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Summary: To explore the role of insulin and forkhead box protein O1 (FoxO1) on cell proliferation regulated by sirtuin 1 (Sirt1) in goose primary hepatocyte, goose primary hepatocytes were isolated and incubated with Sirt1 inhibitor and nicotinamide, and then, some hepatocytes were transfected with FoxO1 interference plasmid or treated with insulin. The indexes related with cell proliferation were measured. The results first showed Sirt1 inhibition induced by nicotinamide increased cell proliferation in goose primary hepatocytes (P < 0.05). Second, the cyclin D1 protein content of cotreatment of nicotinamide and miRNA-FoxO1 was higher than that of nicotinamide treatment (P < 0.05); rate of bromodeoxyuridine-positive cell of the cotreatment with nicotinamide and FoxO1 interference was between the single nicotinamide treatment and the single FoxO1 interference treatment (P < 0.05); there was no significant difference between nicotinamide treatment and cotreatment of nicotinamide and miRNA-FoxO1 in DNA synthesis rate (P > 0.05), protein content of p21 (P > 0.05), and cell proliferative index (PI) (67.40 vs. 69.00%). Finally, the PI of cotreatment with nicotinamide and insulin (56.65%) was lower than the single nicotinamide treatment (67.40%); the DNA synthesis rate in cotreatment of nicotinamide and insulin was lower than the single nicotinamide treatment (P < 0.05). These findings suggested that Sirt1 inhibition increased cell proliferation in goose primary hepatocytes, FoxO1 and insulin may play role in cell proliferation mediated by Sirt1 in goose primary hepatocyte; different influence is only observational, and further investigations are require to highlight the potential role of FoxO1 and insulin in cell proliferation mediated by Sirt1 in goose primary hepatocyte.