A prognostic index model for predicting long-term recurrence of uterine leiomyoma after myomectomy.

INTRODUCTION: Uterine leiomyoma (UL) is a common benign pelvic tumor in women that has a high recurrence rate. Our aim is to propose a prognostic index (PI) model for predicting the long-term recurrence risk of uterine leiomyoma (UL). METHODS: A total of 725 women who underwent myomectomy were enrolled in this retrospective multicenter study. Patients were contacted for follow-up. A PI model was proposed based on the multivariate Cox regression analysis in the model group. The predictive value of this model was tested in both internal and external validation group. RESULTS: PI formula = 1.5(if 3-5 leiomyomas) or 2(if >5 leiomyomas)+1(if residue)+1(if not submucosal)+1(if combined endometriosis). The PI value was divided into low-risk, intermediate-risk, and high-risk group by cut-off values 1.25 and 3.75. In the model group, the high-risk group had a significantly 4.55 times greater recurrence risk of UL than that in the low-risk group [cumulative recurrence rate (CR): 82.1% vs 29.5%, HR = 4.55, 95% CI 2.821-7.339]; the intermediate-risk group had a significantly 2.81 times greater recurrence risk of UL than that in the low-risk group (CR: 62.3% vs 29.5%, HR = 2.81, 95% CI 2.035-3.878). The differences between any two risk groups were also significant (P< 0.05) in both internal and external validation groups. CONCLUSION: The model was proved to be effective in predicting recurrence of UL after myomectomy.

Dienogest as a Maintenance Treatment for Endometriosis Following Surgery: A Systematic Review and Meta-Analysis.

This study aimed to comprehensively assess the value of Dienogest (DNG) as a maintenance treatment following conservative surgery for endometriosis in terms of the outcomes of disease and pregnancy. We searched for relevant studies and trials up to November 2020 from PubMed, Cochrane Library, Medline, and EMBASE databases as well as the Web of Science. Patients who received DNG maintenance treatment were compared to those who received other treatments (OT), including the levonorgestrel-releasing intrauterine system (LNG-IUS) and gonadotropin-releasing hormone analogs (GnRH-a), or non-treatment (NT). The primary outcomes were disease recurrence and pregnancy rates. Eleven studies were included in this meta-analysis. The pooled analysis indicated that DNG maintenance treatment was associated with a lower rate of disease recurrence. A significant difference was observed in DNG maintenance treatment compared with NT, but not with OT, in the pregnancy rates postoperatively. Moreover, DNG maintenance treatment was related to a significant increase in vaginal bleeding and weight gain. DNG can be recommended as a maintenance treatment for patients with endometriosis to decrease the rates of disease recurrence following conservative surgery. However, DNG maintenance treatment has no advantage in improving pregnancy rates compared to OT.

Staging procedures fail to benefit women with borderline ovarian tumours who want to preserve fertility: a retrospective analysis of 448 cases.

BACKGROUND: To evaluate the effect of clinicopathologic factors on the prognosis and fertility outcomes of BOT patients. METHODS: We performed a retrospective analysis of BOT patients who underwent surgical procedures in West China Second University Hospital from 2008 to 2015. The DFS outcomes, potential prognostic factors and fertility outcomes were evaluated. RESULTS: Four hundred forty-eight patients were included; 52 recurrences were observed. Ninety-two patients undergoing FSS achieved pregnancy. No significant differences in fertility outcomes were found between the staging and unstaged surgery groups. Staging surgery was not an independent prognostic factor for DFS. Laparoscopy resulted in better prognosis than laparotomy in patients with stage I tumours and a desire for fertility preservation. CONCLUSION: Patients with BOT fail to benefit from surgical staging. Laparoscopy is recommended for patients with stage I disease who desire to preserve fertility. Physicians should pay more attention to risk of recurrence in patients who want to preserve fertility.

Para-aortic lymphadenectomy did not improve overall survival among women with type I endometrial cancer.

