Objective: To analyze the effect of transvaginal specimen removal on sexual function, life quality and short-term efficacy of patients diagnosed with colorectal cancer. Method: We retrospectively analyzed the clinical data of 100 patients with colorectal cancer treated in the Cancer Hospital Chinese Academy of Medical Sciences from September 2017 to December 2019. Among them, 52 patients who underwent transvaginal laparoscopic mid-to-high rectal cancer radical resection without assisted abdominal incision who were chosen as the observation group, and 48 patients with conventional laparoscopic-assisted mid-to-high rectal cancer radical resection were set as the control group. Intraoperative blood loss, number of lymph nodes retrieved, operation time, time of urinary catheter removal, drainage tube removal time and postoperative hospital stay were recorded and compared between the two groups. During the process, the Visual Analogue Scale (VAS) was used to assess the pain status of both groups of patients 24 hours after the operation, and the Female Sexual Function Index was used to evaluate the sexual function quality of patients before and 3 months after surgery. Postoperative complications, 2-year survival rate and 6-month postoperative quality of life of both groups of patients were evaluated and compared. Results: In comparison with the control group, the amount of intraoperative blood loss, catheter removal time, drainage tube removal time, and length of postoperative hospital stay were significantly reduced in the observation group, while the number of lymph nodes retrieved was significantly greater. The VAS score 24 hours after operation was lower in the observation group compared with the control group, with statistical significance. There was no significant difference in postoperative sexual function and complication rates between the two groups. However, the 2-year survival rate and the quality of life 6 months after surgery were comparatively higher in the observation group. Conclusion: Natural orifice specimen extraction surgery (NOSES) is effective for transvaginal specimen removal of patients with colorectal cancer, and can significantly improve the prognosis and life quality of patients without affecting their sexual function, with a high safety profile, which is worthy of clinical promotion.