LncRNA CCAT2 promotes angiogenesis in glioma through activation of VEGFA signalling by sponging miR-424.

Glioma is characterized by high morbidity, high mortality and poor prognosis. Recent studies exhibited that lncRNA CCAT2 is overexpressed in glioma and promotes glioma progression, but the specific molecular biological mechanism remains to be determined. We performed qRT-PCR to evaluate the expression of related genes, Western blotting analysis to measure protein levels, colony formation assay to detect the proliferative ability of glioma cells, flow cytometry to measure cell apoptosis, bioinformatics analysis and dual luciferase assay to verify the binding sites and the targeted regulatory relationship in A172 and U251 cell lines and tube formation assay to determine endothelial angiogenesis. LncRNA CCAT2 and VEGFA were highly expressed, while miR-424 was expressed at low levels in NHA cells. Furthermore, knockdown of lncRNA CCAT2 decreased cell proliferation, increased cell apoptosis and inhibited endothelial angiogenesis in glioma. Moreover, lncRNA CCAT2 shared a complementary sequence with miR-424 which in turn directly bound to the 3'-UTR of VEGFA. Further investigation indicated that lncRNA CCAT2 promoted cell proliferation and endothelial angiogenesis by inducing the PI3K/AKT signalling pathway in glioma. The oncogenic lncRNA CCAT2 is highly associated with the development of glioma and exerts its function by upregulating VEGFA via miR-424.

A Novel Peptide from Abalone (Haliotis discus hannai) to Suppress Metastasis and Vasculogenic Mimicry of Tumor Cells and Enhance Anti-Tumor Effect In Vitro.

Vasculogenic mimicry (VM) formed by tumor cells plays a vital role in the progress of tumor, because it provides nutrition for tumor cells and takes away the metabolites. Therefore, the inhibition of VM is crucial to the clinical treatment of tumors. In this study, we investigated the anti-tumor effect of a novel peptide, KVEPQDPSEW (AATP), isolated from abalone (Haliotis discus hannai) on HT1080 cells by migration, invasion analysis and the mode of action. The results showed that AATP effectively inhibited MMPs by blocking MAPKs and NF-κB pathways, leading to the downregulation of metastasis of tumor cells. Moreover, AATP significantly inhibited VM and pro-angiogenic factors, including VEGF and MMPs by suppression of AKT/mTOR signaling. In addition, molecular docking was used to study the interaction of AATP and HIF-1α, and the results showed that AATP was combined with an active site of HIF-1α by a hydrogen bond. The effect of AATP on anti-metastatic and anti-vascular in HT1080 cells revealed that AATP may be a potential lead compound for treatment of tumors in the future.

LncRNA PART1 inhibits glioma proliferation and migration via miR-374b/SALL1 axis.

BACKGROUND: Abnormal expression of lncRNA is involved in a diversity of diseases and plays a vital role in targeted therapy. However, few studies have been conducted on lncRNA PART1 in glioma. We aimed to investigate the function and the potential regulatory mechanism of lncRNA PART1/miR-374b/SALL1 axis in glioma. METHODS: qRT-PCR and western blotting detected genes and proteins expression. Dual-luciferase reporter assay was performed to examine the binding relationship of lncRNA PART1, miR-374b, and SALL1. MTT assay and clone formation assay were performed to detect the cell viability and proliferation. Transwell assay detected glioma cell migration. In vivo tumor development experiments detected changes in tumor size, volume, and weight of the tumor after overexpression of lncRNA PART1. Immunohistochemistry was used to detect ki-67, E-cadherin, and N-cadherin expression. RESULTS: The expression of lncRNA PART1 and SALL1 were down-regulated and miR-374b was up-regulated in different glioma cell lines. Overexpression of lncRNA PART1 inhibited glioma cell proliferation, migration, and epithelial mesenchymal transition (EMT). LncRNA PART1 targeted miR-374b to promote SALL1 expression. The knockdown of miR-374b inhibited glioma cell proliferation and migration and EMT by SALL1. What's more, overexpression of miR-374b or knockdown of SALL1 reversed the inhibitory effect of lncRNA PART1 on the proliferation, migration, and EMT of glioma cells. Furthermore, overexpression of lncRNA PART1 inhibited glioma growth in vivo. CONCLUSION: LncRNA PART1 inhibited glioma proliferation and migration via miR-374b/SALL1 axis. These results might provide new insights for comprehending the complex lncRNA-miRNA network in gliomas.

miR-376a inhibits glioma proliferation and angiogenesis by regulating YAP1/VEGF signalling via targeting of SIRT1.

