Genetic polymorphism of IL-17 influences susceptibility to recurrent pregnancy loss in a Chinese population.

The current research aims to investigate the relationship between Interleukin-17 (IL-17) polymorphism and the risk of recurrent pregnancy loss (RPL) within a Chinese population. Totally, 120 patients with RPL were selected and enrolled as the experiment group. Additionally, 210 healthy individuals undergoing routine physical examinations during the same period served as the control group. The IL-17 gene polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism method. The IL-17 rs2275913 polymorphism exhibited 3 genotypes: GG, GA, and AA. Significant associations were observed with the AA genotype and A allele (all P < .05), indicating women with the AA genotype were 2.06 times more likely to experience RPL compared to those with the GG genotype. Similarly, women carrying the A allele faced a 1.63 times higher risk of RPL than those with the G allele. Regarding the IL-17 rs763780 polymorphism, which also presented 3 genotypes (TT, TC, CC), significant associations were noted for the CC genotype and C allele (all P < .05). Women with the CC genotype had a 1.84 times greater risk of suffering from RPL compared to those with the TT genotype, and those with the C allele were 1.51 times more likely to experience RPL than those with the T allele. The IL-17 rs2275913 and rs763780 polymorphisms contribute an increased risk to RPL in the Chinese population. Further studies, with larger sample sizes and more rigorous designs, are necessary to validate or replicate our current results.

A quadruple-stimuli responsive supramolecular hydrogel constructed from a poly(acrylic acid) derivative and ß-cyclodextrin dimer.

The fabrication of stimulus-responsive supramolecular hydrogels as smart materials has attracted much attention in recent years. However, the multi-stimuli responsiveness often requires complicated chemical synthesis and rational molecular design. Herein, a quadruple-stimuli responsive supramolecular hydrogel was designed through the host-guest interaction between a ß-CD dimer and a methoxy-azobenzene (mAzo) and ferrocene (Fc) grafted poly(acrylic acid) derivative, as well as through the electrostatic interaction of negatively charged carboxyl side groups. Owing to the dynamic properties of the host-guest and electrostatic interactions, reversible sol-gel transition can be triggered by various stimuli, including temperature, light irradiations, pH changes and chemical redox reagents. As a result, the release of rhodamine B loaded in the hydrogel can be accelerated by green light irradiation, oxidizing agents and low pH, demonstrating potential applications in biomedical materials.

Pyroptosis is involved in the immune microenvironment regulation of unexplained recurrent miscarriage.

Unexplained recurrent miscarriage (URM) is a common pregnancy complication with few effective therapies. Moreover, little is known regarding the role of pyroptosis in the regulation of the URM immune microenvironment. To address this issue, gene expression profiles of publicly available placental datasets GSE22490 and GSE76862 were downloaded from the Gene Expression Omnibus database. Pyroptosis-related differentially expressed genes were identified and a total of 16 differentially expressed genes associated with pyroptosis were detected, among which 1 was upregulated and 15 were downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that the functionally enriched modules and pathways of these genes are closely related to immune and inflammatory responses. Four hub genes were identified: BTK, TLR8, NLRC4, and TNFSF13B. BTK, TLR8, and TNFSF13B were highly connected with immune cells, according to the correlation analysis of four hub genes and 20 different types of immune cells (p < 0.05). The four hub genes were used as research objects to construct the interaction networks. Chorionic villus tissue was used for quantitative real-time polymerase chain reaction and western blot to confirm the expression levels of hub genes, and the results showed that the expression of the four hub genes was significantly decreased in the chorionic villus tissue in the URM group. Collectively, the present study indicates that perhaps pyroptosis is essential to the diversity and complexity of the URM immune microenvironment, and provides a theoretical basis and research ideas for subsequent target gene verification and mechanism research.

Blocking PARP activity with the inhibitor veliparib enhances radiotherapy sensitivity in endometrial carcinoma.

