WDR34 affects PI3K/Akt and Wnt/ß-catenin pathways to regulates malignant biological behaviors of glioma cells.

PURPOSE: Glioma is the most prevalent primary intracranial tumor globally. WDR34, a member of the WDR superfamily with five WD40 repeats, is involved in the pathogenesis of several tumors. However, the role of WDR34 in glioma progression is unknown. METHODS: The expression and prognostic significance of WDR34 in glioma patients were analyzed using GEPIA. WDR34 expression was detected by qRT-PCR. Western blot was employed to determine the expression of Ki67, proliferating cell nuclear antigen (PCNA), matrix metallopeptidase (MMP)2, MMP9, phosphatase and tensin homolog, protein kinase B (Akt), phosphorylated Akt, ß-catenin, and c-Myc. CCK-8, BrdU incorporation assay, Transwell invasion assay, flow cytometry analysis, and measurement of caspase-3 and caspase-9 activities were conducted to examine the effects of WDR34 knockdown on glioma cells. RESULTS: WDR34 was upregulated in glioma, which predicted a poor prognosis in glioma patients. WDR34 knockdown inhibited cell proliferation and reduced the expression of Ki67 and PCNA in glioma cells. WDR34 knockdown repressed the invasive ability of glioma cells by decreasing MMP-2 and MMP-9 expression. WDR34 knockdown increased the apoptotic rate and caspase-3 and caspase-9 activities in glioma cells. The PI3K/Akt and Wnt/ß-catenin pathways were inhibited after WDR34 knockdown in glioma cells. Moreover, overexpression of Akt or ß-catenin reversed the function of WDR34 knockdown on proliferation, invasion, and apoptosis. WDR34 knockdown reduced tumor growth in vivo. CONCLUSIONS: WDR34 knockdown inhibited malignant biological behaviors of glioma cells by inactivating the PI3K/Akt and Wnt/ß-catenin signaling cascades.

In situ growth of corrosion resistant Mg-Fe layered double hydroxide film on Q235 steel.

HYPOTHESIS: In situ grown layered double hydroxide (LDH) is commonly used one of the anticorrosion ways for metal materials; Due to the dense growth of LDH on the metal surface, its special layered structure can effectively delay the corrosion rate of metal. METHODS: In this study, we use a hydrothermal method to successfully grow Mg-Fe LDH film on steel substrates based on self-supplied Fe3+ ions. The films were characterized by X-ray diffraction, scanning electron microscopy, and X-ray energy dispersive spectrometry. The potential corrosion resistance of the obtained Mg-Fe LDH film was confirmed using electrochemical impedance spectroscopy and polarization curves. FINDINGS: After systematic adjustment and parameter optimization, it was found that Mg-Fe LDH film exhibited the best growth morphology and comprehensive performance with an initial pH value of 10, Mg2+/urea ratio of 1:4 and reaction time of 12 h. The SEM and electrochemical results further demonstrated that Mg-Fe LDH film play a good protection effect on carbon steel surface. This study provides an important reference for the processing of anticorrosion LDHs film.

Pseudogene ANXA2P2 knockdown shows tumor-suppressive function by inhibition of the PI3K/PKB pathway in glioblastoma cells.

The pseudogene annexin A2 pseudogene 2 (ANXA2P2) is highly expressed in glioblastoma (GBM). However, its role and mechanism involved in the progression of GBM remain poorly understood. ANXA2P2 messenger RNA expression was measured by quantitative reverse transcription-polymerase chain reaction. The protein levels were detected by Western blot. Cell viability was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase (LDH) release assays. Cell invasive ability was investigated by the transwell assay and by epithelial-mesenchymal transition (EMT). Cell apoptosis was examined by flow cytometry. The results showed that ANXA2P2 expression was increased in GBM tissues and cells. Silencing of ANXA2P2 inhibited the activation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB) pathway in GBM cells. Knockdown of ANXA2P2 decreased cell viability, promoted LDH release, suppressed cell invasive ability, and EMT, and induced cell apoptosis in GBM cells. The addition of the PI3K/PKB activator 740Y-P abrogated the effects of ANXA2P2 knockdown on cell viability, LDH release, invasive ability, and apoptosis. In conclusion, knockdown of ANXA2P2 inhibited cell viability and invasion but promoted the apoptotic rate by suppressing the PI3K/PKB pathway in GBM cells. ANXA2P2 may represent a new target for the treatment of GBM.

The nuclear transporter importin-11 regulates the Wnt/ß-catenin pathway and acts as a tumor promoter in glioma.

