In vivo and in vitro studies on apoptosis in OSE cells and inclusion cysts of pregnant heifers.

Elevated progesterone concentration during pregnancy and use of progesterone-like contraceptives are known to reduce ovarian cancers. This study was undertaken to decipher whether or not there is any relationship between progesterone (also oestrogen)-mediated ovarian surface epithelium (OSE) apoptosis and expression of p53, a cell-cycle arresting protein and potential tumour suppressor. Immunohistochemical staining with cytokeratin confirmed epithelial nature of the cells in the OSE layer and inclusion cysts that invaginate inside stroma after ovulation takes place. The in situ apoptosis index was determined during oestrus, and at mid and late-pregnancy stages in heifers. Epithelia of both tissues exhibited significantly high nuclear staining, suggesting that these cells are aiming to apoptotic destruction. To further establish a role of progesterone, the OSE cells were exposed in vitro to two concentrations of oestrogen and progesterone. It was revealed that progesterone at both concentrations and oestrogen only at high concentration converted a large proportion of these cells apoptotic. The stimulatory effect of progesterone (and to some extent oestrogen) was also seen on p53 expression in the same cultivated OSE cells. The steroid dosage dependence for apoptosis and p53 expression was also somewhat similar. Assuming that progesterone action is mediated through p53-caused apoptosis as a mechanism to evade malignant transformation of OSE cells, p53 expression at mRNA and protein level was investigated in the OSE layer in proximity to stroma, antrum and corpus luteum (CL). In cycling animals CL produces a large amount of progesterone and also oestrogen to maintain the post-ovulatory cycle and to suppress the gonadotropin production. Hence, cells undergoing re-epithelialization and which are in contact with CL were expected to undergo maximum apoptotic modification. Indeed we got the maximum p53/p53 gene expression in these cells. We conclude that progesterone during cycling and pregnancy may reduce the risk of developing ovarian cancer by ceasing cell cycle and diverting damaged and mutagenized OSE cells for apoptosis, and the process may be mediated through elevated p53 synthesis. However, it is also possible that progesterone and p53-induced apoptosis may be entirely different cancer suppressive actions but coincidently happening together.