OBJECTIVE: Our study aimed to investigate the potential clinical utility of a poly(ADP-ribose) polymerase (PARP) inhibitor, veliparib (ABT-888), as a radiosensitizer in the medication of endometrial carcinoma (EC). METHODS: Human Ishikawa endometrial adenocarcinoma cells were treated with veliparib, radiotherapy (RT), or combination treatment. The viabilities, radiosensitivity enhancement ratio (sensitizer enhancement ratio (SER), and apoptosis of Ishikawa cells were, respectively, evaluated by Cell Counting Kit-8 (CCK-8), colony formation experiment, and flow cytometry. The tumor growth was assessed by xenograft mice models. Western blot assay investigated the expression of DNA damage and apoptosis-related proteins in vivo and in vitro. RESULTS: Cell Counting Kit-8 revealed that the 10% inhibition concentration (IC10 ) and 50% inhibition concentration (IC50 ) values of veliparib-treated Ishikawa cells were 1.7 and 133.5 µM, respectively. The SER of veliparib combined with RT was 1.229 in vitro. Flow cytometry analysis results indicated that the apoptosis rate of the veliparib + RT group was markedly higher than that of the RT group in vitro (p < 0.05). Furthermore, in vivo data revealed that veliparib + RT treatment significantly decreased tumor growth compared with single treatments of veliparib or RT and with the control group (p < 0.05). Then western blot confirmed the levels of anti-phospho-histone (γH2AX), caspase-3, and B-cell lymphoma 2 (Bcl-2) associated protein X (Bax) were significantly higher in the veliparib + RT group, while the level of Bcl-2 was lower compared with that of the RT group (p < 0.05), both in vivo and in vitro. CONCLUSION: Our results indicate that veliparib in combination with RT markedly improved the therapeutic efficiency in human endometrial carcinoma.

Light regulation and long-lived stability of RGB colors in cholesteric liquid crystal physical gels via a mixing strategy.

Photo-responsive cholesteric liquid crystals (CLCs) have attracted much attention due to the dynamic tunability of their unique helical superstructure. However, it is still a challenge to endow the mechanical properties and to regulate the reflection colors at the same time. In this work, a simple strategy is developed for the construction of thermo-responsive CLC physical gels via the direct mixing of photo-responsive dopants and a gelator with nematic LCs. The reflection colors of CLCs and the mechanical properties of gels can be independently regulated due to the separation of the photo-responsive chiral group from the gelator. In addition, the CLC reflection colors can be regulated via visible light in the range of RGB with long-lived thermal stability. Finally, the information storage properties of this kind of CLC gel have been investigated.

[Effect of Indirubin and Sheng-Xue-Xiao-Ban Capsule (SXXBC) on Promoting Peripheral Platelet in ITP Model Mice].

OBJECTIVE: To compare the effect of Sheng-Xue-Xiao-Ban Capsule (SXXBC) and indirubin to the peripheral platelets of the Idiopathic thrombocytopenic purpura (ITP) model mouse. METHODS: The ITP mouse model was established by the method of passive immunization. SXXBC and indirubin were used for intervention treatment. Then the hemorrhagic phenomena of ITP mice were observed and the numbers of peripheral platelets, hemoglobin and white blood cells, bone marrow megakaryocytes and their classification and coagulation function were detected and compared. RESULTS: The improvement rate of hemorrhage in SXXBC group was 40% for small dose, 60% for medium dose and 80% for high dose, while the improvement rate of hemorrhage in indirubin group was 30% for small dose, 50% for medium dose and 60% for high dose. There was no statistically significant difference in the improvement rate of hemorrhage between the two groups (P＞0.05). Compared with the model control group, PLT and Hb increased in different doses of SXXBC and indirubin group 4th-8th day after drug intervention (P＜0.05, 0.01). However, there was no significant difference between the different doses of SXXBC group and indirubin group (P＞0.05). Compared with the model control group, the WBC in each group was significantly lower (P＜0.05, 0.01) on the 4th-8th day after drug intervention; However, there was no statistical significance between the two groups of SXXBC and indirubin (P＞0.05). Compared with the model control group, the total number of megakaryocytes in each treatment group were decreased (P＜0.05, P＜0.01), in which the number of primary megakaryocytes in the large and medium dose groups of SXXBC and indirubin were decreased (P＜0.05, 0.01), and the number of juvenile megakaryocytes in the large dose group of SXXBC and indirubin were also decreased (P＜0.05). The number of granular megakaryocytes were decreased in each intervention groups (P＜0.05, 0.01), and the number of thromocytogenic megakaryocyte was increased in the high and medium dose groups of SXXBC and indirubin (P＜0.01). The time of prothrombin was shortened in the high and medium dose groups of SXXBC and indirubin (P＜0.05), and the fibrinogen (FIB) content in the high and medium dose groups of SXXBC was close to that of the normal control group. CONCLUSION: Both of the SXXBC and the indirubin standard all show good hemostatic effects. Indirubin shows a positive effect on increasing the peripheral platelet and hemoglobin in ITP model mice, regulating the immune response, reducing the total number of bone marrow megakaryocytes, increasing the thromocytogenic megakaryocyte, and increasing coagulation function.

