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After studying theatre at the University of the Philippines Diliman, De Leon began acting on stage. Her earliest performances include local productions of The Merchant of Venice, Waiting for Godot, Old Times, and Medea. She made her film debut in Peque Gallaga's horror anthology Shake, Rattle & Roll III (1991) and was cast in small and uncredited roles throughout the 1990s and 2000s. De Leon made brief appearances in films and guest-starred on television shows. Her breakthrough came in the crime drama Verdict (2019), for which she won a FAMAS Award for Best Supporting Actress. In 2021, she portrayed an ageing sex worker in Lav Diaz's period drama History of Ha to positive reviews. She followed it with more assertive parts in Erik Matti's HBO television projects On the Job and Folklore.

De Leon achieved international recognition and acclaim for her performance as a toilet cleaner on a luxury yacht in Ruben Östlund's satirical black comedy Triangle of Sadness (2022), winning the Guldbagge Award and the Los Angeles Film Critics Association Award for Best Supporting Performance. She received nominations for a Golden Globe Award and a BAFTA Award for Best Supporting Actress, becoming the first Filipino to be nominated in any category. De Leon has since starred in the adult animated drama The Missing (2023)—for which she won a Cinemalaya Award for Best Supporting Actress—and the 2024 comedy dramas Between the Temples and Ghostlight. She is the co-founder of Ladies Who Launch, a social services group which supports disadvantaged communities.