The Queen serves as the primary antagonist in "Hello Kitty in Alice in Wonderland," ruling Wonderland alongside the King of Hearts with an impulsive, tyrannical nature. She habitually demands immediate beheadings for perceived slights, shouting "Off with their heads!" when angered or disobeyed.

Her wrath first falls upon anthropomorphic cards caught secretly painting white roses red to evade punishment. She condemns them to execution, but Alice intervenes, enabling their escape and drawing the Queen's fury onto herself.

The Queen then forces Alice into a chaotic croquet match. This game employs live flamingos as mallets and hedgehogs as balls, descending into disarray due to the Queen's unfair rules and the creatures' erratic movements. Alice's struggle to control the game further incenses the Queen.

Later, presiding over a trial accusing the Jack of Hearts of stealing her cherry pie, the Queen faces Alice's defense. Alice challenges the Mad Hatter's testimony and exposes the Queen's injustice, prompting the Queen to shift her focus and demand Alice's execution. This confrontation escalates as Alice grows in size, disrupts the courtroom, and escapes Wonderland, causing the Queen's subjects to flee. Her depiction remains consistent as an authoritarian antagonist enforcing obedience through fear within this specific adaptation.