The c-MYC proto-oncogene encodes a ubiquitous transcription factor involved in cell proliferation and tumorigenesis. Heat shock protein 60 (HSP60) plays an essential role in assisting many newly synthesized proteins to reach their native forms. Increased HSP60 expression is observed in different types of human cancer. Here we show that c-MYC directly activates HSP60 transcription through an E-box (CACGTG) site located in the proximal promoter of the HSP60 gene. Overexpression of HSP60 induces transformation. Short-interference RNA (siRNA) mediated repression of HSP60 reduces transformation caused by c-MYC overexpression. These results indicate that c-MYC may promote transformation through the induction of HSP60 expression.