Karyopherins mediate the macromolecular transport between the cytoplasm and the nucleus and participate in cancer progression. However, the role and mechanism of importin-11 (IPO11), a member of the karyopherin family, in glioma progression remain undefined. Effects of IPO11 on glioma progression were detected using CCK-8, colony formation assay, flow cytometry analysis, caspase-3 activity assay, and Transwell invasion assay. Western blot analysis was used to detect the expression of active caspase-3, active caspase-7, active caspase-9, N-cadherin, Vimentin, E-cadherin, ß-catenin, and c-Myc. The activity of Wnt/ß-catenin pathway was evaluated by the T-cell factor/lymphoid enhancer factor (TCF/LEF) transcription factor reporter assay. Results showed that IPO11 knockdown inhibited proliferation and reduced colony number in glioma cells. IPO11 silencing promoted the apoptotic rate, increased expression levels of active caspase-3, caspase-7, and caspase-9, and enhanced caspase-3 activity. Moreover, IPO11 silencing inhibited glioma cell invasion by suppressing epithelial-to-mesenchymal transition (EMT). Mechanistically, IPO11 knockdown inactivated the Wnt/ß-catenin pathway. ß-Catenin overexpression abolished the effects of IPO11 silencing on the proliferation, apoptosis, and invasion in glioma cells. Furthermore, IPO11 silencing blocked the malignant phenotypes and repressed the Wnt/ß-catenin pathway in vivo. In conclusion, IPO11 knockdown suppressed the malignant phenotypes of glioma cells by inactivating the Wnt/ß-catenin pathway.

LncRNA SAMMSON Knockdown Inhibits the Malignancy of Glioblastoma Cells by Inactivation of the PI3K/Akt Pathway.

Dysregulated lncRNAs are proposed to be tightly associated with the progression of various tumors including glioblastoma (GBM). LncRNA Survival Associated Mitochondrial Melanoma-Specific Oncogenic Non-Coding RNA (SAMMSON) has been reported to be an oncogenic lncRNA in several tumors. Nevertheless, the specific role and molecular mechanism of SAMMSON in GBM progression remain unknown. Expression of SAMMSON in GBM tissues and cells was detected by qRT-PCR. CCK-8 and LDH release assays were applied to evaluate cellular viability. Invasion effect was assessed by Transwell invasion assay and western blot analysis of E-cadherin and N-cadherin expression. Apoptosis was detected using flow cytometry analysis and caspase-3 activity assay. The protein levels of phosphatidylinositol-3-kinase (PI3K), phosphorylated (p)-PI3K, protein kinase B (Akt) and p-Akt were estimated by western blot. We found that SAMMSON was highly expressed in GBM tissues and cells. SAMMSON knockdown suppressed cell viability and increased LDH release in GBM cells. Moreover, SAMMSON silencing impeded the invasive ability of GBM cells by regulating epithelial-to-mesenchymal transition (EMT). Furthermore, SAMMSON downregulation increased the apoptotic rate and caspase-3 activity in GBM cells. Additionally, it was demonstrated that the PI3K/Akt pathway was inhibited following SAMMSON silencing in GBM cells. Rescue assays revealed that activation of the PI3K/Akt pathway by 740Y-P abolished SAMMSON knockdown-induced viability reduction, invasion suppression and apoptosis in GBM cells. Taken together, lncRNA SAMMSON knockdown inhibited the malignancy of GBM cells by inactivation of the PI3K/Akt pathway.

miR-454-3p exerts tumor-suppressive functions by down-regulation of NFATc2 in glioblastoma.

Glioblastoma (GBM) is the most prevalent and malignant primary brain tumor. Numerous studies have demonstrated that aberrant microRNAs (miRNAs) expression is involved in various pathogenesis events in GBM. miR-454-3p has been found to be downregulated in GBM, however, the role and underlying mechanism has not been fully investigated. The expression levels of miR-454-3p in GBM clinical tissues and cultured cell lines were measured by qRT-PCR. To evaluate the role of miR-454-3p in GBM, GBM cells were transfected with miR-454-3p inhibitor (anti-miR-454-3p)/control inhibitor (anti-miR-NC) or miR-454-3p mimic/control mimic (miR-NC). Online prediction algorithm Targetscan was used to predict the target genes of miR-454-3p. The dual luciferase reporter gene assay was performed to confirm whether nuclear factor of activated T cells c2 (NFATc2) was a direct biological target of miR-454-3p. We found that miR-454-3p expression was significantly decreased in both GBM tissues and cultured cell lines. Overexpression of miR-454-3p by transfection with miR-454-3p mimic suppressed cell proliferation and induced apoptosis of GBM cells. Nuclear factor of activated T cells c2 (NFATc2) was predicted as a target gene of miR-454-3p. We found that NFATc2 expression was significantly increased in both GBM tissues and cultured cell lines. Besides, expression levels of miR-454-3p were negatively associated with NFATc2 expression in GBM tissues. NFATc2 knockdown in GBM cells obviously inhibited cell proliferation and elevated apoptosis. Luciferase reporter assay revealed that NFATc2 was a target gene of miR-454-3p. miR-454-3p overexpression suppressed the NFATc2 expression, while inhibition of miR-454-3p induced the NFATc2 expression. Suppression of NFATc2 blocked the effects of miR-454-3p inhibitor on cell proliferation and apoptosis in GBM cells. The results indicated that miR-454-3p executed the tumor suppressive activity via downregulating NFATc2 in GBM.

Intraoperative perfusion-weighted imaging in non-enhanced glioma surgery.