Wholly Visible-Light-Responsive Host-Guest Supramolecular Gels Based on Methoxy Azobenzene and ß-Cyclodextrin Dimers.

Much attention has been paid to construct photoresponsive host-guest supramolecular gels; however, red-shifting the responsive wavelength remains a formidable challenge. Here, a wholly visible-light-responsive supramolecular gel was fabricated through the host-guest interaction between a ß-cyclodextrin (ß-CD) dimer and a tetra-ortho-methoxy-substituted azobenzene (mAzo) dimer (binary gelator) in DMSO/H2O (V/V = 8/2). The minimum gelation concentration of the low-molecular-weight binary gelator was 6 wt % measured via the tube inversion method. The substituted methoxy groups shifted the responsive wavelengths of trans-mAzo and cis-mAzo to the green and blue light regions, respectively. The host-guest interaction between mAzo and ß-CD as the driving force for gelation was confirmed using the 1H-NMR and 2D 1H NOESY spectra. The supramolecular gel showed good self-supporting ability with a storage modulus higher than 104 Pa. The release of Rhodamine B loaded in the gel as a model drug could be controlled by green light irradiation. We envisioned the potential applications of the wholly visible-light-responsive supramolecular compounds ranging from biomedical materials to smart materials.

Expression and clinical significance of WWOX, Elf5, Snail1 and EMT related factors in epithelial ovarian cancer.

Expression and clinical significance of WW domain-containing oxidoreductase (WWOX), Elf5, Snail1 and epithelial-mesenchymal transition (EMT) related factors in epithelial ovarian cancer were investigated. Ovarian cancer tissues of 300 epithelial ovarian cancer patients and the adjacent normal tissues were analyzed. Immunohistochemical method was used to detect the expressions of WWOX, Elf5, Snail1 and EMT marker molecules in the specimens. The relationship between the indicators and clinicopathological parameters, and prognosis of patients with ovarian cancer was analyzed. The relationship between WWOX, Elf5, Snail1 and EMT marker molecules E-cadherin, N-cadherin and vimentin in ovarian cancer tissues was analyzed. The expression levels of WWOX, Elf5, Snail1 and EMT marker molecules in epithelial ovarian cancer tissues were significantly different from those in adjacent normal tissues, and were related to surgical pathological stage, pathological grade and lymph node metastasis. High expressions of WWOX and Elf5 were related to the survival rate of patients. The survival rate of patients with positive expression was significantly higher than that of negative expression. FIGO stage, pathological grade, lymph node metastasis and expression of WWOX and Elf5 were all independent factors affecting postoperative prognosis in ovarian cancer patients. In conclusion, the expression levels of WWOX, Elf5, Snail1 and EMT related factors in epithelial ovarian cancer tissues are consistent and different. The expression levels of WWOX and Elf5 are related to the survival and prognosis of patients with epithelial ovarian cancer.

Reversible photo-responsive gel-sol transitions of robust organogels based on an azobenzene-containing main-chain liquid crystalline polymer.

Stimuli-responsive supramolecular gels have been widely investigated, but the construction of a liquid crystalline gel with a high mechanical property and reversible photo-response still remains a challenge. This is due to the difficulty of designing gelators with liquid crystal properties and gelation abilities in organic solvents simultaneously. In this study, an azobenzene-containing main-chain polyester (Azo-mLCP) with a pendant amide group was synthesized. The organogel of Azo-mLCP via a hydrogen bond in dioxane possessed reversible thermal- and photo-responsive behaviours. The organogel exhibited a good self-supporting ability when the concentration of the gelator was more than 7.5 wt%. The rapid trans-to-cis isomerization of Azo-mLCP in solution was studied via UV-Vis absorption spectra. In addition, the gel-to-sol transition of the organogel could be triggered efficiently by an incomplete trans-to-cis conversion strategy. This study opens a way for the main-chain liquid crystalline polymers to serve in potential applications in photo-responsive robust actuators, electro-optical devices, and so on.

LncRNA FAM83H-AS1 contributes to the radioresistance, proliferation, and metastasis in ovarian cancer through stabilizing HuR protein.

