The PI3K inhibitors LY294002 and wortmannin, and the MEK ERK inhibitors U0126 and PD98059

MMP 26, also known as endometase or matrilysin JAK 阻害剤 2, was recently identified as the smallest member of MMP family. It is widely expressed in villous CTBs, syn cytiotrophoblasts and EVTs of human placenta. MMP 26 exhibits wide substrate specificity in cleaving ECM and basement membrane proteins including type IV collagen, fibronectin, fibrinogen, vitronectin and gela tin. The fact that most of these ECM components are expressed in human villous trophoblasts suggests the involvement of MMP 26 in the degradation and remodelling of ECM at the feto maternal interface. In addition, MMP 26 is able to process the lacent MMP 9, to generate its active form. Our previous studies revealed that the spatiotemporal expression of MMP 26 was similar to that of MMP 9 in trophoblasts during the first trimester, implicating that MMP 26 may also indirectly contribute to ECM degradation through acti vation of proMMP 9 at the feto maternal interface.<br><br> Recently, we found that overexpression of MMP 26 increases invasive capacity of human trophoblast cells buy LDE225 in vitro, suggesting the important role of MMP 26 in trophoblast invasion. However, little is known about the regulatory mechanism of MMP 26 expression in human trophoblasts. An increasing body of evidence has shown the extrapi tuitary functions of gonadotropin releasing hormone I and the second mammalian form of this hor mone. In human placenta, GnRH I is widely expressed among distinct subpopulations of trophoblasts throughout gestation, while GnRH II expression is restricted to mononuclear villous CTBs and EVTs during the first trimester.<br><br> GnRH receptor is highly expressed in both CTBs and syncytiotropho blasts during early gestation. Recently, GnRH I and GnRH II have also been shown to regulate the expres sion of MMP 2, MMP 9tissue inhibitor of metallopro teinases 1, and urokinase plasminogen activator plasminogen activator inhibitor in human EVT cultures. These observations LY2109761 ic50 suggest the paracrine andor autocrine regulatory roles of these two hormones in modulating the activities of various protease systems at the feto maternal interface. Based on previous findings from our laboratory and others, we hypothesized that GnRH I and GnRH II induce MMP 26 expression in trophoblast cells. In the present study, we investigated the regulatory mechanism of MMP 26 expression by GnRH I and GnRH II using an in vitro experimental model of an immortalized human cytotrophoblast like cell line, B6Tert 1, which has been established in our laboratory.<br><br> Methods Materials GnRH I and GnRH II native peptides were obtained from Peninsula Laboratories, Inc. ERK12 inhibitor and JNK inhibitor were purchased from Sigma Aldrich Corp. PD98059 and SP600125 were dissolved in DMSO. The antibodies specific against b actin, phospho ERK12, phospho JNK, total ERK12 and total JNK were purchased from Cell Signaling Technology, Inc. The antibodies specific against GnRH receptor were obtained from Lab Vision Corp. The polyclonal antibodies against MMP 26 are kind gifts from Dr. Qing Xiang A. Sang of Florida State University, Florida, USA. Cell culture and treatment Immortalized human cytotrophoblast like B6Tert 1 cells were cultured as described previously.