Prohibitin2/PHB2, Transcriptionally Regulated by GABPA, Inhibits Cell Growth via PRKN/Parkin-dependent Mitophagy in Endometriosis.

Endometriosis (EMS) is a common benign gynecological disease affecting women of reproductive age. It is characterized by abnormal growth of endometrial tissue outside the uterine cavity, resulting in chronic pelvic pain and infertility. Endometrial physiological and pathological processes are intimately connected to autophagy. Mitophagy is an essential selective mode that protects cells from metabolic stress and hypoxia. Mitochondrial autophagy mediated by prohibitin 2 (PHB2) is dependent on the PRKN/Parkin pathway and is involved in numerous human diseases. Uncertainty remains as to whether mitophagy regulation by PHB2 contributes to the occurrence and progression of EMS. This study aims to investigate the mechanism underlying the role of PHB2 in EMS. This study detected the protein and mRNA expression of PHB2 in ectopic and normal endometrial tissues of ovarian EMS, in addition to ectopic endometrial cell line 12Z and endometrial stromal cell line KC02-44D for gene overexpression or knockdown. Cell function experiments and mitochondrial function experiments were conducted to investigate the role of PHB2 in the endometrium. Bioinformatic analysis and experiments were also used to investigate the upstream transcription factors that influence PHB2 expression. PHB2 was downregulated in ectopic endometrium, and PHB2 overexpression inhibited cell proliferation, migration, and invasion and promoted apoptosis. The upregulation of mitophagy markers, including Parkin and LC3II/I, and the downregulation of autophagy degradation markers P62 and TOMM20 in EMS suggest that PHB2 may contribute to cell proliferation, migration, invasion, and apoptosis via PRKN/Parkin-mediated mitophagy. Analysis and validation of bioinformatics data revealed that the transcription factor GABPA binds directly to the PHB2 promoter region and controls the transcriptional expression of PHB2. This study investigated the role of PHB2 in the onset of EMS. It inhibits EMS growth via PRKN/Parkin-mediated mitophagy, and GABPA controls the transcriptional disorder of PHB2. This study's findings suggest a novel method for investigating the clinical potential of PHB2 in EMS.

Enhanced antifouling properties of marine antimicrobial peptides by PEGylation.

Covalent immobilisation of antimicrobial peptides (AMPs) on underwater surfaces to combat marine biofouling is of great interest as it is an efficient, broad-spectrum and environmentally friendly strategy. Similar to post-translational modifications of natural proteins, artificial modifications of antimicrobial peptides can introduce important impacts on their properties and functions. The present work revealed the enhanced effect of PEGylation on the antifouling properties of marine antimicrobial peptides (LWFYTMWH) through grafting the modified peptides on aluminium surfaces. PEG was coupled to the peptide by solid-phase peptide synthesis, and the PEGylated peptides were bioconjugated to the aluminium surfaces which was pre-treated by aryldiazonium salts to introduce carboxyl groups. The carboxy group has been activated through the reaction with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. The successful modification was confirmed via FT-IR and XPS. Interestingly, the PEGylated peptides modified surfaces could kill 90.0% Escherichia coli (Gram-negative) and 76.1% Bacillus sp. (Gram-positive), and showed better antifouling performance than the original peptides modified surfaces. Furthermore, molecular dynamics simulations showed PEGylation could enhance the ability of peptides to destroy membrane. The PEGylated peptides inserted into the membrane and induced the change in local curvature of membrane, leading to the rupture of membrane. The presence of PEG changed the antimicrobial peptides into more flexible conformations and the high hydrophilicity of PEG hindered the settlement of bacteria. These might be the two main working mechanisms for the increased antifouling efficiency of PEGylated peptides modified surface. This study provided a feasible modification strategy of antimicrobial peptides to enhance their antifouling properties.

FOXA1 can be modulated by HDAC3 in the progression of epithelial ovarian carcinoma.

