Hefner was born in Chicago, Illinois. She is the daughter of Mildred (Williams) and Hugh Hefner. Her parents had separated by the time she was five. When her mother remarried, she moved to Wilmette, Illinois. There she graduated from New Trier High School. She attended the National Music Camp at Interlochen during the summers from 1964-1969.
She graduated summa cum laude from Brandeis University with a bachelor's degree in English and American literature in 1974. She was elected to Phi Beta Kappa in her junior year.
After college, she free-lanced for the "Boston Phoenix" for a year, writing movie reviews. A year later, she moved back to Chicago and started working at Playboy. After four years, she was promoted to vice president.
In 1982, she became president of Playboy Enterprises, and was made chairman of the board and CEO in 1988. The company acquired adult-oriented businesses such as Spice Network and ClubJenna.
In 2008, she released a memo to employees about her efforts to streamline the company's operations, including eliminating its DVD division and laying off staff.
On December 8, 2008, she announced her plans to step down as CEO of Playboy as of January 31, 2009. Hefner said that the election of Barack Obama as the next U.S. president had inspired her to give more time to charitable work, and that the decision to step down was her own. "Just as this country is embracing change in the form of new leadership, I have decided that now is the time to make changes in my own life as well," she said.
In May 2011, she was named executive chairman of Canyon Ranch Enterprises, a resort company that operates six premier spa destinations and an online website providing health and wellness advice.
When she left Canyon Ranch Enterprises in 2015, she became Chairman of Hatch Beauty, a brand and product innovation company in the personal care space, and she also joined the board of RDO Companies, a $3 billion family owned agricultural conglomerate.
She's a founding member of The Chicago Network, The Committee of 200, and Women Corporate Directors. She's also an advisory board member to Springboard Enterprises and a Rush University Medical Center trustee.
Hefner created the Hugh M. Hefner First Amendment Award in honor of her father, and has helped to raise $30 million to build the CORE Center in Chicago, the first outpatient facility in the Midwest for people with AIDS.
She married former Illinois state senator William A. Marovitz, a real estate developer and attorney in 1995, and she lives in Chicago. They divorced in 2013, and they have no children. Marovitz was sued by the Securities Exchange Commission, in 2012, for allegedly using inside information to trade illegally in shares of Playboy. He settled out of court for $168,352.