OBJECTIVE: To compare outcomes and prognosis among women with type I endometrial cancer undergoing hysterectomy and bilateral salpingo-oophorectomy (H-BSO) with or without systematic pelvic lymphadenectomy (PLD) or para-aortic lymphadenectomy (PALD). METHODS: Retrospective review of women postoperatively diagnosed with type I endometrial cancer who underwent H-BSO at a university hospital in Chengdu, China (January 2010 to June 2012). Women were divided into no lymphadenectomy (PLD-/PALD-), systematic pelvic lymphadenectomy (PLD+/PALD-), or combined pelvic and para-aortic lymphadenectomy (PLD+/PALD+) groups. Follow-up was by telephone. Postoperative outcomes and prognosis were compared and risk factors were analyzed. RESULTS: In total, 333 women met the inclusion criteria: 121 underwent PLD+/PALD-, 166 underwent PLD+/PALD+, and 46 underwent PLD-/PALD-. There were no differences in pre-operative characteristics among the groups (all P>0.05). The PLD+/PALD+ group had a higher laparotomy rate (P=0.001), the PLD-/PALD- group had shorter operation time (P=0.001) and lower blood loss (P<0.001). There were no differences between the PLD+/PALD- and PLD+/PALD+ groups. Overall, 291 women had sufficient follow-up data; there was no difference in overall survival, and PALD was not a predictor of survival. CONCLUSION: Postoperative outcomes were similar among all surgical groups; a survival benefit of PALD was not demonstrated.

HOXB4 promotes the malignant progression of ovarian cancer via DHDDS.

BACKGROUND: Homeobox B4 (HOXB4) is correlated with poor prognosis of various cancer types. However, how HOXB4 promotes ovarian cancer (OV) progression remains unclear. METHODS: The Cancer Genome Atlas (TCGA) database indicated that a high level of HOXB4 in OV was correlated with poor prognosis. The biological functions of HOXB4 were confirmed by colony formation, migration, and invasion assays. The effect of HOXB4 on the expression of EMT cell markers was determined. The transcriptional target of HOXB4 was DHDDS, which was detected by a ChIP assay. A xenograft tumor model was generated in nude mice to detect the role of HOXB4 in tumor proliferation and metastasis. RESULTS: The results showed that HOXB4 protein levels were higher in OV tissues than in normal tissues and correlated with poor prognosis of OV. HOXB4 reduction inhibited the proliferation and invasion ability of OV cells in vitro. Conversely, these effects were enhanced by the upregulation of HOXB4 in OV cells. The binding of HOXB4 to two DNA motifs regulated DHDDS expression and contributed to the malignant progression of OV. The role of HOXB4 in contributing to tumor development in vivo was verified in mice. Further results indicated that HOXB4 induced Snail and Zeb1 expression. CONCLUSION: Overall, HOXB4 overexpression was remarkably correlated with poor prognosis of OV. Mechanistically, HOXB4 enhances the proliferation and invasion of tumor cells by activating DHDDS, thereby promoting the malignant progression of OV.

Role of epithelial-mesenchymal transition regulated by twist basic helix-loop-helix transcription factor 2 (Twist2) in embryo implantation in mice.

