inhibiting ARTN gene by targeted interference of ARTN gene on proliferation of endometrial cancer Ishikawa cells. Methods: Short hairpin RNA (pGPU6/GFP/Neo-ARTN shRNA) interference plasmid of ARTN gene was constructed using pGPU6/GFP/Neo as carrier, and transfected into human endometrial cancer Ishikawa cells by liposome method. The transfection efficiency was observe under fluorescence microscope; Western Blot was used to detected ARTN protein expression in Ishikawa cells after transfection; CCK method to detect the proliferative activity of Ishikawa cells. Results: The pGPU6/GFP/Neo-ARTN shRNA interference plasmid was successfully transfected into Ishikawa cells and the transfection efficiency was 80%. 48h after transfection, the ARTN protein expression of Ishikawa cells in interference group was obviously lower than negative control group and normal group (P<0.05). 48h, 72h and 96h after transfection, the proliferative activity of Ishikawa cells in interference group was obviously lower than negative control group and normal group (P<0.05). Conclusions: Targeted interference of ARTN gene can inhibit the proliferative activity of endometrial cancer Ishikawa cells.