BACKGROUND: Glioma is the most common cancer in the central nervous system. Previous studies have revealed that the miR-376 family is crucial in tumour development; however, its detailed mechanism in glioma is not clear. METHODS: Cellular mRNA or protein levels of miR-376a, SIRT1, VEGF and YAP1 were detected via qRT-PCR or Western blotting. We analysed the proliferation, angiogenesis and migration abilities of glioma cell lines using colony formation, tube formation and Transwell assays. A luciferase assay was performed to determine whether miR-376a could recognize SIRT1 mRNA. Xenograft experiments were performed to analyse the tumorigenesis capacity of glioma cell lines in nude mice. The angiogenesis marker CD31 in xenograft tumours was detected via immunohistochemistry (IHC). RESULTS: miR-376a expression was lower in glioma cells than in normal astrocytes. miR-376a mimic inhibited SIRT1, YAP1, and VEGF expression and suppressed the proliferation, migration and angiogenesis abilities of the glioma cell lines LN229 and A172, whereas miR-376a inhibitor exerted the opposite functions. In a luciferase assay, miR-376a inhibited the luciferase activity of WT-SIRT1. SIRT1 overexpression upregulated YAP1 and VEGF in glioma cells and promoted proliferation, migration and angiogenesis. Xenografts with ectopic miR-376a expression exhibited lower volumes and weights and a slower growth curve. Overexpression of miR-376a inhibited YAP1/VEGF signalling and angiogenesis by inhibiting SIRT1 in xenograft tissues. CONCLUSION: miR-376a directly targets and inhibits SIRT1 in glioma cells. Downregulation of SIRT1 resulted in decreased YAP1 and VEGF signalling, which led to suppression of glioma cell proliferation, migration and angiogenesis.

miR-503 Inhibits Proliferation, Migration, And Angiogenesis Of Glioma By Acting On VEGFA Through Targeting LRIG2.

BACKGROUND: Glioma is a common malignant tumor of the human central nervous system, and the pathological characteristics include invasive growth, angiogenesis, and so on. Ectopic expression of miR-503 works as a critical factor in cancer cell proliferation, migration, and capillary-like tube formation. The potential mechanisms of miR-503 in angiogenesis of glioma cells are still not reported. METHODS: The expression levels of miR-503, LRIG2, and VEGFA mRNA and protein were performed by quantitative reverse transcription-PCR or Western blot assay. Dual-Luciferase reporter gene assay was used to determine the interaction between miR-503 and LRIG2. The concentration of VEGFA was measured using the ELISA method. The cell proliferation, migration, and angiogenesis of cocultured HCMEC/D3 cells were analyzed by MTT assay, transwell detection, and tube formation assay, respectively. RESULTS: The expression levels of LRIG2 and VEGFA were reduced in glioma cells with miR-503 overexpression and enhanced with miR-503 inhibition. Moreover, cell proliferation, migration, and angiogenesis of cocultured HCMEC/D3 cells were alleviated with miR-503 mimics transfection. VEGFA and miR-503 inhibitor promoted cell proliferation, cell migration, and angiogenesis. Luciferase reporter gene assay revealed that miR-503 could directly target LRIG2. Furthermore, knockdown of LRIG2 or addition of VEGF inhibitor bevacizumab could abrogate the effect of miR-503 inhibitor on VEGFA expression, as well as the promotion of cell proliferation, migration, and angiogenesis. CONCLUSION: MiR-503 mediated LRIG2 suppression and regulated the expression of VEGFA, thereby reducing cell proliferation, migration, and angiogenesis of glioma cells. These results provide new insight into the action mechanism of miR-503-modulated signaling pathway in angiogenesis of glioma cells.

Diaqua-bis(benzyl-oxyacetato)copper(II).