In a previous study we found the expression of epithelial-mesenchymal transition (EMT) biomarkers, including E-cadherin and N-cadherin, was significantly altered in uterine endometrium during embryo implantation via regulation by microRNA (miRNA)-429 and protocadherin-8 (Pcdh8). As a natural continuation of the previous study, the aim of the present study was to explore the role of EMT during embryo implantation and the potential activity of twist basic helix-loop-helix transcription factor 2 (Twist2) in regulating embryo implantation. A pregnancy model was established by naturally mating adult female ICR mice with fertile males. A pseudopregnancy model was established by mating fertile female ICR mice with vasectomised males. An invitro model of embryo implantation was established by the coculture of Ishikawa and JAR spheroids. Endometrial tissue during the peri-implantation period was collected, as were Ishikawa cells, JAR cells and cocultured cells. The expression of EMT markers (E-cadherin, N-cadherin, vimentin and cytokeratin) and Twist2 was detected invivo and invitro using the western blot analysis during embryo implantation. The expression of N-cadherin and vimentin (mesenchymal markers) was upregulated in the invitro implantation model, with downregulation of E-cadherin and cytokeratin (epithelial markers) expression. The expression of N-cadherin, vimentin and Twist2 increased significantly at the implantation sites at the time of implantation (Day 5), whereas the expression of E-cadherin and cytokeratin decreased. Location of Twist2 during embryo implantation was detected by immunohistochemistry (IHC), which revealed that it was extensively expressed in endometrial glandular epithelium and luminal epithelium at implantation sites on Day 5. The effect of the expression of Twist2 on embryo implantation was evaluated by suppressing Twist2 using Twist2-short interference (si) RNA in invivo and invitro models. The numbers of implanted embryos and the implantation rate were compared invivo and invitro. Western blot analysis showed that suppression of Twist2 led to upregulation of E-cadherin and cytokeratin, accompanied by downregulation of N-cadherin and vimentin (P<0.05). The number of implanted embryos after Twist2-siRNA interference was lower than in normal pregnancy (mean (±s.d.) 2.4±0.5 vs 6.8±1.3 respectively; P<0.05). These findings suggest the involvement of EMT in embryo implantation. The suppression of Twist2 could suppress embryo implantation by regulating EMT.

MicroRNA-30a-3p regulates epithelial-mesenchymal transition to affect embryo implantation by targeting Snai2†.

OBJECTIVE: To study the potential role of miR-30a-3p in embryo implantation and explore underlying mechanisms. METHODS: We first established normal pregnancy, pseudopregnancy, delayed implantation, and artificial decidualization mouse models. Next, we detected miR-30a-3p expression profiles of these models with real-time reverse transcription PCR(qRT-PCR), then predicted potential target genes through a dual-luciferase assay. Immunofluorescence-fluorescence in situ hybridization co-located miR-30a-3p and target genes. We then examined the effect of miR-30a-3p on embryo implantation in vivo and in vitro. Wound healing and transwell assays were employed to explore possible miR-30a-3p effects on epithelial-mesenchymal transition (EMT), before molecules related to the latter process were examined with qRT-PCR. RESULTS: MiR-30a-3p expression decreased significantly on embryo implantation day, compared with the peri-implantation period (P < 0.05). Identified target gene Snai2 expression increased significantly during implantation (P < 0.05). In vivo and in vitro analysis showed that up-regulation of miR-30a-3p by agomir and mimics resulted in decreased implantation sites and embryo implantation rate. Transfection of miR-30a-3p mimics to HEC-1-b cells decreased expression of Snai2 and mesenchymal markers (Vimentin and N-cadherin). Furthermore, wound healing area decreased, as did migration and invasion capacity. CONCLUSION: MiR-30a-3p is down-regulated in the embryo implantation period and might have some effect on embryo implantation by acting as a suppressor of EMT through targeting Snai2.

Expression of tissue factor in human cervical carcinoma tissue.

The present study aimed to investigate tissue factor (TF) expression in cervical cancer and explore its association with disease progression. A total of 258 cervical cancer tissues and their adjacent normal tissues were collected between September 2014 and September 2016. TF expression was detected in the tissue samples by immunohistochemistry and western blot analysis. Associations between the expression of TF and clinical stage, differentiation status and metastasis of cancer cells were examined. The mean immunohistochemistry score of TF expression in cervical cancer tissues was 2.86±1.76, which was significantly increased compared with the adjacent normal tissues (0.28±0.45). The expression of TF was also significantly associated with the clinical stage, lymph node metastasis and distant metastasis of cancer cells. Immunohistochemistry staining and western blot analysis demonstrated that TF expression in cervical cancer tissues significantly increased as the clinical stage increased. TF expression in tumor tissues from patients with lymph node metastasis was significantly increased compared with samples from patients without lymph node metastasis. TF expression was also significantly increased in patients with distant metastasis compared with those without. In conclusion, TF is highly expressed in cervical cancer tissues and high expression of TF may enhance the invasion and metastasis of cervical cancer cells.

