The chamber rejected proposals to allow regular scheduled flights to Cuba.
The House also said a rule should remain requiring US citizens to get a special license before going to Cuba.
It was voting on a transport funding bill which has provisions related to Cuba.
President Barack Obama has threatened to veto the bill.
New rules issued by the Obama administration in January were aimed at easing travel restrictions to Cuba and allowing scheduled flights for the first time.
Republican Congressman Mario Diaz-Balart said the White House was wrong to lift the restrictions.
Mario Diaz-Balart, a Cuban-American, said it would mean flights landing at an airport that was partially owned by US interests when it was seized by the Cuban government.
“What you are saying is, <<It’s OK to do business on property that was stolen from Americans>>,” said Mario Diaz-Balart.
The thaw in relations between the US and Cuba was announced at the end of 2014 in simultaneous televised speeches by President Obama and his Cuban counterpart, Raul Castro.
President Barack Obama used his executive powers to ease the embargo that has existed for more than 50 years, defying hardline critics.
However, the issue is caught in a battle between Republicans who control Congress and the White House.
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