|
|
|
| The expression of cyclin D1 of endometrial stromal cells regulated by IKKα and its decidualization |
| Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, 430000 |
|
|
|
Abstract To study the change of cyclin D1 expression of endometrial stromal cells by down-regulating IKKα, and investigate its influences on the process of endometrial decidualization and the embryo implantation. In vitro decidualization of hESCs was induced by 8-Br-cAMP and IKKαsiRNA were transfected into hESC vitro cultured. While the cells in negtive control group was transfected by NC siRNA. In vitro decidualization of the hESC was induced by 8-Br-cAMP and MPA, and the morphology of cells was observed. The mRNA and protein expressions of IKKα, cyclin D1, prolactin (PRL), and insulin like growth factor binding protein 1 (IGFBP-1) of the cells in the two groups were measured by realtime-PCR and Western blotting. The cell cycle was tested by flow cytometer. The mice model with pseudopregnancy endometrium decidualization was constructed, and IKKα siRNA was injected in one side of uterus cornu of the mice, while NC siRNA was injected in other one side of uterine cornu as the negative control. The changes of endometrial stromal cell morphology and glandular ducts in bilateral uterine cornu of the mice were observed after HE staining. The expression of IGFBP-1 was measured by immunohistochemistry. Realtime PCR and Western blotting was used for detecting the expressions of IKKα,cyclin D1, PRL, and IGFBP-1. IKKα(CHUK)/ NC siRNA was injected into the uterus cornu of the sexually mature female kunming mice after feed together with the normal male kunming mice. The embryo implantation sites in bilateral uteri of the mice were recorded. Results: 8-Br-cAMP and MPA can induce hESCs decidualization in vitro. The levels of PRL mRNA (339.00±52.25 fold), IGFBP-1 mRNA
(199.6±9.36 fold), and IGFBP-1 protein (4.86±0.30) of the mice endometrial cells in decidual intervention group were significant higher than those of the mice endometrial cells in the control group. There was a blocking from G0-1 phase to S phase of hESCs of the cells. Downregulation of IKKα in vitro leaded to the overexpression of Cyclin D1 and thus hindered the decidualization of hESCs in vitro. There was a blocking from G0-1 phase to S phase of hESCs of the cells in the control group. Injection of castor oil into the uterine cornu of pseudopregnant mice had induced the decidualization of endometrium. Down-regulated IKKα expression of cells in mice could affect endometrial decidualization and embryo implantation. Conclusion: Down regulation of IKKαexpression of endomentrial stromal cells may lead to overexpression of cyclin D1, and then inhibit the decidualization process in vivo and in vitro, which will affect embryo implantation.
|
|
|
|
|
|
|
|
|