Single-incision laparoscopic hysterectomy using conventional laparoscopic instruments: initial experience with 25 cases.

PURPOSE: To evaluate the feasibility and safety of single-incision laparoscopic hysterectomy using conventional instruments. METHODS: Twenty-five patients undergoing single-incision laparoscopic hysterectomy (SI-LAH) using conventional instruments at West China Second University Hospital between July, 2017 and December, 2017 were selected for participation. Another 25 cases undergoing traditional multi-port laparoscopic hysterectomy (MP-LAH) matched with similar uterine size were selected as controls. Characteristics and clinical data of patients including operative time, estimated blood loss, hospital stay, catheter retention time, and intraoperative and postoperative complications were compared between the two groups. RESULTS: The baseline characteristics of the two groups were comparable. The estimated blood loss was less in SI-LAH with respect to MP-LAH (30 mL [range 20-50] vs 50 mL [range 10-200], P < 0.05), with no statistically significant difference in terms of decrease of hemoglobin level (17 g/dL [range 2-24] vs 18 g/dL [range 5-28], P > 0.05). There were no significant differences between the two groups in terms of operative time (150 min [range 85-225] vs 145 min [range 100-220], P > 0.05), intraoperative injury, catheter retention time, time to exhausting, postoperative hospital stay. In all cases, no additional port incision was needed and no conversion to laparotomy was necessary in two groups. No patient had development of intraoperative or postoperative complications. After a follow-up of 2 months, no incisional hernia occurred in all patients. CONCLUSIONS: Single-incision laparoscopic hysterectomy using conventional instruments is a feasible and safe technique for patients with uterine size less than 12 weeks of pregnancy and no serious pelvic adhesion, requiring for more experienced skill of surgeons.

Stathmin 1 plays a role in endometrial decidualisation by regulating hypoxia inducible factor-1α and vascular endothelial growth factor during embryo implantation.

The aim of the present study was to explore the potential mechanism underlying stathmin 1 (Stmn1) regulation of embryo implantation, as a continuation of previous proteomic research. Adult healthy female mice were mated naturally with fertile males. Murine uterine tissue was collected during the peri-implantation period. Local expression of Stmn1 during embryo implantation was detected by immunohistochemistry (IHC), which showed that Stmn1 was extensively expressed in endometrial glandular epithelium, vascular endothelium, luminal epithelium and the underlying stromal cells at the implantation site on Day 5. The role of Stmn1 during embryo implantation was evaluated by transient knockdown of Stmn1 in vivo using short interference (si) RNA, and some associated factors including Akt, phosphorylated (p-) Akt, hypoxia-inducible factor (HIF)-1α, prolactin (PRL), insulin-like growth factor binding protein (IGFBP) 1 and vascular endothelial growth factor (VEGF) were examined by western blotting analysis and ELISA. The number of embryos implanted after Stmn1-siRNA infusion into the lumen of one uterine horn was lower than that with normal pregnancies (2.2±1.5 vs 8.6±0.5 respectively; P<0.05). The expression of VEGF, HIF-1α, p-Akt and the decidualisation biomarkers PRL and IGFBP 1 was upregulated at the implantation site on Day 5, but downregulated after Stmn1-siRNA infusion. These findings suggest that during embryo implantation, knockdown of Stmn1 suppresses decidualisation by inhibiting the expression of p-Akt, HIF-1α and VEGF, thus leading to impaired embryo implantation. These findings provide clues for understanding the complicated process of embryo implantation and the potential role of Stmn1 during embryo implantation.

