The Worth Street station was constructed for the Interborough Rapid Transit Company (IRT) as part of the city's first subway line, which was approved in 1900.

Construction of the line segment that includes the Worth Street station started on July 10 of the same year.

The station was closed on September 1, 1962, as a result of a platform lengthening project at Brooklyn Bridge–City Hall.

[4]: 21  However, development of what would become the city's first subway line did not start until 1894, when the New York State Legislature passed the Rapid Transit Act.

It called for a subway line from New York City Hall in lower Manhattan to the Upper West Side, where two branches would lead north into the Bronx.

[4]: 161  The Rapid Transit Construction Company, organized by John B. McDonald and funded by August Belmont Jr., signed the initial Contract 1 with the Rapid Transit Commission in February 1900,[6] in which it would construct the subway and maintain a 50-year operating lease from the opening of the line.

[5]: 4  Belmont incorporated the Interborough Rapid Transit Company (IRT) in April 1902 to operate the subway.

[2][4]: 186 The first collision in the history of the New York City Subway system occurred at the station on January 11, 1905.

East Side local trains ran from City Hall to Lenox Avenue (145th Street).

[13] On March 20, 1906, the IRT tested a vacuum cleaner connected to a portable wagon plant to clean the station.

[14][15] To address overcrowding, in 1909, the New York Public Service Commission proposed lengthening the platforms at stations along the original IRT subway.

[16]: 168  As part of a modification to the IRT's construction contracts made on January 18, 1910, the company was to lengthen station platforms to accommodate ten-car express and six-car local trains.

[17]: 15  The northbound platform of the Worth Street station was extended about 10 feet (3.0 m) northward into the "electric manhole", a passageway leading to the equipment closet.

[23][24] The downtown platform was lengthened in 1948 by the New York City Board of Transportation, providing for the full length of a ten-car, 514-foot-long (157 m) train.

If the station were retained, service on the line would be slowed down, and there was no suitable signal system that could operate with such a short distance.

[37] Additional columns between the tracks, spaced every 5 feet (1.5 m), support the jack-arched concrete station roofs.

[5]: 4 [32]: 9  Alternating columns between the local and express tracks had black on white signs reading "Worth.