HDAC2-mediated proliferation of trophoblast cells requires the miR-183/FOXA1/IL-8 signaling pathway.

Pre-eclampsia (PE) is a major cause of maternal and perinatal death. Previous research has indicated the role of histone deacetylase 2 (HDAC2) in the pathogenesis of PE but the relevant molecular mechanisms are unknown. However, there is hitherto little information concerning the molecular mechanism behind HDAC2 in PE. Herein, we hypothesized that HDAC2 promotes trophoblast cell proliferation and this requires the involvement of microRNA-183 (miR-183), forkhead box protein A1 (FOXA1), and interleukin 8 (IL-8). We collected placental specimens from 30 PE affected and 30 normal pregnant women. HDAC2 and FOXA1 were poorly expressed while miR-183 and IL-8 were highly expressed in placental tissues in PE. In vitro, HDAC2 overexpression enhanced the proliferation, migration, and invasion of human trophoblast cells HTR-8/SVNEO. HDAC2 inhibited the expression of miR-183 by diminishing H4 acetylation in the miR-183 promoter region. miR-183 inhibition by its specific inhibitor increased the expression of FOXA1 and thus enhanced HTR-8/SVNEO cell proliferation, migration, and invasion. FOXA1, a transcriptional factor, enhanced HTR-8/SVNEO cell proliferation, migration, and invasion by inhibiting the transcription of IL-8. We also observed HDAC2 knockdown was lost upon FOXA1 overexpression, suggesting that HDAC2 could promote HTR-8/SVNEO proliferation, migration, and invasion through the miR-183/FOXA1/IL-8 pathway. In summary, the results highlighted the role of the HDAC2/miR-183/FOXA1/IL-8 pathway in PE pathogenesis and thus suggest a novel molecular target for PE.

DDP-resistant ovarian cancer cells-derived exosomal microRNA-30a-5p reduces the resistance of ovarian cancer cells to DDP.

Exosomes carrying microRNAs (miRNAs) have been demonstrated to play critical roles in the regulation of development, growth and metastasis of cancer. Bioinformatic predictions identified differentially expressed SRY-box 9 (SOX9) in OC, and the regulatory miRNA miR-139-5p. Here, we aim to evaluate the function of exosomal miR-139-5p in the sensitivity of ovarian cancer (OC) cells to cis-diamminedichloroplatinum(II) (DDP). Expression pattern of miR-139-5p and SOX9 in ovarian cancer cells (SKOV3) and DDP-resistant cells (SKOV3/DDP) was identified using reverse transcription quantitative polymerase chain reaction and western blot analysis. The relationship between miR-139-5p and SOX9 was validated using a dual-luciferase reporter assay. SKOV3/DDP cell line was developed and introduced with miR-30a-5p mimic to analyse the effects of miR-30a-5p on resistance to DDP. The in vitro and in vivo effects of exosomal miR-30a-5p on resistance of SKOV3 cells to DDP were assessed in a co-culture system of exosomes and OC cells as well as in tumour-bearing nude mice. High expression of SOX9 and low expression of miR-30-5p were witnessed in OC. Furthermore, miR-30-5p, a downregulated miRNA in SKOV3/DDP cells, increased the rate of cell apoptosis and enhanced the sensitivity of SKOV3/DDP cells to DDP by targeting SOX9. Moreover, exosomes carrying miR-30a-5p were identified to sensitize SKOV3/DDP cells to DDP both in vitro and in vivo. These data together supported an important conclusion that DDP-resistant OC cell-derived exosomal miR-30a-5p enhanced cellular sensitivity to DDP, highlighting a potential strategy to overcome drug resistance.

