In this study, the laboratory-induced strains H128, H256 and H512 obtained from the initial strains of Vibrio parahaemolyticus F7 after continuous in vitro induction with ciprofloxacin were used as the research objects. The biological characteristics of F7, H128, H256, H512 strains, including drug sensitivity, mobility, growth curve and biofilm-forming ability were examined. The relative expressions levels of multidrug efflux efflux pump genes, vmeB, vmeD, vpoC and vmrA were analyzed. The results showed that all the laboratory-induced strains had an increased drug resistance to ciprofloxacin, with their cross-resistance to a variety of non-induced antibiotics also detected. Compared with the original strain F7, the mobility of all the laboratory-induced strains H128, H256 and H512 decreased, with the diameter of moving circle as 4.83 mm, 4.57 mm and 5.13 mm respectively. The biofilm-forming abilities of all laboratory-induced strains were enhanced, with the absorbance reaching 4.35 nm, 4.35 nm and 4.29 nm respectively. The expression level of the efflux pump gene vmeD for all laboratory-induced strains was higher than that of F7 (increase by 1.20, 3.55, and 2.02 times respectively), and the expression levels of the vmeB, vpoC and vmrA genes in H256were higher than that of F7 (increase by 1.47, 2.90, and 1.71 times respectively). These results indicated that the RND efflux system VmeCD-VpoC played important role in the resistance to ciprofloxacin and the main cross-resistance-related efflux systems of Vibrio parahaemolyticus. This study provided a reference for the risk assessment of drug-resistant Vibrio parahaemolyticus, and a new idea for solving the problem of drug resistance of Vibrio parahaemolyticus.