cyclin-d1 and Pre-Eclampsia

Preeclampsia (PE) is a specific disorder of pregnancy with significant morbidity and mortality to the mother and the fetus. It has been reported that abnormal regulation of trophoblast cells contributes to the development of PE. LIM and SH3 Protein 2 (LASP2) belongs to nebulin protein family of actin-binding protein that has broad biological functions. In this study, we evaluated the role of LASP2 in the regulation of trophoblast cells. Our results demonstrated that LASP2 was markedly up-regulated in the placentas of patients with PE. Overexpression of LASP2 significantly suppressed the cell proliferation, migration, and invasion of trophoblast cells. LASP2 overexpression reduced the expression levels of β-catenin, cyclin D1, and c-Myc in trophoblast cells. Furthermore, activation of Wnt/β-catenin pathway booked the effects of LASP2 on trophoblast cells. In conclusion, these findings indicated that LASP2 might contribute to the pathogenesis of PE

Topics: Adult; beta Catenin; Carrier Proteins; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Cytoskeletal Proteins; Female; Humans; LIM Domain Proteins; Placenta; Pre-Eclampsia; Pregnancy; Proto-Oncogene Proteins c-myc; Trophoblasts; Wnt Signaling Pathway

In the present study, we characterized the expression of Activating Protein 1 (AP-1) factors, key cell cycle regulators, in primary placental mesenchymal stromal cells (PDMSCs) derived from normal and preeclamptic (PE) pregnancies with fetal-placental compromise.. PDMSCs were isolated from control (n = 20) and preeclamptic (n = 24) placentae. AP-1 expression was determined by semi-quantitative RT-PCR (sqRT-PCR), Real Time PCR and Western Blot assay. PDMSCs were plated and JunB siRNA was performed. JunB and Cyclin-D1 expression were assessed by Real Time and Western Blot analyses.. JunB expression was significantly increased while Cyclin-D1 expression was significantly down-regulated in PE relative to control PDMSCs. JunB siRNA was accompanied by JunB down-regulation and increased Cyclin-D1 in normal PDMSCs.. We described, for the first time, AP-1 expression in PDMSCs derived from physiological and PE placentae. Importantly, we demonstrated that JunB over-expression in PE-PDMSCs affects Cyclin-D1 regulation. Our data suggest a possible contribution of these pathological placental cells to the altered cell cycle regulation typical of preeclamptic placentae.

Topics: Adult; Case-Control Studies; Cell Cycle; Cyclin D1; Female; Fos-Related Antigen-2; Gene Expression; Humans; Infant, Newborn; Male; Mesenchymal Stem Cells; Placenta; Pre-Eclampsia; Pregnancy; Proto-Oncogene Proteins c-fos; RNA, Messenger; RNA, Small Interfering; Transcription Factor AP-1; Transcription Factors; Young Adult

Preeclampsia (PE) is a pregnancy-specific syndrome manifested by on-set of hypertension and proteinuria after 20 weeks of gestation. Abnormal placenta development has been generally accepted as initial cause of the disorder. Recently, miR-195 was found to be down-regulated in preeclamptic placentas compared with normal pregnant ones, indicating possible association of this small molecule with placental pathology of preeclampsia. By far the function of miR-195 in the development of placenta remains unknown.. Bioinformatic assay predicted ActRIIA as one of the targets for miR-195. By using Real-time PCR, Western blotting and Dual Luciferase Assay, we validated that ActRIIA was the direct target of miR-195 in human trophoblast cells. Transwell insert invasion assay showed that miR-195 could promote cell invasion in trophoblast cell line, HTR8/SVneo cells, and the effect could be abrogated by overexpressed ActRIIA. In preeclamptic placenta tissues, pri-miR-195 and mature miR-195 expressions were down-regulated, whereas ActRIIA level appeared to be increased when compared with that in gestational-week-matched normal placentas.. This is the first report on the function of miR-195 in human placental trophoblast cells which reveals an invasion-promoting effect of the small RNA via repressing ActRIIA. Aberrant expression of miR-195 may contribute to the occurrence of preeclampsia through interfering with Activin/Nodal signaling mediated by ActRIIA in human placenta.

