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[Vitamin D induces proliferation in rat endometrium cultured cells]
F Delissalde1, M A Hernández, A Barrón
1Departamento de Bioquímica y Biología Molecular, Instituto Nacional de Perinatología, México D.F.
Objective:
To study the effects of 1,25-dihydroxyvitaminD3 (1,25-(OH)2D3) on proliferation and cell death in the rat uterus.
Material And Methods:
A rat endometrial cell line (Rentro 1) grown in a Dulbecco Minimal Essential Medium (DMEM) supplemented with 1% charcoal stripped serum was used in all experiments in order to eliminate the steroid hormone. Cell monolayer was incubated in the presence and absence of 1,25-(OH)2D3 or 17 beta-estradiol or vehicle. After stimulation, we evaluated cell proliferation and DNA synthesis by trypan blue counting method and flow cytofluorometry, respectively. Finally, the genomic DNA integrity was evaluated by electrophoresis and the bands visualized with ultraviolet light.
Results:
The cells in medium containing 1% fetal bovine serum free of steroid hormones stimulated the cell growth 85% more than without serum. Supplement with albumin did not allow cell growth. The cells did not respond to 17 beta-estradiol but the presence of 1,25-(OH)2D3 induced cell proliferation. These results confirm that Rentro 1 cells do not express the estrogen receptor and demonstrate their capacity to respond to 1,25-(OH)2D3. Finally, the integrity of DNA was not affected by 1,25(OH)2D3, suggesting that this hormone is not involved in cell death by apoptosis in our cell line, as seen in other cell lines.
Conclusions:
1) 1,25-dihydroxyvitamin D induced cell proliferation in the endometrial cell line Rentro 1 in a dose-dependent fashion and this effect is independent of the presence of an estrogenic stimulus; 2) the increase in cell number was related to DNA synthesis during the cell cycle; and 3) the presence of the hormone in the culture medium was not able to induce cell death.