FOXA1 is associated with malignant tumors, but the function of FOXA1 in EOC is unclear. HDAC3 can influence the proliferation, migration and invasion ability of EOC. In this study, we wanted to explore the function of FOXA1 in ovarian cancer and the relationship between HDAC3 and FOXA1.The expression of HDAC3 and FOXA1 was detected by immunohistochemical staining of primary lesions from 127 epithelial ovarian carcinoma patients. A proliferation assay, a Transwell assay, an apoptosis assay and animal experiments were used to assess the proliferation, invasion and apoptosis abilities of ovarian cancer cells before and after transfection with FOXA1. The relevance of the in vitro findings was confirmed in xenografts. The H-scores for FOXA1 and HDAC3 staining in FIGO stage III-IV were noticeably higher and predicted adverse clinical outcomes in patients with ovarian cancer. The expression level of HDAC3 was significantly correlated with the expression level of FOXA1. Invasion, proliferation and apoptosis capacity and tumor formation were decreased in the FOXA1-knockdown cells. Experiments in xenografts confirmed that HDAC3 mediated tumor formation. In conclusion, FOXA1 can be modulated by HDAC3 through the Wnt/ß-catenin signaling pathway, and FOXA1 plays essential roles in the proliferation, apoptosis and invasion of EOC cell lines and xenograft experiments.

MUC16 affects the biological functions of ovarian cancer cells and induces an antitumor immune response by activating dendritic cells.

BACKGROUND: Ovarian cancer is the 5th most common lethal gynecological malignancy with a 5-year survival rate of about 47% and a localized stage diagnosis of 15%, leading to about 125,000 global deaths each year. Therefore, it is urgent to explore novel and effective strategies for radical cure. METHODS: Short hairpin RNA targeting the Mucin16 (MUC16) gene was used to establish MUC16 knockdown in ovarian cancer cells. RT-PCR was performed to quantify the expression of MUC16 mRNA, and western blotting was performed to detect the expression of MUC16 and epithelial-mesenchymal transition-related proteins. Cell counting kit 8 (CCK8) wound healing and transwell assays were performed to assess cell proliferation and cell invasion. Flow cytometry was used to detect CD80-, CD83-, and CD86-expressing dendritic cells (DCs) and cytotoxic T lymphocytes (CTLs) activated by MUC16-pulsed DCs. RESULTS: In this study, we identified MUC16 as a novel target antigen for immunotherapy against ovarian cancer, which was significantly up regulated in ovarian cancer cells and high-grade ovarian serous adenocarcinoma tissues. MUC16 knockdown in Ovcar3 cells using short hairpin RNA targeting the MUC16 gene suppressed the proliferation of migration, invasion, epithelial-mesenchymal transition (EMT), and PI3K/Akt signaling pathway in Ovcar3 cells markedly. MUC16 significantly up-regulated CD80, CD83, and CD86 (mature makers) expression in DCs and T-cell transformation into CD8+ T-cells detected by Flow cytometry. CONCLUSIONS: For malignant ovarian cancer, MUC16 overexpression promoted cell proliferation, migration, and invasion via the PI3K/AKT signaling pathway. MUC16 pulsing mediated DC maturation and activated CTL response in vitro. Our study offers promising DC-based immunotherapy of considerable clinical value for patients with ovarian cancer.

Reproductive outcomes of cesarean scar pregnancies pretreated with methotrexate and uterine artery embolization prior to curettage.

OBJECTIVE: To evaluate the reproductive outcomes of cesarean scar pregnancy (CSP) pretreated with methotrexate (MTX) and uterine artery embolization (UAE) prior to curettage. MATERIALS AND METHODS: The medical records of patients with CSP who were pretreated with MTX and UAE prior to curettage in our institute from January 2013 to December 2015 were collected and retrospectively reviewed. RESULTS: A total of 53 patients were eligible for further analysis. Consecutive systemic MTX or a single dose of MTX was administered in 31 or 15 patients, respectively. The UAE procedure was uneventful, and no side effects occurred. The duration of the curettage operation was 21.4 ± 10.4 min, and the volume of blood loss was 23.5 ± 61.6 ml. The serum ß-HCG level returned to normal 36.1 ± 10.1 days after the date of initial MTX administration. Eight of 10 patients with a desire to have children became pregnant naturally. Two (25%) patients developed recurrent CSP during the first trimester. One patient underwent emergency cesarean delivery and hysterectomy due to placental implantation and sudden massive hemorrhage during delivery. A total of 6 live newborns were delivered. CONCLUSION: Pretreatment with MTX and UAE prior to curettage is safe and effective for the management of CSP. The reproductive outcomes are encouraging.

CK19 Promotes Ovarian Cancer Development by Impacting on Wnt/ß-Catenin Pathway.

