CircRNA NALCN acts as an miR-493-3p sponge to regulate PTEN expression and inhibit glioma progression

Abstract Background An increasing number of studies have shown that circular RNAs (circRNAs) play important roles in the regulation of tumor progression. Therefore, we explored the expression characteristics, function, and related mechanism of the newly identified circNALCN in glioma. Methods RNA sequencing was used to analyze the expression profiles of circRNAs in brain tissue from five glioma cases and four normal controls. Quantitative real-time polymerase chain reaction was implemented to examine the levels of circNALCN, miR-493-3p, and phosphatase and tensin homolog (PTEN). Cell counting kit 8 assays were performed to analyze cell proliferation, and cell migration was assessed by the wound healing test and Transwell assay. Dual-luciferase reporter, fluorescence in situ hybridization, and RNA pulldown assays were performed to confirm the role of circNALCN as an miR-493-3p sponge, weakening the inhibitory effect of miR-493-3p on target PTEN expression. Results The downregulated expression of circNALCN was observed in both glioma tissues and cell lines. CircNALCN expression was negatively correlated with World Health Organization grade and overall survival in patients with glioma. Functionally, the overexpression of circNALCN significantly inhibited the proliferation and migration of glioma cells, whereas miR-493-3p mimics counteracted these effects. The mechanistic analysis demonstrated that circNALCN acted as a competing endogenous RNA for miR-493-3p to relieve the repressive effects of miR-493-3p on its target, PTEN, suppressing glioma tumorigenesis. Conclusions CircNALCN inhibits the progression of glioma through the miR-493-3p/PTEN axis, providing a developable biomarker and therapeutic target for glioma patients

Could extended laminectomy effectively prevent spinal cord injury due to spinal shortening after 3-column osteotomy?

Abstract Objective To explore whether the laminectomy extension can effectively prevent spinal cord injury (SCI) due to spinal shortening after 3-column osteotomy in goat models. Methods A total of twenty healthy goats were included and done with 3-column osteotomy of T13 and L1 under the somatosensory evoked potential (SSEP) monitoring. The samples were divided into two groups. The first group underwent regular laminectomy while the second group underwent an extended laminectomy with an extra 10 mm-lamina cranial to L2. The SSEP measured after 3-column osteotomy was set as the baseline, and the SSEP decreased by 50% from the baseline amplitude and/or delayed by 10% relative to the baseline peak latency was set as positive results, which indicated spinal cord injury. The vertebral column was gradually shortened until the SSEP monitoring just did not show a positive result. The height of the initial osteotomy gap (the distance from the lower endplate of T12 to the upper endplate of L2), the shortened distance (△H), the number of spinal cord angulated and the changed angle of the spinal cord (△α) were measured and recorded in each group. Neurological function was evaluated by the Tarlov scores on day 2 postoperatively. Results All the goats except one of the first group due to changes in the SSEP during the osteotomy were included and analyzed. In the first group, the height of the initial osteotomy segment and the safe shortening distances were 61.6 ± 2.6 mm and 35.2 ± 2.6 mm, respectively; the spinal cord of 5 goats was angulated (46.4 ± 6.6°), the other four goats were kinked and not angulated. In the second group, the height of the initial osteotomy segment and the safe shortening distances were 59.8 ± 1.5 mm and 43.3 ± 1.2 mm, respectively, and the spinal cord of ten goats were angulated (97.6 ± 7.2°). There was no significant difference in the height of the initial osteotomy segment between the two groups by using Independent-Samples T-Test, P = 0.095 (P > 0.05); there were significant difference in the safe shortening distance and the changed angle of the spinal cord between the two groups by using Independent-Samples T-Test (both $$\Delta$$ H and $$\Delta$$ α of P < 0.001), the difference between their mean were 8.1 mm and 51.2°. Significant difference was found in the number of spinal cord angulation between the two groups through Fisher’s exact test (5/9 vs. 10/10, P = 0.033). Conclusions An additional resection of 10 mm-lamina cranial to L2 showed the satisfactory effect in alleviating SCI after 3-column osteotomy. Timely and appropriate extend laminectomy could be a promising therapeutic strategy for SCI attributable to facilitating spinal cord angulation rather than spinal cord kinking and increasing the safe shortening distance

TRIM59 Promotes the Proliferation and Migration of Non-Small Cell Lung Cancer Cells by Upregulating Cell Cycle Related Proteins

