Identification and characterization of isocitrate dehydrogenase 1 (IDH1) as a functional target of marine natural product grincamycin B.

Grincamycins (GCNs) are a class of angucycline glycosides isolated from actinomycete Streptomyces strains that have potent antitumor activities, but their antitumor mechanisms remain unknown. In this study, we tried to identify the cellular target of grincamycin B (GCN B), one of most dominant and active secondary metabolites, using a combined strategy. We showed that GCN B-selective-induced apoptosis of human acute promyelocytic leukemia (APL) cell line NB4 through increase of ER stress and intracellular reactive oxygen species (ROS) accumulation. Using a strategy of combining phenotype, transcriptomics and protein microarray approaches, we identified that isocitrate dehydrogenase 1(IDH1) was the putative target of GCN B, and confirmed that GCNs were a subset of selective inhibitors targeting both wild-type and mutant IDH1 in vitro. It is well-known that IDH1 converts isocitrate to 2-oxoglutarate (2-OG), maintaining intracellular 2-OG homeostasis. IDH1 and its mutant as the target of GCN B were validated in NB4 cells and zebrafish model. Knockdown of IDH1 in NB4 cells caused the similar phenotype as GCN B treatment, and supplementation of N-acetylcysteine partially rescued the apoptosis caused by IDH1 interference in NB4 cells. In zebrafish model, GCN B effectively restored myeloid abnormality caused by overexpression of mutant IDH1(R132C). Taken together, we demonstrate that IDH1 is one of the antitumor targets of GCNs, suggesting wild-type IDH1 may be a potential target for hematological malignancies intervention in the future.

Tumor location of the central and nipple portion is associated with impaired survival for women with breast cancer.

BACKGROUND: Tumor location in the breast varies, with the highest frequency in the upper outer quadrant and lowest frequency in the lower inner quadrant. Nevertheless, tumors in the central and nipple portion (TCNP) are poorly studied types of breast cancer; therefore, we aimed to clarify the clinicopathological characteristics and prognostic features of TCNP. METHODS: Using the Surveillance, Epidemiology, and End Results database, we identifed 105,037 patients diagnosed with tumor in the breast peripheral quadrant (TBPQ) (n=97,046) or TCNP (n=7,991). The chi-squared test was used to compare categorical variables across TCNP and TBPQ. Cox proportional hazard models with hazard ratios were applied to estimate the factors associated with prognosis. RESULTS: The median follow-up was over 43 months. Compared with TBPQ, TCNP patients were signifcantly older (age ≥66 years: 40.4% vs 34.1%, P<0.001), with larger tumor sizes (>20 mm size: 46.9% vs 37.3%, P<0.001), higher proportions of TNM stage II-III (18.6% vs 9.9%, P<0.001), and more mastectomies (58.1% vs 37.8%, P<0.001). The breast cancer-specifc survival (BCSS)/overall survival (OS) rate was signifcantly worse for TCNP than for TBPQ. Multivariate Cox analysis showed a higher hazard ratios for TCNP over TBPQ (BCSS: hazard ratios =1.160, P=0.005, 95% CI: 1.046-1.287; OS: hazard ratios =1.301, P<0.001, 95% CI: 1.211-1.398). A subgroup analysis revealed inferior outcomes for TCNP in TNM stage II-III and breast subtype subgroup. Multivariate logistic regression indicated that TCNP was an independent contributing factor to LN metastasis. CONCLUSIONS: TCNP was associated with older age, larger tumor size, higher TNM stage, and lymph node metastasis. Compared with TBPQ, TCNP had adverse impacts on BCSS and OS.

Trace element profiles in pregnant women's sera and umbilical cord sera and influencing factors: Repeated measurements.