Sargassum integerrimum inhibits oestrogen deficiency and hyperlipidaemia-induced bone loss by upregulating nuclear factor (erythroid-derived 2)-like 2 in female rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Oestrogen deficiency, high incidences of hyperlipidaemia (HLP) and accelerated bone loss frequently occur in postmenopausal women. There is an urgent need to develop functional foods or specific drugs to protect against bone loss induced by oestrogen deficiency with HLP. AIM OF THE STUDY: In this study, we investigated the potential inhibitory effects of Sargassum integerrimum (SI) on bone loss in an ovariectomized rat model with HLP. MATERIALS AND METHODS: The rats were treated for 12 weeks, and then, bone mineral density, bone biomechanical, bone microstructure, bone morphology, biomarkers of HLP oxidative stress and side effects were determined. Immunohistochemical staining and Western blot were performed to evaluate related protein expression. RESULTS: The femur bone mineral density increased (P < 0.05), and the microscopic structures (ratio of bone volume to total volume [BV/TV], connectivity density [Conn.D], trabecular number [Tb.N] and trabecular thickness [Tb.Th]) of the bone trabecula and mechanical properties (maximum and breaking load [ML and BL, respectively]) improved after SI treatment (P < 0.05). Furthermore, the levels of HLP biomarkers (total cholesterol, triglyceride and low-density lipoprotein) were significantly decreased (P < 0.05), whereas the levels of antioxidant markers (superoxide dismutase and total antioxidant capacity) were increased (P < 0.05). Similar results were obtained with immunohistochemical staining, whereas the Western blot assay showed that SI stimulated the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) in bone. CONCLUSION: Our data indicate that rats exposed to SI treatment for 12 weeks did not exhibit noticeable side effects. In conclusion, SI suppressed bone loss induced by ovariectomized and the associated HLP in rats by activating Nrf2, which could be a promising treatment option for osteoporosis induced by oestrogen deficiency and HLP in postmenopausal women. TRANSLATIONAL SCOPE STATEMENT: Our study verified that SI prevented bone loss in rats with oestrogen deficiency with HLP by upregulating nuclear factor (erythroid-derived 2)-like 2. Furthermore, no side effect was observed after the long-term administration of SI. Those results suggested SI could be developed as a functional food or drug for postmenopausal osteoporosis induced by oestrogen deficiency with HLP.

Long Noncoding RNA LINC01133 Confers Tumor-Suppressive Functions in Ovarian Cancer by Regulating Leucine-Rich Repeat Kinase 2 as an miR-205 Sponge.

Ovarian cancer represents one of the most commonly occurring malignant tumors among females. Long noncoding RNAs act as biomarkers associated with the pathophysiology of multiple kinds of malignancies, including ovarian cancer. This study aimed to clarify the molecular mechanism of LINC01133 in the progression of ovarian cancer. Initially, microarray-based analysis was used to screen differentially expressed long noncoding RNAs and miRNAs, with LINC01133 and miR-205 obtained for this study. The biological functions of LINC01133, miR-205, and leucine-rich repeat kinase 2 (LRRK2) were validated on cell proliferation, migration, and invasion of ovarian cancer through gain-of-function and loss-of-function experiments. Finally, tumorigenesis was measured in vivo by inducing tumor xenograft in nude mice. The findings revealed a poor expression of LINC01133 in ovarian cancer tissue and cell, which was predominantly expressed in the cytoplasm. LINC01133 repressed cell proliferation, invasion, migration, and tumorigenic ability. miR-205 targeted and negatively regulated LRRK2. LINC01133 was also found to function as an miR-205 sponge to decrease ovarian cancer cell proliferation, migration, and invasion by elevating LRRK2. These in vitro findings were reproduced in vivo on tumor xenograft in nude mice. In conclusion, because of its role as an miR-205 sponge, LINC01133 repressed the development of ovarian cancer by up-regulating LRRK2.

Clinical significance and biological role of Wnt10a in ovarian cancer.

Ovarian cancer is one of the five most malignant types of cancer in females, and the only currently effective therapy is surgical resection combined with chemotherapy. Wnt family member 10A (Wnt10a) has previously been identified to serve an oncogenic function in several tumor types, and was revealed to have clinical significance in renal cell carcinoma; however, there is still only limited information regarding the function of Wnt10a in the carcinogenesis of ovarian cancer. The present study identified increased expression levels of Wnt10a in two cell lines, SKOV3 and A2780, using reverse transcription-polymerase chain reaction. Functional analysis indicated that the viability rate and migratory ability of SKOV3 cells was significantly inhibited following Wnt10a knockdown using short interfering RNA (siRNA) technology. The viability rate of SKOV3 cells decreased by ~60% compared with the control and the migratory ability was only ~30% of that in the control. Furthermore, the expression levels of ß-catenin, transcription factor 4, lymphoid enhancer binding factor 1 and cyclin D1 were significantly downregulated in SKOV3 cells treated with Wnt10a-siRNA3 or LGK-974, a specific inhibitor of the canonical Wnt signaling pathway. However, there were no synergistic effects observed between Wnt10a siRNA3 and LGK-974, which indicated that Wnt10a activated the Wnt/ß-catenin signaling pathway in SKOV3 cells. In addition, using quantitative PCR, Wnt10a was overexpressed in the tumor tissue samples obtained from 86 patients with ovarian cancer when compared with matching paratumoral tissues. Clinicopathological association analysis revealed that Wnt10a was significantly associated with high-grade (grade III, P=0.031) and late-stage (T4, P=0.008) ovarian cancer. Furthermore, the estimated 5-year survival rate was 18.4% for patients with low Wnt10a expression levels (n=38), whereas for patients with high Wnt10a expression (n=48) the rate was 6.3%. The results of the present study suggested that Wnt10a serves an oncogenic role during the carcinogenesis and progression of ovarian cancer via the Wnt/ß-catenin signaling pathway.