BACKGROUND: Epithelial ovarian cancer (EOC) is the most common type of ovarian cancer and is the most lethal gynecologic malignancy. Cytokeratin 19 (CK19) is a small type I cytokeratin. The aim of this study is to explore the functional role of CK19 and its underlying mechanism in EOC. METHODS: The expression levels of CK19 in EOC tissues were identified by Western blotting and RT-PCR assay. Transwell assay and CCK-8 proliferation assay were used to assess the invasion, migration and proliferation abilities of overexpressed or knockdown CK19 of ovarian cancer cells. We also detected the related genes of Wnt/ß-catenin signal pathway, including ß-catenin, TCF7, LEF1, c-MYC and cyclin D1 in the transfected ovarian cancer cells by Western blotting and RT-PCR assay. RESULTS: The results demonstrated that CK19 was upregulated in EOC tissue. CK19 was verified to promote the invasion, proliferation and migration of ovarian cancer cells. Additionally, CK19 activates the Wnt/ß-catenin signaling pathway by upregulated ß-catenin, TCF7, LEF1, c-MYC and cyclin D1. CONCLUSIONS: In summary, this is the first study to investigate the role of CK19 in EOC. These findings provide a potential new therapeutic target for the clinical diagnosis and treatment of ovarian cancer.

HDAC3 positively regulates HE4 expression to promote ovarian carcinoma progression.

OBJECTIVE: To identify the relationship between Histone deacetylase 3 (HDAC3) and Human epididymis protein 4 (HE4) and to explore the mechanisms underlying their effects on the malignant behaviors of ovarian carcinoma cells. METHODS: The expression levels of HDAC3 in ovarian carcinoma tissues were identified by immunohistochemistry, Western blot and real-time PCR. A wound healing assay, a Transwell assay and a CCK8 proliferation assay were used to assess the proliferation, invasion and metastatic capacities of ovarian carcinoma cells before and after transfection and HDAC3 protein treatment. HDAC3 and HE4 protein expression level in epithelial ovarian tissues were detected by immunohistochemistry, and the relationship between them was examined. RESULTS: HE4 was identified as an HDAC3-interacting protein. HDAC3 promotes ovarian carcinoma cell proliferation, invasion and migration by increasing the expression of HE4. HE4 and HDAC3 expression levels were significantly higher in malignant epithelial ovarian tissues than they were in benign and normal epithelial ovarian tissues. HDAC3 gene interference downregulated the expression of the PI3K/AKT signaling pathway-associated molecules P-PI3K/PI3K and P-AKT/AKT. CONCLUSION: HDAC3 expression is higher in ovarian carcinoma and promotes ovarian carcinoma cell proliferation, invasion and migration. HDAC3 and HE4 binding activates the PI3K/AKT signaling pathway, enhances ovarian carcinoma and promotes ovarian carcinoma cell proliferation, invasion and migration. Therefore, inhibiting the relationship between HDAC3 and HE4 may therefore have potential therapeutic value in patients with ovarian carcinoma.

Construction and function of a fusion protein consisting of a single­chain variable fragment and RBP4.

Preeclampsia is one of the complications of pregnancy. Often presenting as new­onset hypertension and proteinuria after 20 weeks of gestation in a previously normotensive patient, it can progress rapidly to serious complications, and is a leading cause of maternal and perinatal mortality. The present study involved the construction of a fusion protein consisting of a single­chain antibody variable fragment (scFv) and the retinol­binding protein 4 (RBP4). and investigated the function of this protein. The MaxCodon™ Optimization Program (v13) was used to optimize the amino acid sequence of the scFv­RBP4 fusion protein and full­length splice primers were designed by Detai Bio Tech. The scFv­RBP4 gene was inserted into a proEM expression vector using double digestion, and the accuracy of the final expression vector was confirmed by restriction enzyme digestion and sequencing. The plasmid was transfected into DH5α competent cells and the plasmid was extracted from cells using a transfection reagent. The plasmid and scFv­RBP4 fusion protein were purified by nickel­iminodiacetic acid affinity chromatography. Cell proliferation was determined using the Cell Counting Kit­8 assay and cell invasion was measured using a Transwell invasion assay. The results from the digestion and sequencing showed that the scFv­RBP4 fusion protein was constructed correctly and that the purity of the target protein was >90%. The scFv­RBP4 fusion protein was stably expressed in 293T cells. The scFv­RBP4 fusion protein was extracted from the 293T cells and functional studies were carried out. The scFv­RBP4 fusion protein significantly increased the invasion, but not the proliferation, of HTR8/SVneo cells.

