1. Many clinical studies have suggested a relationship between oestrogen and insulin sensitivity. In the present study, HepG2 cells were divided into four groups: (i) control, incubated with 1 nmol/L insulin; (ii) the HI group, which was incubated with 100 nmol/L insulin to induce insulin resistance; (iii) the E2 group, in which control cells were incubated with 1 nmol/L insulin plus 1 nmol/L oestradiol; and (iv) the HI + E2 group, in which insulin-resistant cells were incubated with 100 nmol/L insulin + 1 nmol/L oestradiol. 2. A high concentration of insulin decreased the activity of phosphofructo-1-kinase (PFK), pyruvate dehydrogenase (PDH) and glycogen synthase (GS), as well as decreasing the expression of insulin receptor (IR) and insulin receptor substrate-2 (IRS-2). High insulin had no effect on glucose transport or the expression of insulin receptor-1 (IRS-1). 3. The addition of oestradiol to control cells increased glucose transport, the activity of PFK, PDH and GS and the expression of IRS-1 and IRS-2, but had no effect on the expression of IR. 4. Treatment of insulin-resistant HepG2 cells with oestradiol attenuated HI-induced decreases, except for IR, and the expression of IRS-1 was significantly higher than control, attaining levels seen in group 3. The expression of IRS-2 was significant higher than in insulin-resistant cells, but did not reach control levels. Changes in the activity of PFK, PDH and GS were the same as the changes seen in the expression of IRS-2. 5. These results suggest that high concentrations of insulin induce insulin resistance in HepG2 cells, whereas oestradiol improves glucose metabolism and insulin signal transduction of cells by enhancing the activity of key enzymes involved in glucose metabolism and the expression of IRS-1 and IRS-2.