- Tobey Maguire
Peter Parker/Spider-Man - Kirsten Dunst
Mary Jane Watson - James Franco
Harry Osborn - Willem Dafoe
Norman Osborn/Green Goblin - Cliff Robertson
Wujek Ben Parker - Rosemary Harris
Ciocia May Parker - J.K. Simmons
J. Jonah Jameson - Joe Manganiello
Flash Thompson - Bill Nunn
Joseph „Robbie” Robertson - Elizabeth Banks
Betty Brant - Ted Raimi
Hoffman - Ron Perkins
Dr. Mendell Stromm - Gerry Becker
Maximilian Fargas - Jack Betts
Henry Balkan - Stanley Anderson
Generał Slocum - K.K. Dodds
Ellie Simkins - Michael Papajohn
Złodziej samochodu (Dennis Carradine) - Randy Mario Poffo
Bone Saw McGraw - Bruce Campbell
Spiker ringowy