Differentially expressed EREG and SPP1 are independent prognostic markers in cervical squamous cell carcinoma.

AIMS: Cervical squamous cell carcinoma (SCC) is one of the most frequent malignancies of the female reproductive system. The malignant mechanism of SCC has not been totally clarified. We aimed to discover a list of differentially expressed genes (DEGs) to identify the malignant mechanism of cervical SCC. METHOD: Three expression chips (GSE7803, GSE9750, and GSE64217) were downloaded from gene expression omnibus (GEO) datasets. After standardization, 50 cervical SCC tumor tissues and 33 normal cervical tissues (NCTs) were included for DEGs and clustering analysis. RobustRankAggreg (RRA) algorithm was used to extract the overlapping DEGs. Gene function and signaling pathway analysis was implemented based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway databases. Protein-protein interaction (PPI) analysis and prognostic analysis were also carried out to identify the DEGs as prognostic markers for cervical SCC. RESULTS: Totally 100 DEGs were obtained from GSE7803, 319 DEGs from GSE9750, and 1639 DEGs from GSE64217. RRA analysis uncovered 17 upregulated DEGs and 25 downregulated DEGs. GO and KEGG analysis showed DEGs were involved in the mediation of extracellular functions, cell-cell interactions, and cell metabolism. PPI network showed a close interaction among the integrated DEGs. Prognostic analysis showed gene secreted phosphoprotein 1 (SPP1) and epiregulin (EREG) genes were independent prognostic predictors of cervical SCC. CONCLUSION: The gene expression profile was changed in cervical SCC tumor tissues compared to NCTs. SPP1 and EREG were postulated as prognostic markers for cervical SCC, which might be potential targets for clinical therapy of cervical SCC.

Resistance to bevacizumab in ovarian cancer SKOV3 xenograft due to EphB4 overexpression.

AIM OF STUDY: Bevacizumab (BV) is broadly used to treat a number of cancers; however, BV resistance mechanisms and strategies to overcome this resistance are yet to be determined. MATERIALS AND METHODS: In this study, we used ovarian xenograft model to evaluate the underlying resistance mechanisms of BV in ovarian cancer treatment. RESULTS: Our results showed that EphB4 was overexpressed in BV-resistant xenograft models instead of other common receptor tyrosine kinases. In addition, when coadministrated with EphB4 blocker NVP-BHG712, the antitumor effect of BV was significantly enhanced in the resistant model, further confirmed the role of EphB4 in BV-resistant ovarian cancer. These results indicate that NVP-BHG712 reverses EphB4 overexpression-mediated resistance to BV. CONCLUSION: These findings represent a guide for the design of future medication strategy and may be useful in guiding the use of BV in combination with NVP-BHG712 in patients with resistance or intolerance ovarian cancer.

Post hysteroscopic progesterone hormone therapy in the treatment of endometrial polyps.

OBJECTIVE: To find out the clinical effects of post hysteroscopic progesterone hormone therapy in the treatment of endometrial polyps in terms of clinical outcome and the expression of endometrial Vascular Endothelial Growth Factor (VEGF). METHODS: Ninety-eight patients who were confirmed as endometrial polyp in the hospital from April 2014 and December 2016 were selected and divided into treatment group and a control group using random number table, 49 in each group. Patients in both groups were given hysteroscopic operation. Patients in the treatment group were treated by progesterone hormone drugs after hysteroscopic operation, while patients in the control group were not given progesterone hormone. The changes of menstrual blood volume, menstrual cycle and expression of VEGF were compared between the two groups after treatment, and the recurrence condition, thickness of endometrium and hemoglobin were followed up one year after treatment. RESULTS: The pictorial blood loss assessment chart (PBAC) scores of patients in the two groups had no significant difference before treatment (P>0.05); but the score of the treatment group was much lower than that of the control group. The improvement rate of menstrual cycle of the treatment group was much higher than that of the control group, and the difference had statistical significance (P<0.05). Compared to before treatment, the serum VEGF level of the patients in both groups had a remarkable decline in the 1st, 3rd and 6th month after treatment, and the difference had statistical significance (P<0.05). The difference of the serum VEGF level between the two groups in the 1st and 3rd month after treatment had no statistical significance (P>0.05). The serum VEGF level of the treatment group was notably lower than that of the control group six months after treatment, and the difference had statistical significance (P<0.05). The follow-up results demonstrated that the treatment group had smaller thickness of endometrium and higher level of hemoglobin compared to the control group, and the recurrence rate of the treatment group was lower than that of the control group (P<0.05). CONCLUSION: Post hysteroscopic progesterone hormone therapy has favorable clinical effect in treating endometrial polyps as it can effectively prevent the recurrence of endometrial polyps, relieve the level of hemoglobin and reduce endometrial thickness.