In utero exposure to toxic heavy metals and deficient or excessive essential trace elements during pregnancy may have adverse effects on pregnant women and their offsprings, which are of great concern. The objective of the present study was to characterize serum concentrations of multiple trace elements at multiple time points during pregnancy in Chinese women. Three thousand four hundred and sixteen pregnant women in total were included from MABC (Ma'anshan Birth Cohort) study. Fasting sera in the morning and questionnaires were obtained at three separate follow-up visits. Nineteen trace elements from serum samples were analyzed, including aluminum (Al), vanadium (V), chromium (Cr), manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), barium (Ba), thallium (Tl), lead (Pb), calcium (Ca), magnesium (Mg), mercury (Hg) and molybdenum (Mo). The total detection rates for most elements were 100% rather than Ni (99.98%), As (99.97%), Cd (99.6%), Ba (99.9%), Pb (99.8%), Hg (99.8%). The concentration distributions of 19 elements varied vastly. Median concentrations for all trace elements ranged from 38.5 ng/L to 102.9 mg/L. The moderate interclass correlation coefficients (ICCs) were observed for Co, Cu, Se and Hg, ranging from 0.40 to 0.62; the lower ICCs, ranging from 0.13 to 0.32 were for Fe, Zn, Cd, Ba, Tl, Mg and Mo. The intraclass correlation effects were not observed for the remaining elements, such as Al, V, Cr, Mn, Ni, As and Pb. The concentrations of each element between three time points were significantly different; significant differences were also found between any two time points except for Ni, Cd and Mo. Many factors could affect the levels of trace elements, and a very important factor of them was season. Consequently, a single measurement of elements in sera seems not enough to describe exposure levels throughout pregnancy; additionally, season affected exposure levels of trace elements with moderate ICCs showed certain regularity. Future analyses should take sampling seasons into consideration carefully.

Association between maternal and umbilical cord serum cobalt concentration during pregnancy and the risk of preterm birth: The Ma'anshan birth cohort (MABC) study.

Cobalt is an essential trace element and has been suggested to be involved in oxidative stress and inflammatory responses. However, researches have paid little attention to the association between serum cobalt levels during pregnancy and the risk of preterm birth (PTB, <37 week of gestation). The purpose of this study was to determine the association between maternal and umbilical cord serum cobalt concentrations and the risk of PTB. A total of 2951, 3080, and 2698 serum samples were obtained from pregnant women in the first, the second trimester, and the umbilical cord blood, respectively. The tertile levels of ln-transformed cobalt were defined as low, medium and high levels for cobalt respectively. In our study, the rate of PTB (<37 weeks of gestation) was elevated in subjects with low cobalt levels in the first trimester of pregnancy (adjusted OR 1.61, 95% CI: 1.01, 2.58) and the second trimester of pregnancy (adjusted OR 1.62, 95% CI: 1.03, 2.54). The adjusted OR for PTB was 2.46 (95% CI: 1.34, 4.53) among subjects with low cobalt levels and 2.22 (95% CI: 1.19, 4.15) among subjects with medium cobalt levels in the umbilical cord serum. Our findings demonstrated that the lower levels in maternal and umbilical cord serum cobalt were associated with the increased the risk of PTB.

Repeated measures of prenatal phthalate exposure and maternal hemoglobin concentration trends: The Ma'anshan birth cohort (MABC) study.

