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<i>Listeria monocytogenes</i> is a foodborne pathogen responsible for many food outbreaks worldwide. This study aimed to investigate the single and combined effect of fructooligosaccharides (FOS) and <i>Lactiplantibacillus plantarum</i> subsp. <i>plantarum</i> CICC 6257 (<i>L. plantarum</i>) on the growth, adhesion, invasion, and virulence of gene expressions of <i>Listeria monocytogenes</i> 19112 serotype 4b (<i>L. monocytogenes</i>). Results showed that <i>L. plantarum</i> combined with 2% and 4% (<i>w</i>/<i>v</i>) FOS significantly (<i>p</i> < 0.05) inhibited the growth of <i>L. monocytogenes</i> (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 <i>L. monocytogenes</i> to Caco-2 and BeWo cells were reduced more than 90% compared to the result of the untreated group. After <i>L. plantarum</i> was combined with the 2% and 4% (<i>w</i>/<i>v</i>) FOS treatment, the gene expression of actin-based motility, sigma factor, internalin A, internalin B, positive regulatory factor A, and listeriolysin O significantly (<i>p</i> < 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 <i>L. plantarum</i> and FOS combination against <i>L. monocytogenes</i> was higher than that of FOS or <i>L. plantarum</i> alone. Overall, these results indicated that the <i>L. plantarum</i> and FOS combination might be an effective formula against <i>L. monocytogenes</i>.