Verteporfin, a suppressor of YAP-TEAD complex, presents promising antitumor properties on ovarian cancer.

Yes-associated protein (YAP) is a key transcriptional coactivator of Hippo pathway and has been shown to be an oncoprotein in ovarian cancer (OC). Verteporfin (VP), clinically used in photodynamic therapy for neovascular macular degeneration, has been recently proven to be a suppressor of YAP-TEAD complex and has shown potential in anticancer treatment. In this study, we aimed to explore the potential effect of VP in the treatment of OC. Our results showed that VP led to inhibition of proliferation in a time- and dose-dependent manner and to the suppression of migratory and invasive capacities of OC cells. Western blot and real-time polymerase chain reaction demonstrated that VP induced YAP cytoplasmic retention and deregulated inducible YAP and CCNs in OC cells. In vivo, VP exerted a significant effect on tumor growth in OVCAR8 xenograft mice, resulting in tumor nodules with lower average weight and reduced volume of gross ascites. In addition, VP treatment remarkably upregulated cytoplasmic YAP and phosphorylation YAP and downregulated CCN1 and CCN2, but exerted little effect on YAP-upstream components in Hippo pathway. In conclusion, our results suggested that VP may be a promising agent for OC, acting by suppressing YAP-TEAD complex.

Prognostic significance of TAZ expression in various cancers: a meta-analysis.

BACKGROUND: The overexpression of transcriptional coactivator with PDZ-binding motif (TAZ), a Hippo pathway effector, was detected in a variety of cancers. However, controversies remain in published studies on the prognostic value of TAZ expression in cancer. We performed a meta-analysis to demonstrate the prognostic significance of TAZ in overall survival (OS) and its association with clinicopathologic characteristics. METHODS: A systematic literature search was performed by using PubMed, EMBASE, and Web of Science databases for eligible studies investigating the association between TAZ and survival. After extracting data, we used hazard ratio (HR), odds ratio (OR) and 95% confidence intervals (95% CIs) for association evaluation, I (2) for heterogeneity across studies, and Egger's test and Begg's funnel plot for publication bias assessment. RESULTS: A total of 15 studies including 2,881 patients were analyzed. Pooled results showed that a high TAZ was significantly associated with poor OS (HR =1.82, 95% CI =1.58-2.11; I (2)=33%; P=0.11). Subgroup analysis indicated significant correlation between TAZ overexpression and OS in patients stratified by ethnicity, sample size, sample source, and staining location. Furthermore, TAZ overexpression was associated with worse OS in hepatocellular carcinoma (HR =2.26, 95% CI =1.43-3.57; P=0.49) and gastrointestinal cancers (HR =2.00, 95% CI =1.54-2.58; P=0.97), but not in non-small-cell lung cancer (HR =1.71, 95% CI =0.93-3.14; P=0.08). TAZ overexpression was also found to be significantly associated with some clinicopathologic characteristics, including TNM stage (OR =2.56, 95% CI =1.60-4.11; P=0.52), tumor differentiation (OR =3.08, 95% CI =1.25-7.63; P=0.01), and lymph node metastasis (OR =2.53, 95% CI =1.81-3.53; P=0.58). CONCLUSION: TAZ overexpression is not only a predictive factor of poor prognosis but also associated with advanced TNM stage, poor tumor differentiation, and lymph node metastasis.

Apolipoprotein A1 and heterogeneous nuclear ribonucleoprotein E1 implicated in the regulation of embryo implantation by inhibiting lipid peroxidation.

