Situs ambiguous is a rare anomaly, which includes various abnormalities of position and development of trunk organs and results in diagnostic and therapeutic problems during major abdominal intervention. We report the case of a woman with peripheral papillary cholangiocarcinoma and hilar extension, developed on situs ambiguous associated with the following abnormalities: agenesis of the retrohepatic vena cava, preduodenal portal vein, a variant of the hepatic arteries, truncated pancreas, polysplenia, and mesenteric malrotation. After complete anatomical assessment, resection of segments 4 to 8 extended to the common bile duct with lymphadenectomy and reconstruction by hepaticojejunostomy was performed with no surgical complications. The patient was alive with no signs of recurrence at 18 month follow-up. The specificities of situs ambiguous must be identified by anatomical assessment but do not prevent complex abdominal surgery. We report the case of a woman with peripheral papillary cholangiocarcinoma and hilar extension, developed on situs ambiguous associated with the following abnormalities: agenesis of the retrohepatic vena cava, preduodenal portal vein, a variant of the hepatic arteries, truncated pancreas, polysplenia, and mesenteric malrotation. After complete anatomical assessment, resection of segments 4 to 8 extended to the common bile duct with lymphadenectomy and reconstruction by hepaticojejunostomy was performed with no surgical complications. The patient was alive with no signs of recurrence at 18 month follow-up. The specificities of situs ambiguous must be identified by anatomical assessment but do not prevent complex abdominal surgery.