Commissioner for Equality, Helena Dalli, said: "EU values and fundamental rights must be respected by Member States and state authorities."
The European Union has rejected grants under a twinning programme to six Polish cities because of their attitude to the LGBTQ community.
The announcement was confirmed on Twitter by the EU's Equality Commissioner, Helena Dalli.
"EU values and fundamental rights must be respected by member states and public authorities," Helena Dalli said on Tuesday.
“This is why six town twinning applications involving Polish authorities that adopted ‘LGBTI free zones’ or ‘family rights’ resolutions were rejected.”
It is unclear which towns have not been selected for grants.
The cities and municipalities had applied for a grant between €5,000 and €25,000 under the twinning programme of the Europe for Citizens project, which aims to stimulate debate and civic participation on EU policies.
Applications for 127 cities and projects — eight Polish beneficiaries — were selected by the EU for a total grant value of more than €2.3 million.
"The call for proposals for the twinning programme stipulates that it must be accessible to all European citizens without any form of discrimination on the basis of gender, ethnic origin, religion or belief, disability, age or sexual orientation," a Commission spokesman told AFP News Agency on Wednesday.
Those applications that were rejected on the basis of not being in line with the programme's objectives of "equal access and non-discrimination".
Poland's Minister of Justice, Zbigniew Ziobro, has condemned the EU's decision, saying his country will defend the rule of law.
"We will not allow discrimination of Polish citizens and local governments by the European Union," Ziobro said on Facebook.
"The Union must respect the equality of all its citizens, who have the right to form their opinions and beliefs freely."
In 2019, 80 municipalities in Poland declared themselves "free of LGBT ideology", supported by local politicians associated with the ruling Conservative Law and Justice party.
The European Parliament condemned the move from Poland and called for increased monitoring of the use of EU funds.
Meanwhile, in February, French councillors in Saint-Jean-de-Braye announced that they would be ending "official relations" with the town of Tuchów over anti-LGBTQ sentiments.
"France is committed to combating human rights violations based on sexual orientation," the council said in a statement.
"We cannot accept that the ties that unite our two cities through an oath of twinning should be tainted.
"We reaffirm that human rights must apply without discrimination to every human being, regardless of sexual orientation or gender identity."