She was born Virginia Cathryn Rowlands in Cambria, Wisconsin, she attended the University of Wisconsin. Her father, Edwin Myrwyn Rowlands, was a state Assemblyman and her mother, Mary Allen Neal, was a housewife.

Rowlands appeared in Broadway in the late 1950s, and made her film debut in The High Cost of Loving in 1958. She starred in several anthology television series, including Robert Montgomery Presents, Kraft Television Theatre and Studio One, among many others. In 1961 she starred in the well-received television series 87th Precinct, and in 1964 in Peyton Place (TV series).

Teaming with her husband, writer and director John Cassavetes, Rowlands starred in many productions, including Staccato, A Child Is Waiting, Faces, Gloria (nomination for Academy Award for Best Actress), Love Streams, Minnie and Moskowitz, She's So Lovely, and A Woman Under the Influence (Academy Award nomination). She starred in The Neon Bible. In 1985, Rowlands played the mother in the critically acclaimed made-for-TV movie An Early Frost. In recent years, she has appeared in Paulie and in Mira Nair's HBO movie, Hysterical Blindness for which she won her third Emmy.

She was recently seen in The Notebook, which was directed by her son, Nick Cassavetes, and co-starred James Garner, Ryan Gosling and Rachel McAdams. In 2004 she won her first Daytime Emmy for her role as Mrs. Evelyn Ritchie in The Incredible Mrs. Ritchie. To name a few, Ms. Rowlands has been nominated for: two Academy Awards; six Emmy nominations, and one Daytime Emmy; eight Golden Globes; three Satellite Awards; and one SAG Award. Some of her notable wins include: a Silver Berlin Bear; three Emmy Awards and one Daytime Emmy; two Golden Globes; two National Board of Review Awards; two Satellite Awards; and one Prize San Sebastián.

In 2005, she appeared opposite Kate Hudson, Peter Sarsgaard, and John Hurt in the gothic thriller The Skeleton Key.