A prospective cohort study of a Chinese population was conducted to investigate the relationship between prenatal phthalates exposure and maternal hemoglobin or anemia. Based on the Ma'anshan Birth Cohort study, 7 phthalate metabolites were quantified in spot pregnancy urine samples (n = 9263) from 3269 pregnant women during each trimester. The maternal hemoglobin concentrations were obtained from electronic medical records at the same three time points for each participant during pregnancy. Anemia was defined as a hemoglobin concentration below 110 g/L in pregnant women. Repeated measures and trimester-specific analyses were used to estimate the effects of phthalates exposure on maternal hemoglobin and anemia. The prevalence of anemia was 3.6%, 27.0%, and 26.5% during the first, second and third trimester, respectively. Repeated measures analysis showed that hemoglobin concentrations decreased by 0.55, 0.19, 0.57, 0.49, and 0.54 g/L with each 1 ln-transformed concentration increase of MBP, MBzP, MEHP, MEOHP, and MEHHP, respectively. Exposure to MMP, MBP, MEHP, MEOHP, and MEHHP increased the risk of anemia by 1.11-fold, 1.21-fold, 1.20-fold, 1.13-fold, and 1.16-fold, respectively. Trimester-specific regression models stratified by the sample collection time during pregnancy generated consistent results. This is the first study focusing on the effect of prenatal phthalate exposures on hemoglobin or anemia in pregnant Chinese women. We found that prenatal phthalates exposure not only decreased the concentrations of hemoglobin but also showed associations with the prevalence of anemia. Associations appeared stronger for the subsample representing women pregnant with a male fetus than those pregnant with a female fetus. Anemia remains a moderate public health problem in China, and effective measures should be implemented.

Impact of maternal thyroid autoantibodies positivity on the risk of early term birth: Ma'anshan Birth Cohort Study.

PURPOSE: We aim to investigate associations of maternal serum anti-thyroperoxidase autoantibody (TPOAb) with duration of gestation. We aim to investigate whether maternal TPOAb positivity is associated with the risk of premature or early term birth. METHODS: This was a prospective birth cohort study performed in an iodine sufficient area of China. Serum samples were collected from 2931 women at both the first and second trimesters of pregnancy. Thyrotropin (TSH), free thyroxine (FT4), and TPOAb levels were measured. Data on gestational age at birth was obtained from delivery records. RESULTS: The prevalence of early term birth was 23.8%, while the prevalence of premature birth was 4.2%. The prevalence of TPOAb positivity was 12.1% in the first trimester and was 7.2% in the second trimester. Gestational age at birth was inversely associated with lgTPOAb both in the first trimester (ß, -0.283, 95% CI -0.408, -0.158; P < 0.001) and in the second trimester (ß, -0.174, 95% CI -0.319, -0.030; P = 0.018), after adjustment for potential confounding factors. There was a positive association of TPOAb positivity with the risk of early term birth both in the first (OR = 1.691, 95% CI 1.302, 2.197) and second trimesters (OR = 1.644, 95% CI 1.193, 2.264), after adjustment for potential confounding factors. TPOAb positivity in the second trimester was associated with a 1.863-fold higher risk of premature birth (OR = 1.863, 95% CI 1.009, 3.441), after adjustment for potential confounding factors. CONCLUSIONS: Our results show that TPOAb is associated with shorter duration of gestation and with higher risk of premature and early term birth.

Thyroid autoantibodies in pregnancy are associated with hypertensive disorders of pregnancy: Ma'anshan Birth Cohort Study.

OBJECTIVE: Hypertensive disorders of pregnancy (HDP) have been associated with adverse health outcomes for both mothers and children. Previous studies examining associations of maternal thyroid autoantibodies with HDP indicate conflicting results. The objective of this study was to examine associations of maternal thyroid autoantibody positivity in the first and the second trimesters with the risk of HDP. DESIGN, PARTICIPANTS AND MEASUREMENTS: In the Ma'anshan Birth Cohort study, a population-based prospective study in China, a total of 3474 pregnant women were enrolled between May 2013 and September 2014. Thyroid autoantibodies, including antithyroperoxidase autoantibody (TPOAb) and antithyroglobulin autoantibody (TgAb), as well as thyroid function tests, were measured in both the first and the second trimesters in 2893 pregnant women. Multivariate logistic regression analyses were conducted to calculate the odds ratio (OR) and 95% confidence interval (CI) for the associations between thyroid autoantibodies and HDP. RESULTS: Multivariate logistic regression analyses showed that TPOAb positivity in the first trimester was associated with a 1.80 (95% CI = 1.17-2.78) increased odds of HDP after adjustment for confounders, which was mainly due to an increased risk of gestational hypertension (OR = 1.93, 95% CI = 1.17-3.18). In addition, TgAb positivity in the first trimester was associated with a higher risk of HDP (OR = 1.78, 95% CI = 1.16-2.73) after adjustment for confounders, which was mainly due to an increased risk of gestational hypertension (OR = 1.89, 95% CI = 1.15-3.11). These associations were also seen among euthyroid women. Women with positive TPOAb in the second trimester seemed to have a higher risk of gestational hypertension (OR = 1.87, 95% CI = 1.02-3.43) after adjustment for confounders. However, among euthyroid women, TPOAb positivity in the second trimester was not associated with HDP. The TgAb status in the second trimester was not associated with HDP. CONCLUSIONS: Our results show that TPOAb positivity and TgAb positivity in the first trimester are associated with an increased risk of HDP. These data demonstrate that these associations are even seen among euthyroid women.

