The purpose of this study was to observe whether dual-specificity phosphatase 1 () cells under cold stress. The recombinant plasmids plko.1--shRNA2, plko.1-negative control, and plko.1-EGFP were constructed using the restriction enzymes . These plasmids were separately transduced into 293T cells together with the lentiviral packaging vectors pCMV-DR8.9.1 and pCMV-VSVG. The virus packaged by 293T cells was used to infect the ZF4 cell line derived from zebrafish, and analysis by real-time polymerase chain reaction showed that knockdown ZF4 cell line, the cells were cultured at 28℃ (normal temperature control) or 10℃ (cold stress) for 10 days. Afterwards, they were stained with propidium iodide and annexin V-fluorescein isothiocyanate, then analyzed by flow cytometry. The results showed that the rate of apoptosis of the -knockdown cells increased significantly more under low temperature than that of the wild-type control cells did, as compared with the normal temperature control group. Furthermore, we designed two small hairpin RNAs named kd1 and kd2 based on the coding sequence of . Under cold stress, the frequencies of late and early apoptotic cells in the kd1-treated group were significantly higher, and the frequency of living cells was significantly lower, than those of the negative control group (<0.05). The cells cultured at 28℃ showed no significant difference in the rate of apoptosis according to silencing. However, under cold stress, apoptosis was significantly greater in the is involved in several physiological processes of fish, and it negatively regulates MAPK family proteins; for example, it inhibits p38 phosphorylation. This paper provides a new insight into the regulation of the apoptotic response of fish cells under low-temperature conditions by expression may relieve the inhibition of the MAPK p38 signaling pathway, which promotes ZF4 cell apoptosis under cold stress.