Abstract： Objective To examine the effect of sex hormones on the growth and gene expression of human papillomavirus (HPV) type 16 DNA-immortalized human uterine cervical epithelial cells (HCE16/3 cells).Methods The effect of sex hormones on the growth and viral gene expression of HCE16/3 cells was studied using [3 H]-thymidine incorporation, soft agarose assay and Northern blot analysis.Results The growth of HCE16/3 cells was found to be little affected by estradiol and progesterone while insulin was a mitogen for HCE16/3 cells in phenol red-free medium with steroid-stripped serum. Furthermore, synergistic effects between insulin and hormones were not observed. Estradiol could not induce the growth of HCE16/3 cell line in soft agarose, either. In Northern blot analysis, however, the hormones upregulated the HPV 16 early gene expression in HCE16/3 cells, which was generally considered to be required for the proliferation of HPV DNA-immortalized cells.Conclusions These results suggest that the proliferation of HCE16/3 cells is still dependent on growth factors and sex hormones upregulate the HPV 16 early gene expression. More researches should be done before elucidating the effects of sex hormones on human cervical cancinogenesis.