Objective To evaluate sexual function in women before and after vaginal hysterectomy (VH) and to compare the effects of horizontal and vertical vaginal cuff closure on sexual function. Study Design Women with uterine prolapse of stage 2 or higher were included to this prospective, randomized study. All patients underwent VH with McCall Culdoplasty and patients were randomized into two groups in terms of the vaginal cuff closure technique employed which is either vertically (group 1, right to left) or horizontally (group 2, anterior to posterior). Pelvic Organ Prolapse Urinary Incontinence Sexual Questionnaire—12 Short Form was used to assess sexual function before and 6 months after surgery. Results A total of 78 women participated, 37 in group 1 and 41 in group 2. Significant improvements in were thus evident in both groups 1 (p = 0.000) and 2 (p = 0.000) after surgery; no significant between-group differences were evident. Overall, 61 women (78,2%) had improved PISQ-12 scores postoperatively, 11 (14,1%) scored the same pre- and post-operatively, and 6 (7,9%) scored lower postoperatively. Women who reported poorer sexual function postoperatively, or no improvement, had new-onset or worsening dyspareunia and/or incontinence. Conclusion Most women with uterine prolapse of stage 2 or higher who underwent VH with prolapse repair experienced improved sexual lives postoperatively, regardless of the cuff closure technique used. Although VH to treat POP improves anatomical and sexual concerns, surgery per se may have negative effects on sexual function if new-onset or worsening dyspareunia or incontinence develop.