Workers' safety was likely put at risk when Construction, Forestry, Maritime, Mining and Energy Union members entered the Transport and Main Roads building in Brisbane on Tuesday morning, Ms Palaszczuk said.
"No worker should go to work fearing for their safety ... it is disgraceful behaviour," she told reporters on Thursday.
"That footage is incredibly disturbing and I would have hated being a person there with that happening ... they owe an apology to those workers who were subjected to that and who felt unsafe."
Ms Palaszczuk said she understood police were looking into the matter.
Police said officers were called to the office building on Mary Street about 9am following reports of a large group entering the premises.
"The group was monitored by officers for several hours but following negotiation a peaceful departure from the group occurred just before 12pm. No further complaints have been made," a spokesperson said in a statement.
A Queensland Transport and Roads Investment Program briefing was cancelled following the action.
"On Tuesday construction workers, frustrated at government inaction over this dysfunctional department, turned up to participate in an industry stakeholder meeting," CFMEU assistant secretary Jade Ingham said in a statement on Thursday.
"Workers - who were at no point asked to leave by police or security - departed of their own accord when the meeting was cancelled by a government too scared to face scrutiny."
Calls for an apology came as Transport Minister Mark Bailey confirmed he met the union on Monday, but had not been told about Tuesday's plans.
Mr Bailey said a late decision not to attend the briefing was due to him having to be at a cabinet budget review committee scheduled at the same time.
"The department knew mid-Monday morning that I couldn't attend the briefing," he said on Thursday.
"There was no indication made to me in the meeting with (CFMEU state secretary Michael Ravbar) on the Monday, he gave no indication about that whatsoever.
"If he had, I would have raised it immediately with my (director-general) and we would have made preparations to make sure that our staff were protected."
Mr Bailey also called on Mr Ravbar to make a public apology.
"Michael Ravbar ... should be making a public apology to the transport and main road road workers who were impacted by their appalling behaviour on Tuesday morning," he said.
"I have backed in our workers from day one as their minister, I take that responsibility seriously, and they were mistreated by his union members."
The opposition is calling for Mr Bailey to be stood down while a full investigation takes place.
"To organise 200 CFMEU members overnight has to be planned, and as if they didn't tell the minister," LNP deputy leader Jarrod Bleijie said on Thursday.
"It's not enough for Mark Bailey just to say I wasn't aware of it. That's rubbish ... he absolutely knew it and the premier has to take action."
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