Blackpool man Glenn Thomas was one of the people killed in the attack.
The protest came on the eve of a trial in the Netherlands of four men - three Russians and a Ukrainian - accused of murder for their alleged involvement.
MH17 was downed by a Buk missile fired from territory held by pro-Russia separatist rebels in 2014.
All 298 people on board died, including 196 Dutch citizens.
Russia has been held politically responsible by the Netherlands and Australia, but denies any involvement.
The chairs were laid out in rows like seats in an airplane and represented those who had died.
White roses were placed on some of the chairs.
Families stood in silence for two minutes and held signs, including one that read: "Impunity = unacceptable! Someone knows what happened ... Justice for MH17."
A placard placed beside the chairs read: "The truth, the whole truth, and nothing but the truth."
Relatives say Russian authorities have attempted to hide what really happened.
Piet Ploeg, whose older brother Alex died in the crash, said families were not accusing Russia of downing the jet.
"But we are having a protest against the lack of cooperation of the Russian state in the investigation of the downing of MH17. And we want them to cooperate and stop obstructing the investigations."
Referring to the chairs set out for victims, he added: "This is for me a very beautiful symbol for 298 people who perished. They are gone. My brother never returned."
Lucas Schansman, whose nephew Quinn died, said: "What we're doing here today, this is truly us making a silent statement to somebody who may have the answers."
Foreign Secretary Dominic Raab said the trial in Amsterdam marks "an important milestone towards accountability for the shooting down of MH17 and the tragic deaths of the 298 people on board".
He added: "The Russian state must now co-operate fully with this trial in accordance with UN Security Council Resolution 2166.
"There can be no impunity for those responsible for this appalling crime."
Last week, a spokesperson for Russian President Vladimir Putin said the Kremlin doubted the objectivity of the Dutch-led investigation.
The four suspects were named last year as Russians Igor Girkin, Sergey Dubinskiy and Oleg Pulatov, and Ukrainian Leonid Kharchenko.
None of them are expected to attend the start of the trial on Monday.
Neither Russia nor Ukraine extradites its citizens.
The trial is expected to last for more than a year.