It is known that apolipoprotein A1 (apoA1) is a stimulator of endothelial nitric oxide synthase (eNOS), and that heterogeneous nuclear ribonucleoprotein E1 (hnRNP-E1)-containing RNP complexes is a key protector of basal stabilization of eNOS mRNA. Recently, we found that apoA1 and hnRNP-E1 were up-regulated during peri-implantation period, and the purpose of this study was to explore the roles of apoA1 and hnRNP-E1 during this period in the mouse. It was found that the up-regulation of apoA1 and hnRNP-E1 were dependent on the presence and status of blastocysts, on endometrial decidualization and on the progesterone and 17ß-oestradiol status. Knockdown of apoA1 or hnRNP-E1 both resulted in reduced numbers of embryo implantations and neonates (P < 0.01), and lipid peroxidation was found to be involved. On pregnancy day 5 eNOS expression and superoxidase dismutase (SOD) quantity were increased, and malondialdehyde (MDA) quantity was decreased at implantation sites. The knockdown of either apoA1 or hnRNP-E1 led to down-regulation of eNOS (P < 0.01) and to an increase in the quantity of MDA (P < 0.05), and a decrease in the amount of SOD (P < 0.01). These findings suggest that apoA1 and hnRNP-E1 may play roles in embryo implantation by inhibiting lipid peroxidation.

Immunotherapy of tumors with human telomerase reverse transcriptase immortalized human umbilical vein endothelial cells.

Human umbilical endothelial cells (HUVECs) have been proven to be effective in tumor anti-angiogenesis but the mechanism remained to be further demonstrated. The restricted ability of HUVECs to proliferate in vitro also limits their application on a large scale. In the present study, we immortalized HUVECs with hTERT genes by lentiviral infection and explored the antitumor immunity of hTERT-expressing HUVECs (HUVEC-TERTs). Results showed that HUVEC-TERTs maintained high telomere activity and expressed CD31, VEGFR-II and integrin α5. Passage-30 HUVEC-TERTs were able to form vascular tubes in vitro without showing signs of senescence. In vivo HUVEC-TERTs elicited antitumor immunity in mouse LL2 and CT26 models protectively and therapeutically. Both humoral and cellular immunity participated in the tumor anti-angiogenesis as HUVEC-neutralizing sera antibodies and HUVEC-specific CTL were detected. The subsets of activated spleen T lymphocytes included both CD4(+) T cells and CD8(+) T cells. Moreover, MDSCs and Tregs were decreased while T lymphocytes were aggregated in the tumor microenvironment. Collectively, the present study is the first to confirm the antitumor immunity of hTERT-immortalized HUVECs. Both anti-angiogenesis and tumor microenvironmental regulation participated in the antitumor activity. Transducing hTERT genes might be a new strategy to allow HUVECs to be applied on a large scale in cancer immunotherapy.

Mmu-miR-126a-3p plays a role in murine embryo implantation by regulating Itga11.

miR-126a-3p has been found to be specifically up-regulated in the process of murine embryo implantation. This study aimed to further clarify the role of miR-126a-3p in embryo implantation. The expression of miR-126a-3p in implantation sites was significantly higher than that in interimplantation sites (P = 0.009). Its expression dynamics in a series of models, including pseudopregnancy, delayed implantation and artificial decidualization, suggested that the induction of miR-126a-3p is dependent on hormonal signalling and decidualization of the endometrium. Bioinformatic analysis predicted and luciferase activity assay confirmed that Itga11, a member of the integrin family, was a target gene of miR-126a-3p. miR-126a-3p bound to the 3' untranslated region of Itga11 and regulated Itga11 by inhibiting mRNA translation and affecting mRNA stability. Transwell assay showed that miR-126a-3p promoted cell migratory and invasive capacity in vitro. Loss of function of miR-126a-3p significantly reduced the number of implantation sites in vivo (P = 0.013). Collectively, miR-126a-3p may play a major role in embryo implantation by regulating Itga11, possibly through impairing cell migratory and invasive capacity. These findings should contribute to a better understanding of the miRNA-based mechanism of embryo implantation.

Identification of stathmin 1 during peri-implantation period in mouse endometrium by a proteomics-based analysis.