Asiatic acid inhibits LPS-induced inflammatory response in endometrial epithelial cells.

Accumulating evidence indicates that asiatic acid, a natural triterpene isolated from Centella asiatica, has anti-inflammatory activity. However, the anti-inflammatory effects of asiatic acid on LPS-stimulated endometrial epithelial cells and the involved molecular pathways have not been completely elucidated. In the present study, we evaluated the effects of asiatic acid on LPS-induced inflammatory response in endometrial epithelial cells. Mouse endometrial epithelial cells were treated with asiatic acid and stimulated with LPS. ELISA was performed to measure the levels of inflammatory cytokines TNF-α, IL-1ß, and PGE2. Western blot analysis was used to test the expression of PPARγ and NF-κB. The results showed that LPS-induced inflammatory mediators TNF-α, IL-1ß, NO, and PGE2 were significantly inhibited by asiatic acid. Furthermore, LPS-induced TLR4 expression and NF-κB activation were concentration-dependently suppressed by asiatic acid. In addition, asiatic acid was found to increase the expression of PPARγ in a concentration-dependently manner. The inhibition of asiatic acid on inflammatory mediators production were prevented by PPARγ inhibitor, GW9662. Taken together, these results showed that asiatic acid exhibited its anti-inflammatory effects in endometrial epithelial cells by activating PPARγ.

Suppressive efficiency of RASSF1A in endometrial carcinoma via inhabiting estrogen receptor alpha expression and ERK pathway activation.

INTRODUCTION: Known as a tumor suppressor, the Ras association domain family 1 isoform A (RASSF1A) is implicated in many human cancers, such as endometrial carcinoma. There is little known about the tumor inhibitive effects of RASSF1A on endometrial carcinoma. The present study was designed to investigate the role of RASSF1A in HEC-1-A cells and to explore its potential mechanisms. MATERIALS AND METHODS: In this study, overexpression of RASSF1A was established by transfection the recombinant adenoviral RASSF1A in HEC-1-A cells. Cells viability was assessed by MTT assay and the apoptosis was analyzed using flow cytometry. Cell migration and invasion were measured in Transwell assay. The levels of ERα and PELP1 protein and extracellular regulated protein kinase (ERK) pathway activation were detected by Western blot. RESULTS: RASSF1A over-expression could significantly inhibit the proliferation, migration and invasion of the HEC-1-A cells in transfection with RASSF1A group compared to that in transfection with control group, also induced apoptosis and suppressed the tumor growth after injection in nude mice. Moreover, overexpression of RASSF1A could inhibit the ERK signal pathway activation and decrease the ERα and PELP1 expression. CONCLUSION: Tumor suppressive efficiency of RASSF1A is exerted through the regulation of ERK pathway activation, ERα and PELP1 expression.

Identification of key genes in endometrioid endometrial adenocarcinoma via TCGA database.

BACKGROUND: Understanding the molecular mechanisms is important in development and therapy of endometrioid endometrial adenocarcinoma. OBJECTIVE: To identify key genes in endometrioid endometrial adenocarcinoma. METHODS: The data of mRNA, miRNA and DNA methylation were downloaded from The Cancer Genome Atlas (TCGA) database and differential analysis was performed. Then, bioinformatic analysis was used to explore the regulatory mechanisms of miRNA and DNA methylation on gene expression. The regulatory network between differentially expressed miRNAs and target genes was established. Finally, the quantitative RT-PCR was applied to validate the bioinformatics results. RESULTS: We obtained biological omics data of 381 patients with endometrioid endometrial adenocarcinoma from TCGA data portal. After data processing, up to 2068 DEGs and 69 differentially expressed miRNAs were identified. Prediction and correlation analysis revealed that 175 DEGs that were not only the target genes but also negatively correlated with the screened differentially expressed miRNAs. After the integrated analysis of differentially methylated CpG islands and DEGs, 16 related genes were obtained. The quantitative RT-PCR results were roughly consistent with the bioinformatics analysis. CONCLUSIONS: The altered DEGs (ZEB1, ZEB2, TIMP2, TCF4, CYP1B1, PITX1, PITX2, ZNF154 and TSPYL5) may be involved in tumor differentiation of endometrioid endometrial adenocarcinoma and could be used as potential therapeutic targets for the disease.

