K416B Ni-based superalloy with high W content has good high temperature properties and low cost, which has a great development potential. To investigate the room temperature tensile property and the deformation feature of K416B superalloy, tensile testing at room temperature was carried out, and optical microscopy (OM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to analyze the deformation and damage mechanisms. Results show that the main room temperature tensile deformation features of the K416B nickel-based superalloy are dislocations slipping in the matrix and shearing into γ' phase. The super-dislocations shearing into γ' phase can form the anti-phase boundary two coupled (a/2) partial-dislocations or decompose into the configuration of two (a/3) partial dislocations plus stacking fault. In the later stage of tensile testing, the slip-lines with different orientations are activated in the grain, causing the stress concentration in the regions of block carbide or the porosity, and cracks initiate and propagate along these regions.