Echoing the "Death to America" phrase chanted throughout the 1979 unrest, student protesters on Monday gathered outside the former embassy building in a repeated show of anti-American sentiment.
Protesters were seen carrying placards saying "down with USA", and a cardboard cutout of US President Donald Trump. Some were also seen burning American flags.
According to state media, another 1,000 demonstrations were held across the country.
It comes also as Iranian politicians gave initial approval for a bill that would seek to include information about "America's crimes" in school textbooks.
In a speech outside the former embassy on Monday, army chief Major General Abdolrahim Mousavi said: "Our fight with America is over our independence, over not submitting to bullying, over values, beliefs and our religion.
"They (Americans) will continue their hostilities, like the proverbial poisonous scorpion whose nature it is to sting and cannot be stopped unless it is crushed."
On November 4, 1979, students stormed the embassy building in the Iranian capital and took 52 US nationals hostage.
They were freed 444 days later after the inauguration of US president Ronald Reagan.
This action on the embassy which was replayed in the 2012 movie Argo came as part of the followup from Shah Mohammad Reza Pahlavi fleeing Iran in January of that year in a movement eventually known as the Islamic Revolution.
But it was him seeking treatment for terminal cancer in New York that insensed protesters involved in unrest and power struggeles back home, and led to the siezure of the embassy.
However, Iran's marking of the anniversary was somewhat overshadowed by an attack on its consulate in neighbouring Iraq, during protests on Sunday night that left three Iraqis dead.
Amid the demonstration, protesters in the city of Karbala were said to have climbed the building and hung banners to demand Iran stay out of Iraqi politics.
It also comes as Iran prepares to announce its plans to pull further away from its promises in the 2015 nuclear treaty in response to the US backing out last year.
