"The course of events taking place in Ukraine now, completely out of line with the 1998 treaty on friendship, cooperation and partnership, is unacceptable," Luzhkov said at a session of Moscow's alternative "student government" during a discussion of state policy regarding compatriots abroad.
"I am convinced our country should make the decision to end this treaty instead of extending it," Luzhkov said, adding that the treaty in question failed because of the Ukrainian authorities' attitude.
The Moscow mayor said Russian language teachers in Crimea receive smaller wages than other subject teachers.
"This is state policy of Ukrainian authorities to oust the Russian language, when all the Crimea and part of Ukraine think in Russian and speak it," he said.
Russian-language programming has disappeared from Ukrainian state television channels in recent years and the education system has been switched to Ukrainian, even though Russian is the native language of many people in eastern Ukraine and Crimea.
Frequent disputes have flared up between Russia and Ukraine over the lease of naval facilities on the Crimean peninsula. In the latest row, Luzhkov was barred from entering the former Soviet republic after a speech in which he questioned Ukraine's right to sovereignty over the region.
Russia's Black Sea Fleet currently uses the Sevastopol base under an agreement signed in 1997. Ukrainian President Viktor Yushchenko recently decided not to extend the lease beyond May 28, 2017.
In early June, Russia's lower house of parliament adopted a resolution saying the Russian-Ukrainian cooperation treaty could be denounced if Ukraine joins NATO.
Ukraine's pro-Western leadership has been pursuing NATO membership since 2004, when President Viktor Yushchenko came to power. Ukraine failed to secure an agreement on a NATO Membership Action Plan, a key step toward joining the alliance, at the organization's summit in April, but was told the decision would be reviewed in December.