In the title mononuclear complex, [Cu(C(9)H(9)O(3))(2)(H(2)O)(2)], the Cu(II) ion, located on an inversion center, is hexa-coordinated by four O atoms from two benzyl-oxyacetate ligands [Cu-O = 1.9420 (14) and 2.2922 (14) Å] and two water mol-ecules [Cu-O = 2.0157 (15) Å] in a distorted octa-hedral geometry. In the crystal structure, inter-molecular O-H⋯O hydrogen bonds link the mol-ecules into layers parallel to the bc plane.

Diaqua-bis[2-(benzyl-oxy)acetato]cobalt(II).

In the mononuclear title complex, [Co(C(9)H(9)O(3))(2)(H(2)O)(2)], each Co(II) atom is located on an inversion center and is hexa-coordinated by four O atoms from two benzyl-oxyacetate ligands [Co-O bond lengths = 2.0487 (9) and 2.1090 (9) Å] and two water mol-ecules [Co-O bond length = 2.0873 (9) Å] in a distorted octa-hedral geometry. In the crystal structure, inter-molecular hydrogen bonds and π-π stacking inter-actions [centroid-centroid distance between phenyl rings = 3.692 (2) Å] link the mol-ecules into a supra-molecular structure.

[Behavior and Degradation of Polycyclic Aromatic Hydrocarbons (PAHs) in Coking Wastewater of A/O2 and A/O/H/O Processes].

Polycyclic aromatic hydrocarbons (PAHs) are typical organic pollutants found in coking wastewater, and their behavior and reduction can be affected by different treatment processes. Based on these considerations, this study investigated the behaviors of PAHs in coking wastewater under A/O2 and A/O/H/O treatment processes, respectively. In order to evaluate variations in PAH removal under two different treatment processes, samples were taken from different treatment units for quantification of PAHs using gas chromatography-mass spectrometry. Results showed that PAHs were barely degraded in anaerobic tanks of either treatment process and accumulated much higher concentrations than in aerobic and hydrolytic tanks. While low molecular weight PAHs (LMW PAHs) in aqueous phase from anaerobic tanks were degraded effectively in aerobic tanks, high molecular weight PAHs (HMW PAHs) mostly accumulated in the sludge phase; these potentially pose a higher environmental risk and therefore need to be treated separately. Moreover, the A/O/H/O process showed higher degradation of PAHs bioavailability and higher removal effectiveness for PAHs with four or more benzene rings than the A/O2 process; this is attributed to the hydrolytic tank's ability to promote hydrolysis of macromolecular organic compounds and therefore improve biodegradability of PAHs. Comprehensive results from the study indicated that the A/O/H/O process is more advantageous for degradation of PAHs than the A/O2 process.

[Aerobic Degradation and Microbial Community Succession of Coking Wastewater with Municipal Sludge].

Coking wastewater is a typical industrial wastewater with high toxicity. Its treatment with biological processes is often challenging because it contains constituents inhibiting microbial activity. To study the inhibitory effect and possible acclimation of microbes in coking wastewater treatment, municipal sludge was inoculated into coking wastewater. Time-dependent concentrations of COD, phenol, ammonia nitrogen, and thiocyanide in coking wastewater were analyzed. The microbial community structure was investigated by the Illumina high-throughput sequencing technology during inoculation. The results showed that COD began to decrease after 16 h and 97.1% of phenol disappeared after 40 h. Thiocyanide began to degrade at 72 h and was undetectable after 96 h. Accordingly, the concentration of ammonia increased as the thiocyanide concentrations decreased. High-throughput pyrosequencing analysis showed that the microbial community structure and species richness varied at different culture stages. In the stage of phenol degradation, the abundance of Acinetobacter and Pseudomonas increased rapidly; the species richness was 13.04% of the community at 48 h. In the stage of thiocyanate degradation, Sphingobacterium,Brevundimonas,Lysobacter, and Chryseobacterium were the dominant bacteria and were 16.13% of the community at 96 h. At 144 h, Fluviicola,Stenotrophomonas, and Thiobacillus became the dominant species and were 22.45% of the community abundance. The results showed that municipal sludge can rapidly overcome the toxicity of coking wastewater because the pollutants are degraded rapidly. The microbial community structure changed as wastewater components were degraded. Environmental factors and the competition among bacteria played a key role in microbial community succession.

[Characteristics of Pahs pollution in sediments from Leizhou coastal marine area, Liusha Bay and Shenzhen Bay].