Topics: Activin Receptors, Type II; Adult; Cell Line; Cyclin D1; Cyclin D3; Female; Gene Expression; Gene Expression Regulation; Humans; MicroRNAs; Placenta; Pre-Eclampsia; Pregnancy; Trophoblasts; Young Adult

MiR-155 is known to participate in various cellular processes by targeting gene expression. We previously revealed a link between miR-155 and perturbation of trophoblast invasion and differentiation. This study aimed to investigate the target molecule(s) of miR-155 on the influence on the proliferation and migration of trophoblast cells. Bioinformatics analysis showed that, at the 3' untranslated region (UTR) of cyclin D1, six bases are complementary to the seed region of miR-155. Luciferase assays and cyclin D1 3'UTR transfection assays validated that cyclin D1 3'UTR was the target of miR-155 in HTR-8/SVneo cells. Overexpression of miR-155 in HTR-8/SVneo cells reduced the level of cyclin D1 protein, decreased cell proliferation and invasion, and increased cell number at the G1 stage. Furthermore, the increased expression of miR-155 also regulated the protein levels of kinase inhibitory protein p27 and phosphorylated cytoskeletal protein filamin A. In conclusion, we found that cyclin D1 may be a target of miR-155 in HTR-8/SVneo cells, and demonstrated a negative regulatory role of miR-155 involved in cyclin D1/p27 pathway in proliferation and migration of the cells.

Topics: 3' Untranslated Regions; Adult; Cell Line, Tumor; Cell Movement; Cell Proliferation; Contractile Proteins; Cyclin D1; Down-Regulation; Electron Transport Complex I; Electron Transport Complex III; Electron Transport Complex IV; Female; Filamins; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Microfilament Proteins; MicroRNAs; Pre-Eclampsia; Pregnancy; Trophoblasts

Unexplained intrauterine growth restriction of the fetus (IUGR) results from impaired placental development, frequently associated with maternal malperfusion. Some cases are complicated further by preeclampsia (PE+IUGR). Here, we provide the first evidence that placental protein synthesis inhibition and endoplasmic reticulum (ER) stress play key roles in IUGR pathophysiology. Increased phosphorylation of eukaryotic initiation factor 2alpha suggests suppression of translation initiation in IUGR placentas, with a further increase in PE+IUGR cases. Consequently, AKT levels were reduced at the protein, but not mRNA, level. Additionally, levels of other proteins in the AKT-mammalian target of rapamycin pathway were decreased, and there was associated dephosphorylation of 4E-binding protein 1 and activation of glycogen synthase kinase 3beta. Cyclin D1 and the eukaryotic initiation factor 2B epsilon subunit were also down-regulated, providing additional evidence for this placental phenotype. The central role of AKT signaling in placental growth regulation was confirmed in Akt1 null mice, which display IUGR. In addition, we demonstrated ultrastructural and molecular evidence of ER stress in human IUGR and PE+IUGR placentas, providing a potential mechanism for eukaryotic initiation factor 2alpha phosphorylation. In confirmation, induction of low-grade ER stress in trophoblast-like cell lines reduced cellular proliferation. PE+IUGR placentas showed elevated ER stress with the additional expression of the pro-apoptotic protein C/EBP-homologous protein/growth arrest and DNA damage 153. These findings may account for the increased microparticulate placental debris in the maternal circulation of these cases, leading to endothelial cell activation and impairing placental development.

Topics: Adaptor Proteins, Signal Transducing; Animals; Cell Cycle Proteins; Cell Line; Cell Proliferation; Cyclin D1; Down-Regulation; Endoplasmic Reticulum; Eukaryotic Initiation Factor-2; Female; Fetal Growth Retardation; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Mice; Mice, Knockout; Phosphoproteins; Phosphorylation; Placenta; Pre-Eclampsia; Pregnancy; Protein Biosynthesis; Protein Kinases; Proto-Oncogene Proteins c-akt; TOR Serine-Threonine Kinases; Transcription Factor CHOP