Prenatal phthalate exposure and placental size and shape at birth: A birth cohort study.

OBJECTIVE: There is concern over the potential placental effects of prenatal phthalate exposure, and the potential adverse effects of prenatal phthalate exposure require further study; however, few data are available in humans. We investigated the associations between phthalate exposure in each trimester and both placental size and shape at birth. METHODS: We measured the urinary concentrations of phthalate metabolites among 2725 pregnant women in the Ma'anshan Birth Cohort. Before collecting urine samples from each of the three trimesters, the pregnant women were interviewed via questionnaires. Placental information was obtained from hospital records. We estimated the sex-specific associations between urinary phthalate concentrations in each trimester and both placental size and shape at birth using adjusted multiple regression. A linear mixed model was used for the repeated measures analysis with subject-specific random intercepts and slopes for gestational age at sample collection to test the effect of phthalate levels on placental size and shape and to estimate the effect sizes. RESULTS: Overall, placental breadth increased by 0.148cm (95% CI: 0.078, 0.218) with each 1 ln-concentration increase in MBP in the first trimester. The difference between placental length and breadth (length-breadth) decreased by 0.086cm (95% CI: -0.159, -0.012) and 0.149cm (95% CI: -0.221, -0.076) with each 1 ln-concentration increase in MMP and MBP, respectively, in the first trimester. In the second trimester, placental thickness increased by 0.017cm (95% CI: 0.006, 0.027), 0.020cm (95% CI: 0.004, 0.036), 0.028cm (95% CI: 0.007, 0.048), and 0.035cm (95% CI: 0.018, 0.053) with each 1 ln-concentration increase in MMP, MBP, MEOHP, and MEHHP, respectively. In the third trimester, placental thickness increased by 0.037cm (95% CI: 0.019, 0.056) and 0.019cm (95% CI: 0, 0.037) with each 1 ln-concentration increase in MBP and MEHP, respectively. Multiple linear regression for each offspring sex indicated that prenatal phthalate exposure increased placental thickness in both the first and second trimesters in males, whereas the corresponding relationship was close to null in females. Linear mixed models (LMMs) yielded similar results. CONCLUSION: Our results suggest the presence of associations between prenatal phthalate exposure and placental size and shape. Exposure to certain phthalates may cause the placenta to become thicker and more circular. Associations appeared stronger for the subsample representing male offspring than those for the subsample representing female offspring. Given the few studies on this topic, additional research is warranted.

Incidence and risk factors of bacterial vaginosis among pregnant women: a prospective study in Maanshan city, Anhui Province, China.

