BackgroundSitus inversus totalis (SIT), a rare recessive autosomal disease, involves the complete transposition of the thoracic and abdominal viscera in the left-right axis. Patients with SIT combined with lung cancer are extremely uncommon.Case presentationWe present a case of a 57-year-old woman with SIT who underwent uniportal video-assisted thoracoscopic left middle lobectomy for adenocarcinoma of the lung. The procedure was performed safely with adequate anatomical identification and careful intraoperative manipulation based on the preoperative three-dimensional-computed tomography bronchography and angiography (3D-CTBA). The patient’s perioperative period was uneventful, and no recurrence was observed 2 year postoperatively.ConclusionWith the preoperative planning of the 3D-CTBA, uniportal video-assisted thoracoscopic lobectomy in lung cancer patients with sit can be performed safely and effectively.