Cells of different organs reside in environment of various stiffnesses in vivo. Extracellular matrix (ECM) stiffness has been shown to induce normal cell to have invasive phenotype. In this study, we investigated the effects of ECM stiffness on the phenotypic changes in mechano-sensitive normal keratinocytes in epidermis. After 10days on soft substrates, keratinocytes undergo noticeable changes in the morphology and exhibit mesenchymal features typically observed in fibroblasts. EMT can be characterized by the emergence of vimentin, which is the mesenchymal specific intermediate filament. Real time PCR results show that keratinocyte on soft substrate PDMS increase vimentin mRNA as well as genes related to migration such as RhoA, Cdc42 and Rac1. Inhibition of ROCK showed significant reduction in the responsiveness of keratinocytes to mechanical environment. Our results suggest that soft substrate induces EMT initiation of keratinocyte through ROCK dependent pathways.