AIM: The aim of this study was to identify the incidence and risk factors of bacterial vaginosis among pregnant women. MATERIAL AND METHODS: Prospective data from a cohort of 668 pregnant women were used to identify potential risk factors for bacterial vaginosis during pregnancy by Cox proportional hazards regression. RESULTS: A total of 204 incident cases of bacterial vaginosis were diagnosed in 274.8 woman-years of follow-up. The bacterial vaginosis incidence rate was 0.74 per 1 woman-year and median prevalence during follow-up was 15.6%. In the adjusted model, changing underwear nearly everyday, miscarriage history, urinary tract infection during follow-up, husbands' education level, and concurrent trichomoniasis and candidiasis remained significantly associated with bacterial vaginosis (adjusted hazard ratio and 95% confidence interval were 1.87 [1.26-2.77]; 2.96 [1.96-4.47]; 2.41 [1.05-5.49]; 0.50 [0.32-0.77]; 1.82 [1.02-3.25]; 1.88 [1.30-2.70], respectively). CONCLUSION: Bacterial vaginosis during pregnancy can be affected by many factors, and some are indirectly acting factors. Further prospective studies that include a larger sample size and more information on the development of bacterial vaginosis are needed.

Setdb2 controls convergence and extension movements during zebrafish gastrulation by transcriptional regulation of dvr1.

As the primary driving forces of gastrulation, convergence and extension (C&E) movements lead to a medio-lateral narrowing and an anterior-posterior elongation of the embryonic body axis. Histone methylation as a post-translational modification plays a critical role in early embryonic development, but its functions in C&E movements remain largely unknown. Here, we show that the setdb2-dvr1 transcriptional cascade plays a critical role in C&E movements during zebrafish gastrulation. Knockdown of Setdb2, a SET domain-containing protein possessing a potential histone H3K9 methyltransferase activity, induced abnormal C&E movements, resulting in anterior-posterior shortening and medio-lateral expansion of the embryonic axis, as well as abnormal notochord cell polarity. Furthermore, we found that Setdb2 functions through fine-tuning the expression of dvr1, a ligand of the TGF-ß superfamily, to an appropriate level to ensure proper C&E movements in a non-cell-autonomous manner. In addition, both overexpression and knockdown of Dvr1 at the one-cell stage resulted in defects at epiboly and C&E. These data demonstrate that Setdb2 is a novel regulator for C&E movements and acts by modulating the expression level of dvr1, suggesting that Dvr1 acts as a direct and essential mediator for C&E cell movements.

[Causation between occupational prolonged standing time during progestation and pregnancy related complications].

OBJECTIVE: To describe the epidemiological characteristics of profession related long-standing behavior during preconception and progestation, and to probe the relationship between prolonged standing jobs and the common pregnancy related complications among pregnant women, in Ma' anshan city,Anhui province. METHODS: In this cohort study, subjects who had their first antenatal examination at Ma' anshan Maternal and Child Care Centers were recruited under informed consent, from October 2008 to October 2010. All the information were collected through questionnaires in the first, second and third trimesters respectively. Three questionnaires were filled in by subjects under the guidance of healthcare takers. RESULTS: In the study, the 75(th) percentile of prolonged-standing times were 4.0 h/d and 3.0 h/d respectively in preconception and progestation. The characteristics of pregnant women with low social/economic status, prone to be involved in stand-long occupation. Results in logistic regression analysis, prolonged-standing jobs during preconception was the risk factor of pregnancy-induced hypertension and severe anemia. The adjusted odds ratios were 2.05 (95%CI:1.26-3.31) and 1.38(95%CI:1.03-1.85)respectively. CONCLUSION: Prolonged standing jobs appeared to be common occupational exposure to and risks of both pregnant woman and their fetus. Exposure to these kinds of jobs during preconception could increase the risk of pregnancy-induced hypertension and severe anemia. In order to promote maternal health programs, all the related occupational risk factors should be valued and avoided during preconception and pregnancy.

[Relationship between folic acid supplements during peri-conceptional period and the adverse pregnancy outcomes: a cohort study].