The security of radical trachelectomy in the treatment of IA-IIA cervical carcinoma requires further evaluation: updated meta-analysis and trial sequential analysis.

PURPOSE: The aim of this study was to assess the security of radical trachelectomy (RT) in the treatment of IA-IIA cervical carcinoma and conducted a new survey based upon the results of previous researches. METHODS: The PMC, PubMed, Web of Science, Cochrane and EMBASE databases were retrieved to collect prospective clinical controlled trials (CCTs) published from 1984 to 2018. The oncologic outcomes were evaluated by meta-analysis, trial sequence analysis (TSA) and statistical analysis. RESULTS: Five prospective CCTs were collected in this study. The recurrence rate and mortality of RT was similar to that of radical hysterectomy (RH), which was consistent with the oncologic outcomes of meta-analysis and TSA. Patients with tumors 2-4 cm in diameter were more likely to receive RH, which may be a potential factor in the higher rate of adjuvant chemotherapy in the this group, and RH was significantly associated with the risk of intraoperative blood transfusion. It is notable that considerable negative margin was achieved by radical abdominal trachelectomy (RAT), and the clinical effect of RAT was slightly better than that of radical vaginal trachelectomy (RVT). However, the TSA results showed that the cumulative cases were not up to the required sample size to obtain the true negative or positive results. CONCLUSIONS: It is safe and effective for early-stage patients with cervical cancer whose lesions are less than 2 cm to receive RVT. For those patients with lesions 2-4 cm who desire fertility preservation and without any evidence of infertility, RAT can be a feasible alternative to RH under fully informed consent. However, more CCTs with larger sample size are still required for further validation.

RBP4 regulates trophoblastic cell proliferation and invasion via the PI3K/AKT signaling pathway.

Insufficient trophoblast invasion is associated with preeclampsia (PE) development. Retinol-binding protein 4 (RBP4) is important for regulating cell differentiation, migration and invasion. The aim of the present study was to determine RBP4 expression and function in the human placenta and to examine the underlying mechanisms. In the present study, RBP4 expression was determined in serum samples from 35 pregnant women with PE and 30 healthy pregnant women using enzyme-linked immunosorbent assays. Cell proliferation was assessed by Cell Counting Kit-8 assays, and cell invasion was examined with transwell assays. RBP4 concentrations were significantly lower in the PE group when compared with the control group. RBP4 overexpression enhanced HTR8/SVneo cell proliferation and invasion, and the levels of phosphorylated (p-) phosphoinositide 3-kinase (PI3K) and p-protein kinase B (AKT) in HTR8/SVneo cells. RBP4 knockdown significantly inhibited HTR8/SVneo cell proliferation and invasion, and repressed the expression of matrix metalloproteinases. In addition, RBP4 knockdown significantly reduced the levels of p-PI3K and p-AKT in HTR8/SVneo cells. Taken together, the results of the present study demonstrated that RBP4 overexpression increased HTR8/SVneo cell proliferation and invasion by suppressing PI3K/AKT signaling and RBP4 knockdown induced the opposite effects.

The safety of fertility preservation for microinvasive cervical adenocarcinoma: a meta-analysis and trial sequential analysis.

PURPOSE: This study sought to evaluate the safety of conservative treatment in the management of patients with microinvasive cervical adenocarcinoma. METHODS: The PubMed, PMC, EMBASE, Web of Science and Cochrane databases were searched to collect correlational studies published in English between January 1949 and May 2018. Series reports that evaluating the oncological prognoses of patients with microinvasive cervical adenocarcinoma who were treated with fertility-sparing surgery versus hysterectomy were pooled for meta-analysis and trial sequential analysis. RESULTS: A total of 8 articles with 1256 patients were collected, including 7 retrospective reviews and 1 prospective study. Only one (0.08%) patient had parametrial involvement. Positive margins of surgical specimens were identified in 6 patients (2.2%). Lymph node metastasis was found in 5 patients (0.4%). The progression-free survival and overall survival rates were 99.3 and 98.2%. Fertility-sparing surgery had no adverse impact on recurrence or survival (P = 0.524 and 0.485, respectively). Regarding potential selection bias, significantly more patients with stage IA2 tumors than those with stage IA1 disease were treated with hysterectomy (P < 0.001). The trial sequential analysis indicated that the cumulative number of patients failed to meet the required sample size (number of patients). CONCLUSIONS: The prognosis for patients with microinvasive cervical adenocarcinoma is excellent. Fertility preservation is at least appropriate for young women with stage IA1 adenocarcinoma. Further studies are still warranted to evaluate the safety of this procedure in managing patients with microinvasive cervical adenocarcinoma.