Ovarian cancer (OC) is a major cause of cancer-related deaths in women all over the world. The easy metastasis of OC and the problem of radioresistance are serious issues remaining to be overcome. Thus, research on molecular mechanisms underlying is in urgent demand. Long non-coding RNAs (lncRNAs) are a class of RNAs without protein coding potential, which has been reported to participate in the regulation on multiple biological process in cancers, including cell radiosensitivity and metastasis. Present study aimed to explore the role of lncRNA FAM83-AS1 in radioresistance and metastasis of ovarian cancer. First of all, the obvious upregulation of FAM83H-AS1 was identified by qRT-PCR in OC tissues, especially in metastatic tissues, as well as in OC cell lines. Importantly, we confirmed the correlation of FAM83H-AS1 levels with both ovarian cancer cells and normal ovarian cells. And Kaplan-Meier analysis indicated FAM83H-AS1 as a potential target of poor prognosis of OC. Through loss-of-function assays, we validated the inductive effect of FAM83H-AS1 in OC cell metastasis and radioresistance. Through mechanism research on FAM83H-AS1, we confirmed its interaction with HuR using pull-down assay and RNA immunoprecipitation (RIP), and verified the stabilization of HuR protein by FAM83-AS1 through western blot with the addition of CHX. Finally, rescue assays showed that overexpression of HuR rescued the suppression on radioresistance and metastasis in OC cell caused by the silencing of FAM83H-AS1. In conclusion, present study proved that FAM83H-AS1 contributes to the radioresistance and cell metastasis in ovarian cancer through stabilizing HuR protein.

Expression of ELF5 in endometrial carcinoma tissues and its clinical significance.

Expression of E74-like factor 5 (ELF5) in endometrial carcinoma tissues and its clinical significance were investigated. Eighty-four endometrial carcinoma tissues, 30 cases of atypical hyperplasia of endometrium and 30 cases of normal endometrial tissues were selected. Immunohistochemical method was utilized to detect the expression of ELF5 in different endometrial tissues. Moreover, its correlation with clinical pathological indexes of patients with endometrial carcinoma was analyzed. The postoperative follow-up was conducted in all the patients with endometrial carcinoma until June 30th, 2017. The Kaplan-Meier method was used for survival analysis so as to analyze the association of ELF5 expression level with clinical pathological indexes; Cox's proportional hazards regression model was utilized for univariate and multivariate analyses to screen independent risk factors for prognosis of endometrial carcinoma. In normal endometrial tissues, atypical hyperplasia and endometrial carcinoma tissues, the positive expression rates of ELF5 showed a decreased tendency (P=0.016). The positive expression rate of ELF5 in endometrial carcinoma tissues was lower in comparison to normal endometrial tissues (P=0.016). The expression of ELF5 was in accordance with the International Federation of Gynecology and Obstetrics (FIGO) staging of endometrial carcinoma (P<0.05), pathological grading (P<0.05), pathological typing (P=0.001), state of lymph node metastasis (P<0.05) and depth of myometrial invasion (P<0.05). Kaplan-Meier method for survival analysis showed that the average survival time of patients with negative ELF5 expression was shorter in comparison to the patients with positive expression (P=0.004). FIGO staging (P=0.004), pathological grading (P=0.048), depth of myometrial invasion (P=0.024) and lymph node metastasis (P=0.020) were related to the prognosis of patients with endometrial carcinoma, The univariate Cox's regression model analysis indicated that FIGO staging (P=0.010), pathological grading (P=0.040), depth of myometrial invasion (P=0.037), lymph node metastasis (P=0.029) and ELF5 (P=0.010) were associated with the prognosis of patients with endometrial carcinoma. Further, multivariate analysis revealed that ELF5 was an independent risk factor for prognosis of patients with endometrial carcinoma (P=0.035). The expression of ELF5 has a correlation with the occurrence, development and prognosis of endometrial carcinoma.

ELF5 in epithelial ovarian carcinoma tissues and biological behavior in ovarian carcinoma cells.