OBJECTIVE: To examine the association between folic acid supplements during peri-conception and the related adverse birth outcome. METHODS: Pregnant women who received first prenatal care at 4 municipal-level medical institutions in Maanshan, from Oct. 2008 to Oct. 2010 were selected as the target population. All participants were asked to complete a self-administered questionnaire which including data on demographic characteristics, interval, complications and frequency of taking folic acid etc., during pregnancy. The follow-up-records after delivery would include factors as: fetal weight, height, circumference of head, chest circumference of the neonates. Finally, 4448 valid questionnaires were gathered, including 190 premature, 147 small for gestational age and 104 low birth weight babies. Descriptive statistics and logistic regression models were used for data analysis. RESULTS: Data showed that the weight, height and head circumference of the fetal at birth among pregnant women who had taken supplementary standard folic acid during peri-conception period or only during the first trimester, were all better than those pregnant women who had not taken the standard folic acid supplements. After adjustment for potential confounders as gestational weeks, maternal age, mather's education level, results from the logistic regression showed that intake of standard folic acid supplements appeared a protective factor for those babies who were smaller than the gestational age (RR = 0.45, 95%CI: 0.24 - 0.86), at premature delivery (RR = 0.52, 95%CI: 0.32 - 0.87) or with low birth weight (RR = 0.39, 95%CI: 0.19 - 0.80). However, data from this study showed that provision of folic acid supplements to the pre-pregnant or at first trimester alone did not make obvious impact on those babies as prematured, small for gestational age and at low birth weight. CONCLUSION: Standardized provision of folic acid supplements during peri-conceptional period could improve the outcomes of birth.

Activated N-Ras signaling regulates arterial-venous specification in zebrafish.

BACKGROUND: The aberrant activation of Ras signaling is associated with human diseases including hematological malignancies and vascular disorders. So far the pathological roles of activated Ras signaling in hematopoiesis and vasculogenesis are largely unknown. METHODS: A conditional Cre/loxP transgenic strategy was used to mediate the specific expression of a constitutively active form of human N-Ras in zebrafish endothelial and hematopoietic cells driven by the zebrafish lmo2 promoter. The expression of hematopoietic and endothelial marker genes was analyzed both via whole mount in situ hybridization (WISH) assay and real-time quantitative PCR (qPCR). The embryonic vascular morphogenesis was characterized both by living imaging and immunofluorescence on the sections with a confocal microscopy, and the number of endothelial cells in the embryos was quantified by flow cytometry. The functional analyses of the blood circulation were carried out by fluorescence microangiography assay and morpholino injection. RESULTS: In the activated N-Ras transgenic embryos, the primitive hematopoiesis appeared normal, however, the definitive hematopoiesis of these embryos was completely absent. Further analysis of endothelial cell markers confirmed that transcription of arterial marker ephrinB2 was significantly decreased and expression of venous marker flt4 excessively increased, indicating the activated N-Ras signaling promotes the venous development at the expense of arteriogenesis during zebrafish embryogenesis. The activated N-Ras-expressing embryos showed atrophic axial arteries and expansive axial veins, leading to no definitive hematopoietic stem cell formation, the blood circulation failure and subsequently embryonic lethality. CONCLUSIONS: Our studies revealed for the first time that activated N-Ras signaling during the endothelial differentiation in vertebrates can disrupt the balance of arterial-venous specification, thus providing new insights into the pathogenesis of the congenital human vascular disease and tumorigenic angiogenesis.

Identification of zinc-finger BED domain-containing 3 (Zbed3) as a novel Axin-interacting protein that activates Wnt/beta-catenin signaling.

