Han Hye-jin debuted in the 2001 short film Magic Thumping. In 2004, she was cast in TV Novel: You Are a Star, in which she won the Best New Actress award at the KBS Drama Awards. In 2005, she landed her breakthrough role as a young widow who dreams of becoming a successful hairdresser in the hit daily MBC drama Be Strong, Geum-soon! Her performance in the drama earned her a Top Excellence Award at the 2005 MBC Drama Awards. In 2006, she became part of the sageuk blockbuster Jumong. The drama was slated to air for 60 episodes but was extended to 81 episodes due to its immense popularity. Han Hye-jin won a Top Excellence Award at the MBC Drama Awards and also scored a Best Actress nomination at the 43rd Baeksang Arts Awards for her work in the sageuk drama. In 2011, she took home the Popularity Award at the KBS Drama Awards for the romantic melodrama The Thorn Birds.

Aside from acting, Han Hye-jin also hosted the talk show Healing Camp from 2011 to 2013. She earned various citations for her work in the program, including an Excellence Award at the 2012 SBS Entertainment Awards.

Han Hye-jin was in a relationship with Brown Eyed Soul member Naul from 2003 to 2012. On July 1, 2013, she married South Korean professional footballer Ki Sung-yong. Their daughter was born on September 13, 2015.