In the relapsed patients, 3 (6.8%) were local recurrences and 2 (4.5%) were distant metastases. There was no significant difference between clinicopathological factors and recurrence; however, patients with Ki-67 LI less than 5% showed better disease-free survival than patients with Ki-67 LI over 5% (p = 0.04). In multivariate Cox regression analysis, although there was no significantly different in disease-free survival by age, histology, tumor size, only Ki-67 LI showed a significant prognostic factor of recurrence (HR = 12.5, 95% CI = [1.1-1407], p = 0.04). Conclusions: Ki-67 LI after VATS segmentectomy was a prognostic factor of disease-free survival in NSCLC and the treatment of choice for patients with positive LI may be considered, in addition to adjuvant chemotherapy, or lobectomy.