The Panama Canal Railway (Spanish: Ferrocarril de Panamá) is a railway line linking the Atlantic Ocean to the Pacific Ocean in Central America. The route stretches 47.6 miles (76.6 km) across the Isthmus of Panama from Colón (Atlantic) to Balboa (Pacific, near Panama City). Because of the difficult physical conditions of the route and state of technology, the construction was renowned as an international engineering achievement, one that cost US$8 million and the lives of an estimated 5,000 to 10,000 workers. Opened in 1855, the railway preceded the Panama Canal by half a century; the railway was vital in assisting the construction of the canal in the early 1900s. With the opening of the canal, the railroad's route was changed as a result of the creation of Gatun Lake, which flooded part of the original route. Following World War II, the railroad's importance declined and much of it fell into a state of neglect until 1998, when a project to rebuild the railroad to haul intermodal traffic began; the new railroad opened in 2001.

The line was built by the United States and the principal incentive was the vast increase in passenger and freight traffic from eastern USA to California following the 1849 California Gold Rush. The United States Congress had provided subsidies to companies to operate mail and passenger steamships on the coasts, and supported some funds for construction of the railroad, which began in 1850; the first revenue train ran over the full length on January 28, 1855. Referred to as an inter-oceanic railroad when it opened, it was later also described by some as representing a "transcontinental" railroad, despite traversing only the narrow isthmus connecting the North and South American continents. For a time the Panama Railroad also owned and operated ocean-going ships that provided mail and passenger service to a few major US East Coast and West Coast cities, respectively.

Known as the Panama Railroad Company when founded in the 19th century, today it is operated as Panama Canal Railway Company (reporting mark: PCRC). Since 1998 it has been jointly owned by Kansas City Southern and Mi-Jack Products and leased to the government of Panama. The Panama Canal Railway is primarily dedicated to freight transport, but it also operates a passenger service between Panama City and Colón.