Oncological and reproductive outcomes of adenocarcinoma in situ of the cervix managed with the loop electrosurgical excision procedure.

BACKGROUND: The standard treatment for cervical adenocarcinoma in situ (AIS) is hysterectomy, which is a more aggressive treatment than that used for squamous intraepithelial lesions. Several previous studies have primarily demonstrated that the loop electrosurgical excision procedure (LEEP) is as safe and effective as cold knife cone (CKC) biopsy when AIS is unexpectedly found in a loop excision. This study evaluated the safety of LEEP as the initial treatment for patients with AIS who were strictly selected and evaluated before and after loop resection. METHODS: The oncological and reproductive outcomes of a series of AIS patients who underwent LEEP as the initial treatment between February 2006 and December 2016 were retrospectively evaluated. RESULTS: A total of 44 women were eligible for analysis. The mean age at diagnosis was 36.1 years, and 14 patients were nulliparous. Multiple lesions were identified in 4 (9.1%) patients. Either hysterectomy (6 patients) or repeat cone biopsies (3 patients) were performed in 8 of the 10 patients who presented positive or not evaluable surgical resection margins (SMs) on the initial LEEP specimens. Residual disease was detected in two patients. All patients were closely followed for a mean of 36.9 months via human papillomavirus testing, PAP smears, colposcopy, and endocervical curettage when necessary. No recurrences were detected. Of the 16 patients who desired to become pregnant, 8 (50%) successfully conceived, and the full-term live birth rate was 83.3% among this subgroup. CONCLUSIONS: LEEP with negative SMs was a safe and feasible fertility-sparing surgical procedure for patients with AIS, and the obstetric outcome was satisfactory. However, long-term follow-up is mandatory.

N-myc Downstream-Regulated Gene 1 and Endometriosis: A Minireview.

NDRG1 (N-myc downstream-regulated gene 1) was previously considered to be a differentiation-related gene. However, many other functions of NDRG1 have since been identified, including proliferation, migration, invasion, and vascularization of tumor cells. Currently regarded as a tumor suppressor in most studies, NDRG1 is abundant in prostate, brain, kidney, placental, and intestinal tissues. It is expressed in normal endometrium, with higher expression occurring in the secretory phase. NDRG1 was first identified as an inhibitor of signaling pathways associated with the pathology of endometriosis. The NDRG1 protein regulation of endometriosis is assumed to be associated with several important pathways. This review summarizes the relationship between NDRG1 and endometriosis, focusing on the potential function of NDRG1 in endometriosis through signaling pathways and discusses the additional research that is required for future studies.

The safety of fertility and ipsilateral ovary procedures for borderline ovarian tumors.

OBJECTIVE: To explore the optimal treatment options for women with borderline ovarian tumors (BOTs). MATERIALS AND METHODS: The medical records of consecutive patients with BOTs in two academic institutions were retrospectively collected. The pertinent data, including clinicopathological characteristics and, treatment and prognostic information were evaluated. RESULTS: A total of 281 cases of BOTs were included in this analysis. For the entire series, the 5- year disease-free survival (DFS) and overall survival (OS) rates were 91.8% and 98.5%, respectively. In the multivariate analysis, reservation of the ipsilateral ovary (HR: 0.104 [95% CI, 0.036-0.304], p = 0.000) and FIGO stage II-III (HR: 6.811 [95% CI, 2.700-17.181], p = 0.000) were the independent risk factors for recurrence. Ovarian surface involvement (HR: 64.996 [95% CI, 4.054-1041.941], p = 0.003) was the only independent prognostic factor for OS. Lymphadenectomy and adjunct chemotherapy had no significant impact on patients' recurrence and survival (recurrence: p = 0.332 and 0.290, respectively, survival: p = 0.896 and 0.216, respectively). CONCLUSIONS: Fertility-sparing surgery with healthy ovarian preservation seems safe and feasible for young women who prefer fertility-sparing treatment. Ovarian cystectomy to conserve the affected ovary/ovaries without ovarian surface involvement may be cautiously performed under fully informed consent for young women with bilateral BOTs who strongly prefer fertility-sparing treatment and have no evidence of infertility. However, long-term follow-up is necessary due to the relapse susceptibility of the ovary.