The effects of laser treatment scanning patterns on the properties of Ti-V-Hf-Zr non-evaporable getter (NEG) film on 316L stainless steel (316L SS) substrates were analyzed. 316L SS substrates were laser-treated with different patterns, which resulted in spherical structure and protrusions on the surface of the substrates. Subsequently, Ti-V-Hf-Zr NEG films were deposited on ultrasonically cleaned laser-treated 316L SS by a direct current (DC) magnetron sputtering machine. Based on the XPS results of Ti-V-Hf-Zr, it has been revealed that the atomic proportions of film elements on the surface of samples #1-1∼#3–1 were about 2.2 (Ti):1.7 (Zr):0.9 (V):1.0 (Hf). In this paper, the Ar+-induced secondary electron yield (SEY) and surface resistivity of Ti-V-Hf-Zr NEG are discussed for the first time, and it has been observed that the laser treatment leads to an increase of the substrate surface roughness, which decreases the SEY and increases the surface resistivity.