Objective To investigate the pathological roles of postnatally up-regulated alpha-fetoprotein(AFP) expression in ZBTB20 knockout mice in postnatal growth retardation, metabolic dysfunction, and pre-mature mortality. Methods ZBTB20 and AFP knockout mice were generated by crossbreeding ZBTB20 knockout mice with AFP knockout mice.The postnatal growth, survival rate, and blood glucose of ZBTB20/AFP double knockout mice were observed and compared with those of ZBTB20 knockout mice. Results There were no significant differences in postnatal growth, survival, and glucose homeostasis between ZBTB20/AFP doube knockout mice and ZBTB20 knockout mice. Conclusion The postnatally up-regulated liver AFP in ZBTB20 knockout mice is not associated with the growth retardation, hypoglycemia, or pre-mature mortality.