Background and objective Video-assisted thoracoscopic surgery (VATS) lobectomy is generally accepted for patients with lung cancer. The aim of this study is to explore the feasibility of the single-operation-hole thoracoscopic lobectomy in the treatment of non-small cell lung cancer. Methods To review and analyze the single-operation-hole thoracoscopic lobectomy performed in our hospital for 113 non-small cell lung cancer (NSCLC) cases from October 2010 to October 2013. The incision for observation was 1.5 cm the eighth intercostal at the rear of the midaxillary line and the incision for operation was 2.0 cm-4.0 cm at the fourth or fifth intercostal of the anterior axillary line. The operations were performed through the single-operation-hole. Result The operation processes were smooth for all the patients without any operative mortality occurrence. Only in 5 cases was the operation hole expanded because of the occurrence of massive hemorrhage during the operation; 3 patients with postoperative complications underwent thoracoscopic lobectomy again, including 2 cases of delayed hemorrhage and 1 case of chylothorax. The average surgical duration was (178.24±31.17) min, the average blood loss was (213.56±62.38) mL, and the number of lymph nodes dissected was from 5-22. All diagnose were confirmed by pathology after operation. The average length of stay was (8.17±2.93) d. All cases recovered well during the follow-up of (2-38) months, only 5 cases had recurrence or metastasis. Conclusion The single-operation-hole thomcoscopic lobectomy for lung cancer is safe and feasible, further reducing the trauma, and can be used as a conventional treatment for early- or medium-term NSCLC..