【Objective】 To explore the effects of estradiol-17 beta and estrone on gap junction intercellular communication and Cx43 and Cx32 expression of stromal and glandular cells in vitro. 【Methods】 The scrap loading dye transfer was used to investigate the effects of E1 and E2 on GJIC of endometrial stromal and glandular cells. The influence of E1 and E2 on Cx43 and Cx32 expressions was investigated with a laser scanning confocal microscope. 【Results】 The ability of GJIC function was higher in the endometrial stromal cells than that of the glandular cells in vitro. 2.5 × 10-5 mol/L E1 leading to smaller GJIC after 48 h(P < 0.05). Cx32 expression in endometrial stromal and glandular cells were significantly higher than Cx43. 5 × 10-5 mol/L E2 and 2.5 × 10-5 mol/L E1 had no effect on Cx expression. 【Conclusions】 The function of Estrone’s down-regulation on GJIC of endometrial stromal and glandular cells may be one of mechanisms which invert endometrial hyperplasia. E1 and E2 had no effect on Cx expression of endometrial stromal and glandular cells, which indicate Cx expression could not completely reflect GJIC function in vitro.