Axin, a key modulator of the Wnt/beta-catenin pathway, acts as a scaffold protein in phosphorylating and degrading cytoplasmic beta-catenin. Canonical Wnt proteins appear to stabilize beta-catenin by inducing the interaction of LRP5/6 with Axin. This interaction requires the phosphorylation of the Ser or Thr residues in the PPPP(S/T)PX(T/S) motifs at the intracellular domain of LRP5/6. In this work, we identified a novel Axin-interacting protein, zinc-finger BED domain-containing 3 (Zbed3), by yeast two-hybrid screening. The interaction was confirmed in co-immunoprecipitation experiment in mammalian cells and in vitro pulldown assays. Moreover, we found Zbed3 also contains a PPPPSPT motif, which is crucial to its binding to Axin. The Ser and Thr residues in the motif appear to be also phosphorylated by glycogen synthase kinase 3beta (GSK3beta) and the CKI family kinases, as GSK3beta and CKIepsilon could enhance the interaction of Zbed3 with Axin. Mutation of the Ser (SA) or Thr (TA) residue to Ala in the motif markedly impaired its ability to interact with Axin. Expressing Zbed3, but not these mutants, led to inhibition of GSK3beta-mediated beta-catenin phosphorylation, cytoplasmic beta-catenin accumulation, and activation of lymphoid enhancer binding factor-1-dependent reporter gene transcription. Furthermore, knockdown of Zbed3 with RNA interference attenuated Wnt-induced beta-catenin accumulation, lymphoid enhancer binding factor-1-dependent luciferase reporter activity, and the Wnt target gene expression. These results together indicate that Zbed3 is a novel Axin-binding protein that is involved in Wnt/beta-catenin signaling modulation.

Isoflavones regulate secretion of leukemia inhibitory factor and transforming growth factor {beta} and expression of glycodelin in human endometrial epithelial cells.

Isoflavones have attracted much attention due to their association with health benefits; however, comprehensive understanding of the beneficial impacts of isoflavones on uterine biology at the molecular level remains unexplored. In the present study, our data showed that isoflavones aglycones AglyMax, genistein, and equol, but not daidzein, within the range of plasma concentration, displayed bioavailability in regulating the secretion of leukemia inhibitory factor (LIF) and transforming growth factor beta (TGF-beta) in Ishikawa cells, which was blocked by an estrogen receptor antagonist ICI 182 780, mitogen-activated protein kinase kinase (MEK)1/2 inhibitor PD98059, and p38 mitogen-activated protein kinase inhibitor SB203580. We also found that AglyMax and genistein increased in cyclic AMP release and the expression of glycodelin protein in Ishikawa cells assayed using western blot and immunochemical staining. The MEK1/2 inhibitor PD98059 and the protein kinase A inhibitor H89, but not SB203580, attenuated this glycoprotein expression. Moreover, isoflavone aglycones AglyMax stimulated LIF, and TGF-beta secretion, and glycodelin expression in separate primary endometrial epithelial cells in the follicular phase or luteal phase from healthy subject donors. Overall, our findings suggest that isoflavones may alter the uterine expression of estrogen-responsive genes.

Characterization of function of three domains in dishevelled-1: DEP domain is responsible for membrane translocation of dishevelled-1.

Wnt signaling plays an important role in embryogenesis and tumorgenesis. Although the mechanism about how Wnts transduce their signaling from receptor frizzled (Fz) to cytosol has not been understood, dishevelled (Dvl) protein was considered as the intersection of Wnt signal traffic. In this study, we characterized the function of three domains (DIX, PDZ and DEP) of Dvl-1 in canonical Wnt signal transduction and Dvl-1 membrane translocation. It was found both DIX and DEP domain were sufficient to block Wnt-3a-induced LEF-1 transcriptional activity and free cytosol beta-catenin accumulation; whereas PDZ domain and a functional mutant form of DEP domain (DEP-KM) had no effect on canonical Wnt signaling. In addition, when cotransfected with Fz-7, DEP domain, but not DIX, PDZ or DEP-KM, translocated and co-localized with Fz-7 to the plasma membrane, which was similar to Dvl-1. Furthermore, it was DEP domain that could block Fz-7-induced membrane translocation of Dvl-1 via a possible competitive mechanism. These results strongly suggest that DEP domain is responsible for the membrane translocation of Dvl-1 protein upon Wnt signal stimulation.