In this work we aimed to identify the differentially expressed proteins and their potential roles during peri-implantation period through proteomics-based approach. Adult healthy female mice were mated naturally with fertile males to produce pregnancy. The models of pseudopregnancy, delayed implantation, and artificial decidualization were established. The protein profile between pre-implantation (D1) and implantation (D5) period was compared by two-dimensional electrophoresis (2-DE) and identified by mass spectrometry (MS). 2-DE yielded comparative images presenting over 500 protein spots in D1 and D5 mouse endometrium. 15 proteins were identified, of which stathmin 1, Apo-A1, hnRNP H3, transgelin 2 and arginase 1 were validated by western blotting. Stathmin 1 expression did not change in pseudopregnancy, but activation of implantation, or induction of decidualization increased it dramatically. Under non-pregnant status, progesterone alone or in combination with17ß-estradiol increased it dramatically. Our results clarified the protein profile in mouse endometrium during implantation. The specific expression profile of stathmin 1 suggested that it should be involved in implantation and serve as a potential regulator of this process. These findings may contribute to the better understanding of the molecules events during embryo implantation, and subsequently improve the ability to treat infertility.

MicroRNA-429 functions as a regulator of epithelial-mesenchymal transition by targeting Pcdh8 during murine embryo implantation.

STUDY QUESTION: What is the role of miR-429 in murine embryo implantation? SUMMARY ANSWER: miR-429 functions as a suppressor of epithelial-mesenchymal transition (EMT) during the process of embryo implantation by reverse regulation of Pcdh8. WHAT IS KNOWN ALREADY: MicroRNAs (miRNAs) may serve as promising regulators of embryo implantation. miR-429 was recently found to be down-regulated during embryo implantation period in a microarray analysis. STUDY DESIGN, SIZE, DURATION: The expression profile of miR-429 was clarified in a series of models, and the target gene was confirmed. The in vivo and in vitro effect of miR-429 on embryo implantation was examined. PARTICIPANTS/MATERIALS, SETTING, METHODS: Pregnancy was produced by natural mating between female C57BL6/J mice and male mice, and a series of models, including pseudopregnancy, delayed implantation and artificial decidualization, were established. The expression profile of miR-429 during the embryo implantation period was clarified in these models. Candidate target genes of miR-429 were predicted by bioinformatic analysis and tested by luciferase activity assay. The in vivo effects of miR-429 on embryo implantation were also examined. The in vitro effects of miR-429 on EMT were studied by examining migratory and invasive capacities by transwell assay and expression profiles of cadherin family members by western blotting and qRT-PCR. MAIN RESULTS AND THE ROLE OF CHANCE: The expression profile of miR-429 in animal models suggested its down-regulation should be dependent on the presence and status of blastocysts and on endometrial decidualization. The luciferase activity assay showed that Pcdh8, a member of cadherin gene family, was the target gene of miR-429, and miR-429 suppressed the expression of Pcdh8 mRNA and protein. Gain-of-function of miR-429 in vivo resulted in a significant reduction of the number of implantation sites, but had little effect on fertilization. Up-regulation of miR-429 in vitro led to suppression of mesenchymal marker genes Vim, Cdh2, Zeb1 and Zeb2, and activation of epithelial marker gene Cdh1, resulting in suppression of the migratory and invasive capacities of cells. miR-429 also partially abrogated TGF-beta-induced EMT. The dysregulated expression profiles of EMT markers during embryo implantation period could be partially reversed by gain-of-function of miR-429 in vivo. LIMITATIONS, REASONS FOR CAUTION: The association of miR-429 with other members of the miR-200 family in embryo implantation remains to be determined. The relationship between miR-429 and the cadherin family needs more intensive description and the detailed mechanism of miR-429 in regulating the cadherin family needs to be elucidated. WIDER IMPLICATIONS OF THE FINDINGS: Our findings indicate that miR-429 plays a major role in embryo implantation as a suppressor of EMT by targeting Pcdh8. This information could contribute to a better understanding of the mechanisms involved in the miRNA-mediated regulation of embryo implantation, and subsequently improve treatments for infertility. The findings are consistent with that from previous research of the other members in miR-200 family in embryo implantation and in the EMT. STUDY FUNDING/COMPETING INTERESTS: This study was supported by the Natural Science Foundation of China (Grant number: 81170592), and Special Fund from National Excellent Doctoral Dissertation (Grant number: 201079). There was no conflict of interest.

