Madhu Shalini was born and brought up in Hyderabad, Andhra Pradesh, her father being a businessman and her mother an advocate. Like her mother she also learned Kuchipudi, before taking part in a beauty contest, which she incidentally won, which paved her the way for a modelling career. After working as a television anchor for a short while, she soon ventured into film business, turning an actress.
She made her feature film debut with the film, E. V. V. Satyanarayana's Kithakithalu opposite Allari Naresh, released in 2006, went to become a very successful venture. She had appeared in two more Telugu films, namely Teja's Oka V Chitram, and Aganthakudu in the same year. She made her Tamil film debut later that year with the low-budget feature Pazhaniappa Kalloori, She later signed to play the lead female in D. Sabhapathy's Telugu project Happy Journey, while she also agreed to appear in the director's next Tamil venture, Pathinaaru. Though the film received mixed reviews and fared averagely at the box office. Her subsequent release, Karalu Miriyalu, likewise, released to negative reviews, while her performance; however, met with favorable remarks from critics, she eventually got her big break as she was cast by noted National Film Award-winning filmmaker Bala for his comedy flick Avan Ivan. She made her Bollywood debut in Ram Gopal Varma's Department that featured her as a gun moll alongside Amitabh Bachchan, Sanjay Dutt and Rana Daggubati. Madhu Shalini will be seen as a reporter in Gopala Gopala lastly she was seen in Thoongavanam Tamil movie and in Seethavalokanam.