Leizhou coastal marine area, Liusha Bay and Shenzhen Bay represented open coastal area and half-closed bay, respectively. This study discussed the differences of PAHs concentration levels, spatial distribution and sources in sediments from these three marine areas. The results showed that detected ratios of 15 PAHs were 100%, and major compounds were 3-ring and 4-ring PAHs, especialy Phe, Fla, Pry and Bbf; Sigma PAHs concentration was Leizhou < Shenzhen < Liusha. In spatial distribution, PAHs concentrations were the east < the south < the west in Leizhou; the inside > the outside, and the aquaculture > the non-aquaculture in Liusha Bay and Shenzhen Bay. It suggested that large-scale mariculture inside bay played an important role in PAHs pollution and might make it serious. Oil, fossil fuels and biomass burning were the dominant sources of PAHs in sediments from Leizhou coastal area, Liusha Bay and Shenzhen Bay.

Effects of NAT2 polymorphism on SASP pharmacokinetics in Chinese population.

BACKGROUND: Sulfasalazine (SASP) pharmacologic actions are widely applied in clinical therapy. The role of N-Acetyltransferase 2 (NAT2) in the pharmacokinetics of SASP and its metabolites has not been clarified. We investigated the effects of genetic polymorphism of NAT2 on pharmacokinetic profiles of SASP and its two metabolites, sulfapyridine (SP) and N-acetylsufapyridine (AcSP). METHODS: Eighteen subjects were recruited and divided into 3 groups by NAT2 genotype: wild type (w/w), heterozygous variant (w/m), homozygous variant (m/m). After taking 1000mg SASP tablets, the plasma concentrations of SASP, SP and AcSP were measured with HPLC method and pharmacokinetic parameters were calculated by using the computing program 3P97. RESULTS: The AUC(0)(-)(72) and Cmax of SP in m/m subjects were significantly higher than those in w/m and w/w subjects, with the values of 172.57+/-49.42, 103.38+/-39.85, 71.37+/-17.52mg h/l, and 9.65+/-2.34, 6.10+/-1.79, 4.55+/-1.38mg/l, respectively. In contrast, the AUC(0)(-)(72) of AcSP was significantly lower in m/m subjects. The Cmax of AcSP in w/w, w/m and m/m subjects was 12.67+/-3.32, 9.07+/-2.29 and 4.22+/-0.93mg/l, respectively, with significant differences among groups. However, there was no significant difference in any pharmacokinetic parameter of SASP among groups. CONCLUSION: Different NAT2 genotypes, leading to functional heterogeneity of NAT2, may affect pharmacokinetics of SP and AcSP. Therefore, genotyping NAT2 gene before administration would be important in SASP therapy.

Modulation of signal transducers and activators of transcription (STAT) factor pathways during focal cerebral ischaemia: a gene expression array study in rat hippocampus after middle cerebral artery occlusion.

1. Signal transducers and activators of transcription (STAT) factors are a family of transcription factors that mediate intracellular signalling initiated at cytokine cell surface receptors and transmitted to the nucleus. In the present study, we determined the global changes in STAT gene expression in the hippocampus of rats after focal cerebral ischaemia and reperfusion using microarray analysis. 2. The present study used middle cerebral artery occlusion (MCAO) to induce ischaemia and reperfusion in Sprague-Dawley rats. Using superarray Q series Janus tyrosine kinases (Jak)/STAT signalling pathway gene array, a total of 96 genes was screened in adult male rat hippocampus after transient focal cerebral ischaemia. 3. The results showed that 23 genes were upregulated at least twofold by ischaemia treatment and that 12 genes were downregulated at least threefold by ischaemia treatment compared with controls. 4. After confirmation by quantitative real-time polymerase chain reaction, the data suggest that the gene expression of STAT2, 5a, 5b, 6 and suppressor of cytokine signalling (SOCS) 4 was increased by ischaemia, probably due to a compensatory response of the brain, which may play a protective role in damaged brain tissue. 5. The results of the present study provide evidence on global changes in STAT gene expression in the hippocampus of rats after focal cerebral ischaemia and reperfusion, in which STAT2, 5a, 5b, 6 and SOCS4 were confirmed to be significantly modulated during focal cerebral ischaemia.