5-Aminolevulinic acid photodynamic therapy in human cervical cancer via the activation of microRNA-143 and suppression of the Bcl-2/Bax signaling pathway.

5-Aminolevulinic acid photodynamic therapy (ALA­PDT) is a method using a photosensitizer and light radiation for disease treatment, and is currently used for the treatment of skin cancers, precancerous lesions and viral warts. The present study aimed to investigate the effect of ALA­PDT on human cervical cancer through the regulation of microRNA­143 (miR­143) and the Bcl-2/Bax signaling pathway. The results demonstrated that ALA­PDT reduced proliferation, increased cytotoxicity and induced apoptosis in HeLa human cervical carcinoma cells. Reverse transcription­quantitative polymerase chain reaction analysis demonstrated that the expression levels of miR­143 were increased following ALA­PDT treatment. Western blotting indicated that the expression levels of Bcl­2 and Bax were significantly reduced and increased, respectively, by ALA­PDT treatment. In addition, upregulation of miR­143 expression reduced Bcl­2 expression and increased Bax expression in HeLa cells. However, downregulation of miR­143 expression inhibited the effect of ALA­PDT on Bcl-2/Bax protein expression. In conclusion, the current study demonstrated that ALA­PDT affected human cervical cancer via the activation of miR-143 and the suppression of the Bcl-2/Bax signaling pathway.

Systematic studies of Pickering emulsions stabilized by uniform-sized PLGA particles: preparation and stabilization mechanism.

Pickering emulsions stabilized by solid particles have been widely studied in the past decades due to improved stability and reduced use of small molecular surfactants. Recently, the application of Pickering emulsions in pharmaceutics has been attracting increasing attention but very limited practical use has been demonstrated, because most of the investigated particles possess poor biodegradability, which is inappropriate in pharmaceutics. Some reported biodegradable particles were too hydrophilic to stabilize emulsions, which needs further particle modification or additional surfactants. Fortunately, biodegradable poly(d,l-lactic-co-glycolic acid) (PLGA) with tunable hydrophilicity makes itself a promising material to prepare Pickering emulsions. However, the mechanism of emulsion stabilization still remains unknown. Moreover, fabrication of large amounts of uniform-sized and size-controlled PLGA particles by traditional methods is very difficult, which further increases the difficulty to perform the research. In the present study, we applied Shirasu Porous Glass (SPG) premix membrane emulsification to solve this problem. The stabilization mechanism of Pickering emulsions stabilized by PLGA particles was systematically studied for the first time. The factors including oil type, particle properties, concentration, molecular weight (Mw) and oil-water volume ratio were analyzed through particle wettability and interfacial influence. We found that octanol was an appropriate oil type, and its small particle size, high particle concentration and high Mw were favorable for emulsion stability. By changing the oil-water volume ratio, stable emulsions were also readily achieved. These studies proved that Pickering emulsions stabilized by PLGA particles had wide potential applications in pharmaceutics and tissue engineering.

Preparation of uniform-sized colloidosomes based on chitosan-coated alginate particles and its application for oral insulin delivery.

Colloidosomes are microcapsules that consist of a hollow core coated by a shell composed of self-assembled colloidal particles. In recent years, they have attracted significant attention as potential vehicles for the controlled delivery of active ingredients. A key challenge in such applications is the production of uniform-sized colloidosomes for the encapsulation of active ingredients such that the actual delivery amount can be well controlled. Based on our previous study on an ethyl acetate-in-water emulsion stabilized by chitosan-coated alginate particles, we further prepared monodisperse colloidosomes with a high yield and low permeability by combining the premix membrane emulsification technique and polymer deposition method. The potential application of the formed colloidosomes as oral insulin delivery vehicles was investigated. Our results revealed that in comparison to conventional PLGA microspheres, the formulated colloidosomes showed high drug encapsulation efficiency (up to 96.7%) and an obvious pH sensitive release profile. Moreover, in animal testing, the colloidosomes formulation achieved a long-term hypoglycemic effect up to 6 h, which confirmed its application as an oral drug delivery system.