MicroRNA-451 plays a role in murine embryo implantation through targeting Ankrd46, as implicated by a microarray-based analysis.

OBJECTIVE: To determine the potential microRNA (miRNA) regulators of embryo implantation, as a continuation of genomic and proteomic research. DESIGN: Laboratory animal research. SETTING: University hospital laboratory. ANIMAL(S): Adult healthy female C57BL6/J mice (age 6-8 weeks, nonfertile, weighing 18-20 g each). INTERVENTION(S): Female mice were mated naturally with fertile males to produce pregnancy. Luminal epithelium was collected by laser-capture microdissection during the implantation period. Mouse models of pseudopregnancy, delayed implantation, and artificial decidualization were established. MAIN OUTCOME MEASURE(S): The miRNA profile in luminal epithelium was clarified by microarray analysis and validated by real-time reverse transcription polymerase chain reaction (qRT-PCR) in a series of models. Target genes were predicted and confirmed by luciferase activity assay. The role of miRNA in implantation was examined by loss-of-function and gain-of-function of miRNA in vitro and in vivo. RESULT(S): A total of 29 and 15 miRNAs were up- and down-regulated, respectively, during the implantation period; 11 of these miRNAs were validated by qRT-PCR. The profile of miR-451 was clarified in a series of models. A dual-luciferase activity assay showed that Ankrd46 was a target gene of miR-451. Loss-of-function by LV-miR-451 sponge or miR-451 inhibitor led to a reduced number of embryo implantations, but had little effect on fertilization. CONCLUSION(S): miR-451 was specifically up-regulated during the implantation period, and it may play a major role in embryo implantation by targeting Ankrd46.

Knockdown of cyclophilin A reverses paclitaxel resistance in human endometrial cancer cells via suppression of MAPK kinase pathways.

PURPOSE: Paclitaxel resistance remains to be a major obstacle to the chemotherapy of endometrial cancer. Using proteomic-based approach, we used to identify cyclophilin A (CypA) as a potential therapeutic target for endometrial cancer. As a natural continuation, this study aimed to reveal the correlation between CypA and paclitaxel resistance and evaluate the possibility of CypA as a therapeutic target for reversal of resistance. METHODS: Two paclitaxel-resistant endometrial cancer cell sublines HEC-1-B/TAX and AN3CA/TAX were generated, and expressions of CypA, P-gp, MRP-2 and survivin were demonstrated by Western blotting. CypA was knocked down by RNA interference, and the subsequent effects on the alteration of paclitaxel resistance were examined by MTT, flow cytometry and migratory/invasive transwell assays. MAPK kinases activities were examined by Western blotting. RESULTS: CypA knockdown led to significant inhibition of cell proliferation, induction of apoptosis and suppression of migratory/invasive capacity in HEC-1-B/TAX and AN3CA/TAX cells when exposed to paclitaxel. CypA knockdown led to reductions in total and phosphorylated MAPK kinases, including Akt, ERK1/2, p38 MAPK and JNK, in HEC-1-B/TAX cells. Furthermore, pretreatment with MAPK kinase inhibitors exhibited a synergistic effect in combination with CypA knockdown. CONCLUSIONS: These results demonstrated that CypA expression was up-regulated in paclitaxel-resistant cancer cells, and knockdown of CypA could reverse the paclitaxel resistance through, at least partly, suppression of MAPK kinase pathways, presenting a possibility of CypA serving as a therapeutic target to overcome paclitaxel resistance.