BACKGROUND: The objective of the present study was to observe and analyze the significance of perfusion-weighted imaging for guiding the operation implementation for non-enhanced glioma, and analyze the estimation of the histopathological grade of the non-enhanced glioma and the accuracy of the degree of malignancy degree before surgery. METHODS: Fifty-six patients diagnosed with non-enhanced glioma through conventional magnetic resonance scanning were selected. Before surgery, conventional magnetic resonance scanning and perfusion-weighted imaging were performed on all patients. The property classification was performed with the perfusion-weighted imaging parameters: cerebral blood volume (CBV) and cerebral blood flow (CBF) before surgery. RESULTS: Surgery was performed on the 56 patients. Tumors were excised and processed for histopathological classification and semi-quantitative immunohistochemical analysis of vascular endothelial growth factor (VEGF) levels. Histology was compared after surgery and the classification accuracy rate was analyzed before surgery. Additionally, conventional magnetic resonance scanning and perfusion-weighted imaging were performed on 15 patients during surgery. We compared and analyzed the reference value of perfusion-weighted imaging before and during surgery. Residual diseased tissues were excised; histopathological examination was performed, and semi-quantitative immunohistochemical analysis of VEGF was performed. Regarding maximal magnetic resonance perfusion-weighted imaging measured before surgery, the CBV, CBF, and expression level of VEGF were positively correlated with the pathological grade of tumors. If the CBV and CBF values of the non-enhanced glioma were higher, the grade of malignancy was higher (P<0.01), and the positive expression rate of VEGF was higher (P<0.01). CONCLUSIONS: Magnetic resonance perfusion-weighted imaging can display vessel growth and distribution within non-enhanced gliomas before surgery, and effectively evaluate the histopathological grade and grade of malignancy, and provide accurate guidance for tumor resection during surgery.

MicroRNA-485-5p attenuates cell proliferation in glioma by directly targeting paired box 3.

MicroRNA-485-5p (miR-485-5p) has been reported to be involved in the development and progression of human cancers; however, its role in glioma remains unclear. In the present study, we found that miR-485-5p was significantly down-regulated in both glioma tissues and cell lines. Functional experiments indicated that enhanced expression of miR-485-5p attenuated glioma cell proliferation in vitro and in vivo, and induced glioma cells cycle arrest in G1. MiR-485-5p was found to directly bind to the 3'-UTR of paired box 3 (PAX3) and decrease its expression of protein level, which further inhibits the proliferation of glioma. The decreasing of PAX3 was found to lead to the accumulation of p-JNK. Mechanistic studies revealed that restoring the expression of PAX3 alleviated miR-485-5p-induced inhibition of proliferation of glioma cells. Taken together, these findings suggest that PAX3 modulation by miR-485-5p has an important role in regulating glioma proliferation, and miR-485-5p might be a novel therapeutic target for glioma.

Prognostic and clinical implication of a disintegrin and metalloprotease 8 expression in pediatric medulloblastoma.

AIM: To investigate the expression patterns and clinical value of a disintegrin and metalloprotease 8 (ADAM8) in pediatric medulloblastoma. METHODS: We evaluated the expression of ADAM8 mRNA and protein in pediatric medulloblastoma tissues by Quantitative RT-PCR, Western blot analysis and Immunohistochemical staining, respectively. The correlation of ADAM8 immunostaining with clinical-pathological features of medulloblastoma patients and its prognostic relevance were further determined. RESULTS: ADAM8 mRNA and protein were both most highly expressed in medulloblastoma tissues when compared with normal cerebellum tissues (both P<0.001). According to the immunohistochemisty analysis, we found statistically significant correlations of high ADAM8 protein expression with advanced metastatic stage (P=0.01), aggressive histopathological type (P=0.006), as well as with undifferentiated tumor (P=0.02). We further determined a statistically significant association between ADAM8 protein expression and clinical outcome of medulloblastoma patients, with higher expression levels of ADAM8 associating with a worse overall survival (P=0.02). CONCLUSION: Our results provide the first evidence that the up-regulation of ADAM8 in medulloblastoma tissues might be correlated with the advanced tumor progression and poor prognosis of patients with this disease. Detection of ADAM8 expression might facilitate the prognostic assessment and improve the therapeutic strategy for medulloblastoma patients.

Synthesis and application in polypropylene of a novel of phosphorus-containing intumescent flame retardant.

A novel phosphorus-containing triazine oligomer poly(2-morpholinyl-4-penta-erythritol phosphate-1,3,5-triazine) (PMPT) was synthesized as a kind of tri-component intumescent flame retardant (IFR). The chemical structure of PMPT was characterized by FTIR, 1H-NMR and 31P-NMR, and the mechanical and flammability properties of FR-PP were measured. The FTIR results showed that the expected chemical reactions had  happened at each step. The 1H-NMR and 31P-NMR spectra also agreed with the chemical structure of PMPT. The slight effect of PMPT on the mechanical properties of FR-PP suggested that PMPT and PP are compatible. The high limited oxygen index (LOI) values of FR-PP revealed that PMPT was an efficient IFR and there was the synergistic effect between PMPT and ammonium polyphosphate/ pentaerythritol (APP/PER).