Calcitonin
positive regulation of cytosolic calcium ion concentration
Calcitonin promotes outgrowth of trophoblast cells on endometrial epithelial cells: involvement of calcium mobilization and protein kinase C activation.
Embryo implantation is a complex process that requires coordinated trophoblast-endometrial interactions. During implantation, trophoblast cells of the attached blastocyst penetrate the luminal epithelium of the endometrium before invasion into the endometrial stroma. Previous studies demonstrated that calcitonin was actively secreted by rat and human endometrial epithelial cells (EEC) during the implantation window and targeted disruption of endometrial calcitonin expression dramatically decreased embryo implantation rates; however, the role and signal transduction of calcitonin in trophoblast-endometrial interactions remained unclear and are therefore examined in this study. BeWo trophoblast and RL95-2 EEC lines were used because they preserve many properties of their respective normal tissues. We co-cultured BeWo trophoblast spheroids with RL95-2 EEC monolayers to mimic the blastocyst-endometrial interaction, and found that most spheroids quickly attached to EEC monolayers and then progressively expanded, with marked displacement of EEC adjacent to the outgrowing trophoblast cells. Interestingly, pretreatment of EEC monolayers with calcitonin before the addition of spheroids significantly enhanced trophoblast expansion on EEC monolayers. Cytosolic calcium (Ca(2+)) levels in EEC increased rapidly upon exposure to calcitonin, and blockade of Ca(2+) release by BAPTA-AM effectively prevented the promoting effect of calcitonin on trophoblast expansion on EEC. The Ca(2+)-dependent protein kinase C (PKC) was also activated in EEC after calcitonin treatment, and the PKC inhibitors staurosporine and calphostin C could completely abolish calcitonin-induced augmentation of trophoblast expansion on EEC. Our results suggest that calcitonin promotes trophoblastic displacement of EEC through calcium mobilization and PKC activation, thereby facilitating embryo implantation.