ß-TrCP suppresses the migration and invasion of trophoblast cells in preeclampsia by down-regulating Snail.

Insufficient trophoblast invasion has been shown to contribute to the occurrence and progression of preeclampsia (PE). Recently, beta-transducin repeat containing E3 ubiquitin protein (ß-TrCP) was shown to function as a ubiquitination regulator in regulating the proliferation and invasion of various cell types. In this study, we employed an in vitro model of trophoblasts to investigate the role played by ß-TrCP in the pathogenesis of PE. The levels of ß-TrCP in newly delivered placentas from 15 pregnant women with PE and 15 healthy pregnant women were detected by quantitative real-time PCR and western blot assays. The effects of ß-TrCP on cell migration, invasion, and epithelial-mesenchymal transition (EMT) in two trophoblast cell lines (HTR-8/SVneo and TEV-1) were examined using wound healing assays, Transwell assays, and western blot assays, respectively. Rescue experiments were performed by treating ß-TrCP knockdown or ß-TrCP expressing trophoblasts with si-Snail transfection or a proteasome inhibitor (MG132). ß-TrCP mRNA and protein expression levels were significantly increased in the PE placentas when compared to the normal control placentas. ß-TrCP overexpression significantly inhibited cell migration and invasion, while silencing of ß-TrCP promoted cell migration and invasion of the two trophoblast cell lines. Furthermore, we demonstrated that ß-TrCP-mediated ubiquitination might inhibit the EMT process of trophoblasts by down-regulating Snail expression. Our results suggest that both ß-TrCP mRNA and protein expression were up-regulated in the PE placentas. ß-TrCP impeded the migration and invasion of trophoblasts by suppressing Snail expression. This implicates the ubiquitin-proteasome pathway in the pathogenesis of PE, and suggests ß-TrCP as a potential target for treating PE.

[Analysis of normal pelvis morphometry of modern Chinese southern Han female and its correlation with age].

OBJECTIVE: To analyze the normal pelvis morphometry of Chinese southern Han female and its correlation with age. METHODS: From August 2009 to September 2011, 289 Han nationality females who received pelvis CT scan at Nanfang Hospital of Southern Medical University were eligible for the study. Their mean age was 43.5 years, with normal body development and no pelvic abnormality. The patients were divided into 3 age groups: 25 to 40 (n = 109), 41 to 50 (n = 115), and >50 years (n = 65). After constructing a three-dimensional digital model of the pelvis, the following parameters were measured, including transverse inlet diameter, posterior sagittal diameter of pelvic inlet, sagittal inlet, diagonal conjugate, biischial diameter, posterior sagittal diameter of midpelvis, sagittal midpelvic diameter, intertuberous distance, posterior sagittal diameter of outlet, angle of pubic arch, sagittal outlet, penal height, sacrum length and sacrum curvature. The relationship between all parameters and age was analyzed. RESULTS: (1) The posterior sagittal diameters of pelvic inlet of the 25 to 40 age group, 41 to 50 age group, >50 years age group were (53 ± 8), (51 ± 7), (48 ± 6) mm. The sagittal inlet of the three groups were (122 ± 8), (120 ± 9), (114 ± 8) mm. And the diagonal conjugate of the three groups were (135 ± 10), (132 ± 9), (127 ± 9) mm. All had significant differences among the three groups (P < 0.01) . (2) The posterior sagittal diameter of midpelvis of the three groups were (43 ± 6), (44 ± 6), (43 ± 7) mm, and the sagittal midpelvic diameter of the three groups were (119 ± 8), (120 ± 8), (119 ± 7) mm, with no significant difference among the three groups (P > 0.05). (3) The intertuberous distance of the three groups were (122 ± 11), (121 ± 10), (117 ± 11) mm, and the posterior sagittal diameter of outlet of the three groups were (56 ± 9), (58 ± 8), (57 ± 9) mm. There was no significant difference among the three groups (P > 0.05) .(4) Penal height of three groups were (31 ± 3), (33 ± 3), (34 ± 3) mm, there was with significant differences (P < 0.01) .(5) Pearson correlation analysis showed that penal height was positively correlated with age (r = 0.331, P < 0.05), while sagittal inlet, posterior sagittal diameter of inlet, diagonal conjugate, intertuberous distance, angle of pubic arch, sacrum length and sacrum curvature were negatively correlated with age (r = -0.120 to -0.343, P < 0.05). CONCLUSIONS: The shape of the pelvic inlet of Chinese southern Han female changed from sagittal elliptic into transverse elliptic form, and the posterior part of pelvic inlet became larger.Sacral concavity was increased and the pelvis became deeper. Mid pelvis and the pelvic outlet was almost unchanged, and showed no correlation with age.

Placental homing peptide guides HIF1α­silenced exosomes conjugates for targeted enhancement of invasion of trophoblast cells.

There is a current lack of availability of therapeutics to treat Preeclampsia (PE), primarily due to the risk of harm to the fetus. Hypoxia­inducible factor­1α (HIF1α) is highly expressed in trophoblast cells and suppresses their invasive ability. Extensive studies have confirmed the positive effects of mesenchymal stem cell (MSC)­derived exosomes on PE. The aim of the present study was to develop a method for targeted delivery of HIF1α­silenced exosomes to the placenta. HIF1α was overexpressed in JEG­3 cells. Then, the glucose uptake, lactate production, proliferation and invasion of HIF1α­elevated JEG­3 cells were detected. Exosomal membrane protein lysosome­associated membrane glycoprotein 2b and placental homing peptide CCGKRK gene sequence amplified by PCR were conjugated using short hairpin RNA­HIF1α (sh­HIF1α) sequence (exo­pep­sh­HIF1α), which were then transfected into MSCs cultured in vitro. Exosomes were isolated from the supernatant of the aforementioned MSCs and identified by determining the size and exosomal markers. Finally, the invasion ability of MSCs­derived exosomes treated JEG­3 cells were detected using Transwell assays. HIF1α was demonstrated to remarkably promote the uptake of glucose and the production of lactate in JEG­3 cells. In addition, high levels of HIF1α facilitated the proliferation of JEG­3 cells, while suppressing their invasion ability. Bone marrow derived MSCs were cultured in vitro and exosomes were successfully isolated from these cells. Exo­pep­sh­HIF1α significantly reduced placental HIF1α expression, and induced significant enhancement of placental invasion. Overall, placental homing peptide­guided HIF1α­silenced exosomes effectively facilitated the invasion of placental trophoblasts, which could be used for the targeted delivery of payloads to the placenta and serve as a novel placenta­specific therapeutic approach.

The relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia.

OBJECTIVES: To investigate the relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia. MATERIAL AND METHODS: Real-time PCR (qRT-PCR) was used to detect the hsa_circ_0051326 expression level. Enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and western blotting were used to detect HLA-G expression in blood in 50 patients with pre-eclampsia and 50 normal pregnant women. RESULTS: HLA-G protein expression level was decreased significantly in the blood of patients with pre-eclampsia. hsa_circ_0051326 and HLA-G mRNA in blood were decreased significantly in the pre-eclampsia patients compared with normal pregnant women. There was a positive correlation between the expression of serum hsa_circ_0051326 with HLA-G mRNA. CONCLUSIONS: Serum hsa_circ_0051326 was a new diagnostic biomarker for pre-eclampsia with equivalent efficiency of HLA-G. Maternal serum hsa_circ_0051326 level may pre-diagnose potentially pre-eclampsia before delivery.