Havana and Washington have been at odds since the Cuban revolution, led by Fidel Castro, toppled Fulgencio Batista’s regime in 1959.
The United States imposed a partial trade embargo on the Caribbean island nation in October 1960.
Washington imposed a full trade embargo on Cuba in February 1962.
The Republic of Cuba says the United States diplomatic mission in Havana is conducting subversive activities devised to destabilize the government of President Raul Castro.
"The US Interests Section in Cuba continues to serve as a general headquarters for the subversive policies of the North American government," the Cuban Foreign Ministry said in a statement issued on Friday, The Associated Press reported.
The United States and Cuba have not had formal diplomatic ties since 1961. However, Washington maintains an interests section on the island.
The Cuban Foreign Ministry said US officials unlawfully give classes inside the walls of the interests section and provide Internet service to anti-Castro politicians without the government’s permission.
It added that Havana would defend Cuba's sovereignty "by any legal means" at its disposal.
The Cuban Foreign Ministry also stated that the US Interests Section was attempting to accomplish "the impossible task of converting its mercenaries into a credible internal opposition movement."