B7-H4 expression promotes tumorigenesis in ovarian cancer.

INTRODUCTION: It has been previously shown that B7-H4, one of the B7 family members that serve as negative regulators of T cell function, has altered expression levels in a variety of cancers, overexpression of B7-H4 promotes cellular transformation. However, there is still lack of adequate evidence to establish a direct connection between B7-H4 expression and malignant transformation. METHODS: Herein, we constructed pE-green fluorescent protein-N1/B7-H4 mammalian expression vector and transfected into B7-H4-negative human ovarian cancer cell line SKOV3. Cellular proliferation, apoptosis, adhesion, motility, and invasion were examined in vitro. Cells injected subcutaneously into severe combined immunodeficient mouse were analyzed for the possible functions of B7-H4 in ovarian tumorigenesis in vivo. RESULTS: Fluorescence microscopy studies confirmed that the B7-H4-green fluorescent protein localizes in the cytoplasm of SKOV3/B7-H4 cells, whereas green fluorescent protein is uniformly distributed throughout the cell. B7-H4 promoted cellular proliferation rate and increased cell adhesion, migration, and invasion. In addition, SKOV3 cells expressing B7-H4 gained growth advantage in the xenograft model in vivo. CONCLUSIONS: These studies demonstrate that B7-H4 directly promotes malignant transformation of ovarian cancer cell line, and provides a potential therapeutic strategy for targeting B7-H4 to inhibit progression of human ovarian cancers.

Altered expression of DACH1 and cyclin D1 in endometrial cancer.

The altered expression of human DACH1, a Drosophila Dachshund homolog, has been associated with tumor progression and metastasis. DACH1 inhibits breast cancer cellular proliferation via cyclin D1. Endometrial cancer is the third most common cancer in women and broad screening for DACH1 expression will further our understanding of this disease. Herein, we screened 126 hysterectomy specimens for DACH1 expression and evaluated the correlation between DACH1 levels and several clinical parameters. Decreased DACH1 expression was significantly correlated with FIGO surgical stages (Stage I-II vs. stage III-IV, p = 0.017), peritoneal cytology (p = 0.044), lymph node positivity (p = 0.035) and histological type (p = 0.007), but not histological grade, depth of myometrial and patient age. Immunostaining was also conducted to examine the expression of cyclin D1, estrogen receptor alpha (ERalpha) and progesterone receptor (PR) in these specimens. Multivariate analysis using the stepwise Cox proportional hazard model showed that FIGO surgical stage, histological grade, lymph node metastasis, and PR expression were correlated with poor survival. Despite the fact that univariate analysis demonstrated that DACH1 positivity is associated with increased 5-year survival in all patients (p = 0.037), decreased expression of DACH1 had no significant value as an independent prognostic factor in predicting survival in endometrial cancers. Our results suggested that loss of DACH1 expression might be involved in endometrial cancer progression.

[Combination chemotherapy with etoposide and cisplatin for high-risk, chemorefractory and recurrent gestational trophoblastic neoplasia].

OBJECTIVE: To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin (EP) regimen on the patients with high-risk, chemorefractory and recurrent gestational trophoblastic neoplasia (GTN). METHODS: Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively, 25 of 39 patients were of high-risk, 9 patients were chemorefractory and 5 patients were recurrent. All 39 patients were administrated with EP regimen, and 10 patients were assisted with surgery. All the patients were followed up. Clinical response, toxicity, the occurrence of secondary tumors of all patients, and the fertility of 30 patients whose fertility function was preserved were investigated. RESULTS: Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen. The average number of courses for each patient was 5.7. The total complete remission rate of the regimen was 74% (29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6. Nineteen patients achieved complete remission and 6 patients showed drug-resistant. The complete remission rate of the high-risk group was 76% (19/25). Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1. Six patients achieved complete remission and 3 patients showed drug-resistant again. The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4. Four patients achieved complete remission, 1 patient showed drug-resistance and died. Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen, but the bone marrow toxicity was slight and no grade IV side effect occurred. No fatal effect was found. Eight of 30 patients whose fertility fuction was preserved had become pregnant after recovery, with a total of 8 pregnancies. Among them, 2 were terminated by induced abortion, and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation. All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors. CONCLUSION: The EP regimen is effective and safe for the treatment of high-risk, chemorefractory and recurrent GTN.