The expression of E74-like factor 5 (ELF5) in epithelial ovarian carcinoma tissues and its effects on biological behavior in ovarian carcinoma cells were assessed in search for a new approach for gene treatment of epithelial ovarian carcinoma. RT-PCR technology was applied to detect the expression of ELF5 mRNA in epithelial ovarian carcinoma (n=49), borderline ovarian epithelial tumor (n=19), benign ovarian epithelial tumor (n=31) and normal ovarian tissues (n=40). Then, we transfected recombinant plasmid pcDNA3.1­ELF5+EGFP into human ovarian carcinoma SKOV3 cells (recombinant plasmid group) in vitro and screened out stably transfected cells to conduct multiplication culture. Western blot analysis was performed to detect the expression of ELF5 protein in the different groups. Flow cytometry was employed to detect cell apoptosis and cycles. ELF5 mRNA in epithelial ovarian carcinoma and borderline ovarian epithelial tumor tissues were significantly lower (P<0.05) than those in benign ovarian epithelial tumor and normal ovarian tissues. ELF5 protein expression in the cells of recombinant plasmid group was significantly higher compared with empty plasmid and blank control groups. The capacity of cell reproductive recombinant plasmid group at each time point decreased (P<0.05). Flow cytometry detection showed that 67.03% of cells in recombinant plasmid group was blocked in G0/G1 phase (P<0.05), compared with empty plasmid group (37.17%) and blank control group (38.24%). Apoptotic rate of recombinant plasmid group was significantly lower (31.4±1.9%; P<0.05), compared with that of empty plasmid group (9.1±2.2%) and blank control group (8.7±1.5%), and the differences were statistically significant. In conclusion, ELF5 interfered with cell cycle of human ovarian carcinoma SKOV3 cells and promoted apoptosis of human ovarian carcinoma SKOV3 cells inhibiting their growth and invasive capacity; and thus providing a new approach to gene treatment of ovarian carcinoma.

Effect of the WWOX gene on the regulation of the cell cycle and apoptosis in human ovarian cancer stem cells.

In order to examine new ideas for gene therapy in ovarian cancer, the specific mechanism underlying the effects of the WW domain containing oxidoreductase (WWOX) gene on cell cycle regulation and apoptosis in human ovarian cancer stem cells was investigated. Ovarian cancer stem cells were transfected with a eukaryotic expression vector carrying the WWOX gene in vitro (recombinant plasmid) and cells transfected with the empty plasmid (empty plasmid) or untransfected cells were used as controls. Stably transfected cells were screened and amplified in culture and the WWOX protein was detected by western blot analysis in the three groups of cells. Western blot analysis was performed to detect the expression of cell cycle regulatory proteins cyclin E, cyclin-dependent kinase (CDK) 2, cyclin D1, CDK4 and apoptosis-related protein Wnt-5α and c-Jun N-terminal kinase (JNK), while polymerase chain reaction (PCR) was used to detect alterations in the mRNA expression levels of caspase-3. The results demonstrated that the WWOX protein was stably expressed in cells of the recombinant plasmid group, but was not detected in cells of the empty plasmid group and the control group. Cell proliferation at each time point decreased significantly in the recombinant plasmid group compared with the empty plasmid group and the control group. Flow cytometric analysis demonstrated that the proportion of cells in the G0/G1 phase in the recombinant plasmid group was significantly higher than that of cells in the empty plasmid group and the control group. The rate of apoptosis in the recombinant plasmid group was significantly higher than that of cells in the empty plasmid group and the control group. Western blot analysis demonstrated that the expression levels of cyclin E, CDK2, cyclin D1 and CDK4 in the recombinant plasmid group were significantly lower than those in the empty plasmid group and the control group; however, the expression levels of Wnt-5α and JNK were significantly higher than those in the empty plasmid group and the control group. PCR results demonstrated that the mRNA expression level of caspase-3 in the recombinant plasmid group was significantly higher than that in the empty plasmid group and the control group. In conclusion, the present study demonstrated that the WWOX gene can be stably expressed in ovarian cancer stem cells and that it inhibits the proliferation of ovarian cancer stem cells. The WWOX gene can downregulate the expression levels of cell cycle proteins cyclin E-CDK2 and cyclin D1-CDK4, which affects the cell cycle of ovarian cancer stem cells. Furthermore, the WWOX gene can upregulate the mRNA expression levels of Wnt-5α, JNK and caspase-3, thus contributing to apoptosis of ovarian cancer stem cells. The present study demonstrated that the WWOX gene may be an important molecular target for the treatment of ovarian cancer in the future.

Evaluation of the mechanism of epithelial-mesenchymal transition in human ovarian cancer stem cells transfected with a WW domain-containing oxidoreductase gene.