Response surface methodology for ultrasound-assisted extraction of astaxanthin from Haematococcus pluvialis.

Astaxanthin is a novel carotenoid nutraceutical occurring in many crustaceans and red yeasts. It has exhibited various biological activities including prevention or amelioration of cardiovascular disease, gastric ulcer, hypertension, and diabetic nephropathy. In this study, ultrasound-assisted extraction was developed for the effective extraction of astaxanthin from Haematococcus pluvialis. Some parameters such as extraction solvent, liquid-to-solid ratio, extraction temperature, and extraction time were optimized by single-factor experiment and response surface methodology. The optimal extraction conditions were 48.0% ethanol in ethyl acetate, the liquid-to-solid ratio was 20:1 (mL/g), and extraction for 16.0 min at 41.1 °C under ultrasound irradiation of 200 W. Under optimal conditions, the yield of astaxanthin was 27.58 ± 0.40 mg/g. The results obtained are beneficial for the full utilization of Haematococcus pluvialis, which also indicated that ultrasound-assisted extraction is a very useful method for extracting astaxanthin from marine life.

[Relationship between total bile acid concentration and fetal pulmonary surfactant in intrahepatic cholestasis of pregnancy].

OBJECTIVE: To explore the relationship between total bile acid (TBA) concentration and fetal pulmonary surfactant in intrahepatic cholestasis of pregnancy (ICP). METHODS: Fifty five patients with ICP (ICP group) who received cesarean section from April 2008 to February 2010 in Second Xiangya Hospital, Central South University, were recruited. The general conditions of the neonates within 7 days after birth in ICP group were recorded. Those with fetal distress, neonatal asphyxia, or neonatal respiratory distress syndrome were referred as pathological neonates, others were referred as normal neonates. Over the same period, 23 healthy gravidas were recruited as control group. Enzymatic method was used to detect the TBA concentrations in maternal blood, cord blood and amniotic fluid. ELISA was employed to measure the urfactant protein A (SP-A) concentration in cord blood. High performance liquid chromatography system was used to detect the concentrations of phosphatidylcholine (PC), phosphatidylinositol (PI), lysophosphatidylcholine (LPC), and sphingomyelin (SM) in amniotic fluid. RESULTS: (1) The concentrations of TBA in maternal blood, cord blood and amniotic fluid were (30.1 ± 7.9), (9.3 ± 3.3) and (4.4 ± 1.5) mmol/L in ICP group, (4.8 ± 2.2), (4.9 ± 0.9) and (1.4 ± 1.1) mmol/L in control group, respectively. The differences between the two groups were significant (P < 0.05). (2) The SP-A concentration in cord blood in ICP group was (29.5 ± 6.4) µg/L, significantly higher than that in control group, which was (22.6 ± 7.4) µg/L (P < 0.05). (3) There were 20 pathological neonates and 35 normal neonates in ICP group. In pathological neonates, the concentrations of TBA and SP-A in cord blood were (10.9 ± 2.2) mmol/L, (37.0 ± 5.9) µg/L, respectively; and were (8.0 ± 2.8) mmol/L, (26.7 ± 4.8) µg/L in normal neonates. The differences were significant (P < 0.05). (4) There was a positive correlation between TBA concentration in cord blood and in maternal blood (r(1) = 0.706, P < 0.05). The TBA concentration in cord blood was positively correlated with SP-A concentration as well (r(3) = 0.494, P < 0.05). (5) The PC and PI concentrations in amniotic fluid were (65.4 ± 7.2) mg/L and (3.8 ± 0.6) mg/L in ICP group, (69.7 ± 3.7) mg/L and (4.3 ± 0.7) mg/L in control group, respectively. The differences were significant (P < 0.05). The concentration of LPC in amniotic fluid in ICP group was (4.8 ± 0.9) mg/L, significantly higher than that in control group (P < 0.05), which was (4.2 ± 0.6) mg/L. The concentration of SM in amniotic fluid was (3.5 ± 0.8) mg/L in ICP group, (4.0 ± 0.5) mg/L in control group, with no significant difference (P > 0.05). (6)The ratio of PC/LPC in ICP group (14.2 ± 3.2) was significantly lower than that in control group (16.9 ± 2.5) (P < 0.05). (7)The TBA concentration in cord blood was negatively correlated with PC and PI concentrations (r(1) = -0.561, r(2) = -0.407, P < 0.05), and had no correlation with LPC concentration (r(3) = 0.260, P > 0.05). CONCLUSIONS: (1) The fetal TBA concentrations in both cord blood and amniotic fluid of patients with ICP was higher than those of healthy gravidas, they were also positively correlated with maternal TBA concentration. (2) ICP resulted in the change of fetal pulmonary surfactant and this change was associated with TBA concentrations in both cord blood and amniotic fluid.