Cold drawing pearlitic steel wires with ultra-high strength have important applications such as the automobile tire, springs rope and bridge cables. There have been many investigations which are paid attention to the hardening mechanisms of pearlitic steel wire, covering evolution of microstructure, texture and dislocation. In this work, effects of pearlitic lamella orientation on mechanical properties and deformation of pearlite steel wire in cold drawing were investigated by combining TEM, SEM and nano-indentor. The experimental results showed that pearlitic lamellae having low angle with drawing direction would be turned to parallel the drawing direction through a combined rotating process of pearlite colonies. The <110> fiber texture in the ferrite phase formed and the distribution of dislocations in ferrite was almost uniform. The deformation between cementite and ferrite was coordinated. Pearlitic lamellae having large angle with drawing direction would be bent and turned to parallel the drawing direction. It was difficult for the bent pearlite to deform because of the fragmentation of cementite and formation of dislocation cell in ferrite. After cold drawing process, the micro-hardness of straight pearlite was higher than that of bent pearlite. The result indicated that pearlitic lamellae having low angle with drawing direction have higher working-hardening rate in drawing.