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Some <i>Latiactobacilli</i> are often used as probiotics due to their functional activities, including antioxidant, anticancer and immunoregulation effect. <i>Loigolactobacillus coryniformis</i> NA-3 obtained from our laboratory is a promising probiotic according to the previous study. Coculture, the Oxford cup test and disk-diffusion methods were used to evaluate the probiotic properties and antibiotic resistance of <i>L. coryniformis</i> NA-3. The antioxidant activities of live and heat-killed <i>L. coryniformis</i> NA-3 were assessed via radicals’ scavenging ability. The potential anticancer and immunoregulatory capacity was determined in vitro using cell lines. The results indicate that <i>L. coryniformis</i> NA-3 has antibacterial activity and cholesterol removal ability and is sensitive to most antibiotics. Dead <i>L. coryniformis</i> NA-3 can scavenge free radicals as well as live strains. Live <i>L. coryniformis</i> NA-3 can significantly inhibit the proliferation of colon cancer cells; however, dead cells cannot. After RAW 264.7 macrophages were treated with live and heat-killed <i>L. coryniformis</i> NA-3, the production of NO, IL-6, TNF-α and reactive oxygen species (ROS) was induced. The increased expression of inducible nitric oxide synthase (iNOS) in treated macrophages mediates the production of NO. In conclusion, <i>L. coryniformis</i> NA-3 showed potential probiotic properties, and the heat-killed strain also exhibited activities similar to those of live bacteria, suggesting the possible value of its further application in the food processing and pharmaceutical industries.