The aim of the present study was to investigate the impact of the WW domain-containing oxidoreductase (WWOX) gene on the mechanisms underlying epithelial-mesenchymal transition (EMT) in human ovarian cancer stem cells. Western blot analysis was performed to detect the differences in the expression of the EMT markers, E-cadherin, ß-catenin, N-cadherin, vimentin and fibronectin, between human ovarian cancer stem cells and the human epithelial ovarian carcinoma cell line, HO-8910. A pcDNA3.1-WWOX eukaryotic expression vector was subsequently transfected into the ovarian cancer stem cells (recombinant plasmid group) or an empty plasmid (empty plasmid group) and non-transfected ovarian cancer stem cells (blank control group) served as the controls. Following the transfection of the WWOX gene, methyl thiazolyl tetrazolium cell viability and Transwell® invasion assays, and western blot analysis were performed to detect changes in the proliferative capability and invasive capacity of ovarian cancer stem cells, as well as the expression of EMT markers and regulatory factors, Elf5 and Snail. The expression levels of E-cadherin and ß-catenin in the ovarian cancer stem cells were identified to be significantly lower than those in the HO-8910 cells, whereas the expression levels of N-cadherin, vimentin and fibronectin in the ovarian cancer stem cells were found to be significantly higher than those in the HO-8910 cells. At each time point, the cellular proliferative capacity of the recombinant plasmid group was observed to be significantly lower than that of the empty plasmid or blank control groups (P<0.05 vs. the controls). The number of penetrating cells in the recombinant plasmid, empty plasmid and the blank control groups were 105.5±3.1, 199.7±3.4 and 191.4±4.1, respectively (mean ± standard error of the mean; P<0.05 vs. the controls). In addition, the protein expression of E-cadherin, ß-catenin and Elf5 in the recombinant plasmid group was found to be significantly higher than that in the other two groups, whereas the protein expression of N-cadherin, vimentin, fibronectin and Snail in the recombinant plasmid group was significantly lower than that in the other two groups. An EMT exists in ovarian cancer stem cells, and the WWOX gene inhibits the cellular proliferation of ovarian cancer stem cells and reduces their invasive capability. Therefore, the WWOX gene may reverse the EMT in ovarian cancer stem cells by regulating the expression of the EMT regulatory factors, Elf5 and Snail.

Effects of 5-Aza-2'-deoxycytidine on the methylation state and function of the WWOX gene in the HO-8910 ovarian cancer cell line.

The aim of this study was to explore the effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR), a DNA methylation inhibitor, on the methylation state and function of the WWOX gene in the HO-8910 ovarian cancer cell line. The HO-8910 cells were divided into two groups, a control group and a 5-Aza-CdR-treated group. The methylation state of the WWOX gene was evaluated using a methylation-specific PCR assay. The effect of 5-Aza-CdR on the HO-8910 cells was analyzed using MTT and cell invasion assays, as well as flow cytometry. The animal models were established by intraperitoneal transplantation of the cells into nude mice. Following treatment with 5-Aza-CdR, a demethylation state was detected in the HO-8910 cells. WWOX protein expression was significantly higher in the 5-Aza-CdR-treated group compared with that in the control group. The cell growth rate at each tested time point and the number of invasive cells were lower in the 5-Aza-CdR-treated group compared with that in the control group. Flow cytometry revealed that 67.13% of the cells were arrested at the G0/G1 stage in the 5-Aza-CdR-treated group. The tumorigenic ability of the 5-Aza-CdR-treated group was lower compared with that of the control group. In conclusion, the methylation state of the WWOX gene in HO-8910 cells may be reversed using 5-Aza-CdR, which may also inhibit the growth of these cells.

Methylation status of WWOX gene promoter CpG islands in epithelial ovarian cancer and its clinical significance.

