Objective To inquire into the influence of the pelvic floor reconstruction surgery of pelvic organ prolapse on Papillomavirus-Negative atypical squamous cells of undetermined significance cytology. Methods This investigation was studied retrospectively about female patients of POP 3~4 stage with high-risk human papillomavirus DNA-negative ASCUS between January, 2008 and September, 2018.All patients received one of two treatment regimens, pelvic floor reconstruction or local hydrocortisone cream(0.1% Hydrocortisone Butyrate cream, 1g nightly for no less than 12 weeks). After 6 months follow-up, we collected data to compare the positive rate of ASCUS with high-risk human papillomavirus DNA-negative between the two groups after intervention. Results The negative conversion rate of ASCUS in the operative patients was higher than in patients using local hydrocortisone cream (33/47 [70.2%] vs 19/44 [43.2%], P = 0.009). Conclusions The decline of non–human papillomavirus ASCUS cytology in postoperative patients might be related to pelvic floor reconstruction.