Listeria monocytogenes is a foodborne pathogen responsible for many food outbreaks worldwide. This study aimed to investigate the single and combined effect of fructooligosaccharides (FOS) and Lactiplantibacillus plantarum subsp. plantarum CICC 6257 (L. plantarum) on the growth, adhesion, invasion, and virulence of gene expressions of Listeria monocytogenes 19112 serotype 4b (L. monocytogenes). Results showed that L. plantarum combined with 2% and 4% (w/v) FOS significantly (p < 0.05) inhibited the growth of L. monocytogenes (3–3.5 log10 CFU/mL reduction) at the incubation temperature of 10 °C and 25 °C. Under the same combination condition, the invasion rates of L. monocytogenes to Caco-2 and BeWo cells were reduced more than 90% compared to the result of the untreated group. After L. plantarum was combined with the 2% and 4% (w/v) FOS treatment, the gene expression of actin-based motility, sigma factor, internalin A, internalin B, positive regulatory factor A, and listeriolysin O significantly (p < 0.05) were reduced over 91%, 77%, 92%, 89%, 79%, and 79% compared to the result of the untreated group, respectively. The inhibition level of the L. plantarum and FOS combination against L. monocytogenes was higher than that of FOS or L. plantarum alone. Overall, these results indicated that the L. plantarum and FOS combination might be an effective formula against L. monocytogenes.