- Lindsay Lohan as Cady Heron, a 16-year-old girl who transfers to a public high school after being homeschooled her whole life in Africa
- Rachel McAdams as Regina George, a rich, popular mean girl and the leader of The Plastics
- Tina Fey as Ms. Norbury, math teacher and Mathletes moderator
- Tim Meadows as Mr. Duvall, the principal of North Shore High School
- Amy Poehler as Mrs. George, Regina's mother
- Ana Gasteyer as Mrs. Heron, Cady's mother
- Lacey Chabert as Gretchen Wieners, a rich Jewish girl and member of the Plastics, Regina's second-in-command
- Lizzy Caplan as Janis Ian, Cady and Damian's goth friend and artist
- Daniel Franzese as Damian, Cady and Janis' openly gay friend
- Neil Flynn as Mr. Heron, Cady's father
- Jonathan Bennett as Aaron Samuels, Regina's ex-boyfriend and Cady's love interest
- Amanda Seyfried as Karen Smith, a dimwitted member of the Plastics
- Rajiv Surendra as Kevin Gnapoor, the Mathletes president and an amateur rapper who is attracted to Janis
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