WW domain-containing oxidoreductase (WWOX) is a newly identified tumor suppressor gene that is associated with abnormal DNA methylation. The aim of this study was to evaluate the methylation status of CpG islands in the WWOX gene promoter region in cases of epithelial ovarian cancer and explore the correlation between the methylation status of the WWOX gene CpG islands and clinicopathological indices in patients with epithelial ovarian cancer. The methylation status of the WWOX gene CpG island was evaluated by methylation-specific polymerase chain reaction (MSP) in 48 patients with epithelial ovarian cancer, 18 patients with borderline epithelial ovarian tumors, 26 patients with epithelial benign tumors and 33 patients with normal ovarian tissues. Results showed that the rates of CpG island methylation in the WWOX gene promoter region in epithelial ovarian cancer tissues, borderline ovarian tumor tissues and benign ovarian tumor tissues were 43.75, 26.32 and 3.84%, respectively. The WWOX gene CpG islands were not methylated in normal ovarian tissues. The rate of CpG island methylation in epithelial ovarian cancer tissues was higher than that of other ovarian tissues and these differences were found to be statistically significant (P<0.01). The rate of CpG island methylation in the WWOX gene promoter region in late-stage (stage III and IV) epithelial ovarian cancer tissues was higher than that of early-stage (stage I and II) epithelial ovarian cancer tissues, and these differences were found to be statistically significant (P<0.05). In conclusion, epithelial ovarian cancer tissues showed CpG island hypermethylation in the WWOX gene promoter region, which may be an important mechanism leading to WWOX gene inactivation. Atypical methylation of WWOX gene is associated with the formation and progression of epithelial ovarian cancer, rendering it a potentially important indicator in the early diagnosis and prognosis of epithelial ovarian cancer.

Effects of sodium valproate on the growth of human ovarian cancer cell line HO8910.

To explore a possible new treatment for human ovarian cancer, we studied the effects of sodium valproate on the growth of the HO8910 human cell line. HO8910 cells were cultured in vitro and treated with different concentrations of sodium valproate. Cell proliferation, cell cycling, and apoptosis were measured by flow cytometry, cell morphology under a microscope, and expression levels of WWOX and P27 by Western blotting and RT-PCR. Tumor xenografts were established to determine in vivo effects of sodium valproate. Our results showed that cell proliferation was decreased with increasing concentration of sodium valproate, with features of cytoplasmic retraction and floating cells. Moreover, cell cycle analysis revealed a higher apoptosis rate and G0/ G1 phase in the sodium valproate experimental group than in the control group. In addition, protein expression levels of WWOX and P27 were elevated. Importantly, sodium valproate decreased in vivo xenograft tumor burden and up-regulated WWOX and P27 expression in nude mice. In conclusion, sodium valproate might play a role in inhibition and control of ovarian cancer cell line HO8910 by inhibiting cell proliferation, interfering with the cell cycle and promoting apoptosis, so that it may be effective in the clinical treatment of ovarian cancer.

[Effects of WWOX gene transfection on cell growth of epithelial ovarian cancer].

OBJECTIVE: To study the effects of anti-oncogene WWOX on cell growth of epithelial ovarian cancer,in order to find a new approach of gene therapy for ovarian cancer. METHODS: A eukaryotic expression vector containing WWOX was transfected into ovarian cancer cell line HO8910 in vitro (recombinant plasmid group), and positive cell clones were selected and amplified. Expression of WWOX protein was detected by western blot. Untransfected cell (blank contrast group) and transfected empty plasmid cell (empty plasmid group) were served as control groups. In vitro, the biology effect of WWOX on HO8910 cell was analyzed through the methyl thiazolyl tetrazolium test, transwell chamber cell invasion assay in vitro, agarose clony-formation and flow cytometry. In vivo, the cell of transfection was transplanted intraperitoneally in to BALB/c nude mice. The survival time and growth ability of nude mice were observed. RESULTS: (1) Recombinant plasmid group cell could steadily express WWOX protein, while in empty plasmid group and blank control group the expression of WWOX protein were not detected. (2) The growth rate of recombinant plasmid group cell was inhibited. (3) The agarose clony-formation rate of recombinant plasmid group (19.8%) was significantly lower than that of the empty plasmid group (54.5%) and blank control group (56.0%, P < 0.05). (4) Flow cytometry showed that (72.08 +/- 0. 39)% of cells was arrested at G0/G1 stage in recombinant plasmid group, while in empty plasmid group and blank control group G0/G1 stage cells were at (41.02 +/- 1.08)% and (39.31 +/- 0.67)% (P < 0.05). (5) In vitro invasion assay showed that invasion cell number in recombinant plasmid group (89.7 +/- 3.1) was not significantly different from that of empty plasmid group (91.2 +/- 1.3) and blank control group (91.4 +/- 1.3, P > 0.05). (6) In vivo test in nude mice showed that WWOX gene could inhibit tumor growth of the HO8910 cells. CONCLUSIONS: Tumor suppressor gene WWOX could interfere with the cell cycles of ovarian cancer cell and inhibit cell proliferation. As a new valuable tool, it promises to have application in the gene therapy of ovarian cancer.