Introduction: Natural orifice specimen extraction (NOSE) laparoscopic surgery is emerging as a new and promising technique, especially in gastric cancer (GC). Aim: To analyze the difference between transvaginal and transumbilical specimen extraction following totally laparoscopic gastrectomy (TLG)Material and methods: Between January 2016 and July 2021, 37 consecutive female patients with GC who underwent either natural orifice specimen extraction surgery (NOSES) or TLG at our center were included and analyzed. Results: In total, 12 patients underwent NOSES, and 25 patients underwent TLG. The NOSES group had a shorter operative time (239.3 ±21.5 vs. 256.1 ±21.2 min, p = 0.031) and shorter specimen extraction time (17.0 ±4.2 vs. 30.8 ±4.3 min, p < 0.01). No significant difference was observed in the comparison of the radical validity including estimated blood loss, the number of harvested LNs and the comparisons of distal and proximal margin. In the postoperative recovery comparisons, the NOSES group had a shorter time to first fluids (3.9 ±0.5 vs. 5.6 ±1.2 days, p < 0.01), and time to starting a soft diet (5.6 ±0.7 vs. 7.7 ±1.7 days, p < 0.01). Postoperative pain in the NOSES group was significantly less. Postoperative hospital stay days in the NOSES group were fewer (10.2 ±2.2 vs. 12.4 ±2.9 days, p = 0.030). Overall, the postoperative complications were similar between the two groups (p = 0.438). When more cases were included, the results were similar. NOSES reduced hospital costs for GC patients, and it also applied to the premenopausal patients. Conclusions: NOSES was a safer and more reliable method as compared to transumbilical specimen extraction.