De Laurentiis was born at Torre Annunziata in the province of Naples, and grew up selling spaghetti made by his father's pasta factory. He started his studies at the Centro Sperimentale di Cinematografia in Rome in the years 1937–1938 then interrupted by the outbreak of the Second World War.

Following his first movie, L'ultimo Combattimento (1940), Laurentiis produced nearly 150 films during the next seven decades. In 1946 his company, the Dino de Laurentiis Cinematografica, moved into production. In the early years, De Laurentiis produced Italian neorealist films such as Bitter Rice (1949) and the Fellini classics La Strada (1954) and Nights of Cabiria (1956), often in collaboration with Producer Carlo Ponti. In the 1960s, Laurentiis built his own studio facilities, although these financially collapsed during the 1970s. During this period, though, De Laurentiis produced such films as Barabbas (1961), a Christian religious epic; The Bible: In the Beginning (1966), Kiss the Girls and Make Them Die, an imitation James Bond film; Navajo Joe (1966), a spaghetti western; Anzio (1968), a World War II film; Barbarella (1968) and Danger: Diabolik (1968), both successful comic book adaptations; and The Valachi Papers (1972), made to coincide with the popularity of The Godfather.

His brief first marriage in Italy was annulled. In 1949, De Laurentiis married Actress Silvana Mangano, with whom he had four children: Veronica; Raffaella, who is also a film producer; Federico, another Producer who died in a plane crash in 1981 (Dino's movie Dune is dedicated to him); and Francesca. De Laurentiis and Mangano divorced in 1988; she died in 1989. In 1990, he married Martha Schumacher, who produced many of his films since 1985, and with whom he had two daughters, Carolyna and Dina. One of his grandchildren is Giada De Laurentiis, host of Everyday Italian, Behind the Bash, Giada at Home, and Giada's Weekend Getaways on Food Network. He was the younger brother of Luigi De Laurentiis, who became a film Producer after Dino did, and uncle of Aurelio De Laurentiis, also a Producer and the chairman of S.S.C. Napoli football club.

In 1958, he won the Academy Award for Best Foreign Film for producing La Strada, back when producers and Directors would win the award instead of the country it was made in.

De Laurentiis made a number of successful and acclaimed films, including The Scientific Cardplayer (1972), Serpico (1973), Death Wish (1974), Mandingo (1975), Three Days of the Condor (1975), The Shootist (1976), Drum (1976), Ingmar Bergman's The Serpent's Egg (1977), Ragtime (1981), Conan the Barbarian (1982), Blue Velvet (1986) and Breakdown (1997). De Laurentiis' name became well known through the 1976 King Kong remake, which was a commercial hit; Lipstick (1976), a rape and revenge drama; Orca (1977), a killer whale film; The White Buffalo (1977), a western; the disaster movie Hurricane (1979); the remake of Flash Gordon (1980); David Lynch's Dune (1984); and King Kong Lives (1986). De Laurentiis also made several adaptations of Stephen King's works, including The Dead Zone (1983), Cat's Eye (1985), Silver Bullet (1985), and Maximum Overdrive (1986). De Laurentiis's company was involved with the horror sequels Halloween II (1981), Evil Dead II (1987) and Army of Darkness (1992).

De Laurentiis relocated to the US in 1976, and became an American citizen in 1986. In the 1980s he had his own studio, De Laurentiis Entertainment Group (DEG), based in Wilmington, North Carolina. The building of the studio made Wilmington a center of film and television production.

In the 1980s, de Laurentiis owned and operated DDL Foodshow, a specialty retailer with two gourmet Italian markets in New York City and Los Angeles.

De Laurentiis also produced the first Hannibal Lecter film, Manhunter (1986), an adaptation of the Thomas Harris novel Red Dragon. He passed on adapting the novels' sequel, The Silence of the Lambs (1991), but produced the two follow-ups, Hannibal (2001) and Red Dragon (2002), a re-adaptation of the novel. He also produced the prequel Hannibal Rising (2007), which tells the story of how Hannibal becomes a serial killer.

In 2001, he received the Irving G. Thalberg Memorial Award from the Academy of Motion Picture Arts and Sciences.

De Laurentiis died on 10 November 2010 at his residence in Beverly Hills at the age of 91.

In 2012, he received the America Award of the Italy-USA Foundation (in memory).