<div><p>TRIM protein family is an evolutionarily conserved gene family implicated in a number of critical processes including inflammation, immunity, antiviral and cancer. In an effort to profile the expression patterns of TRIM superfamily in several non-small cell lung cancer (NSCLC) cell lines, we found that the expression of 10 TRIM genes including TRIM3, TRIM7, TRIM14, TRIM16, TRIM21, TRIM22, TRIM29, TRIM59, TRIM66 and TRIM70 was significantly upregulated in NSCLC cell lines compared with the normal human bronchial epithelial (HBE) cell line, whereas the expression of 7 other TRIM genes including TRIM4, TRIM9, TRIM36, TRIM46, TRIM54, TRIM67 and TRIM76 was significantly down-regulated in NSCLC cell lines compared with that in HBE cells. As TRIM59 has been reported to act as a proto-oncogene that affects both Ras and RB signal pathways in prostate cancer models, we here focused on the role of TRIM59 in the regulation of NSCLC cell proliferation and migration. We reported that TRIM59 protein was significantly increased in various NSCLC cell lines. SiRNA-induced knocking down of TRIM59 significantly inhibited the proliferation and migration of NSCLC cell lines by arresting cell cycle in G2 phase. Moreover, TRIM59 knocking down affected the expression of a number of cell cycle proteins including CDC25C and CDK1. Finally, we knocked down TRIM59 and found that p53 protein expression levels did not upregulate, so we proposed that TRIM59 may promote NSCLC cell growth through other pathways but not the p53 signaling pathway.</p></div

Knocking down of TRIM59 decreased the expression of cell cycle proteins.

<p>H1299 cells transiently transfected with TRIM59 siRNA-1, TRIM59 siRNA-2 or control siRNA were cultured in RPMI 1640 with 10% FBS for 48 hrs. <b>(A)</b> The mRNA expression levels of G2/M phase related genes CDC2, CCNA1, CCNB1, CCNB2 and CDC25C were tested by quantitative Real-Time PCR. <b>(B)</b> The protein expression levels of CDK1 (also known as CDC2), CDC25C and cyclin B1 were checked by western blot with indicated antibodies.</p

Multicomponent Solar Cells with High Fill Factors and Efficiencies Based on Non-Fullerene Acceptor Isomers

Multicomponent organic solar cells (OSCs), such as the ternary and quaternary OSCs, not only inherit the simplicity of binary OSCs but further promote light harvesting and power conversion efficiency (PCE). Here, we propose a new type of multicomponent solar cells with non-fullerene acceptor isomers. Specifically, we fabricate OSCs with the polymer donor J71 and a mixture of isomers, ITCF, as the acceptors. In comparison, the ternary OSC devices with J71 and two structurally similar (not isomeric) NFAs (IT-DM and IT-4F) are made as control. The morphology experiments reveal that the isomers-containing blend film demonstrates increased crystallinity, more ideal domain size, and a more favorable packing orientation compared with the IT-DM/IT-4F ternary blend. The favorable orientation is correlated with the balanced charge transport, increased exciton dissociation and decreased bimolecular recombination in the ITCF-isomer-based blend film, which contributes to the high fill factor (FF), and thus the high PCE. Additionally, to evaluate the generality of this method, we examine other acceptor isomers including IT-M, IXIC-2Cl and SY1, which show same trend as the ITCF isomers. These results demonstrate that using isomeric blends as the acceptor can be a promising approach to promote the performance of multicomponent non-fullerene OSCs

TRIM59 promotes the proliferation of NSCLC cells.

<p><b>(A)</b> Low serum assay. The indicated NSCLC cell lines transiently transfected with TRIM59 siRNA-1, TRIM59 siRNA-2, control siRNA or untransfected were cultured in RPMI 1640 medium supplemented with 1% FBS. At the indicated times, cells were trypsinized and counted. The data represent the average of three independent experiments (mean±SD) (Top figures). Immunoblot of TRIM59 to check the knockdown efficiency of TRIM59 siRNAs in the indicated cell lines (Bottom figures). <b>(B)</b> Saturation density assay. The indicated NSCLC cells transiently transfected with TRIM59 siRNA-1, TRIM59 siRNA-2, control siRNA or untransfected were cultured in RPMI 1640 supplemented with 10% FBS for 6 days, trypsinized and counted. The data represent the average of three independent experiments (mean±SD). <b>(C)</b> Colony formation assay. Five hundred H1299 cells transiently transfected with TRIM59 siRNA-1, TRIM59 siRNA-2, control siRNA or untransfected were seeded in 6-well plates in RPMI 1640 with 5% FBS. After two weeks, cells were fixed and stained with crystal violet. Representative wells were photographed and shown.</p

Knocking down of TRIM59 arrests NSCLC cell cycle in G2 phase.

<p>HBE and H1299 cells were transiently transfected with TRIM59 siRNA-1, TRIM59 siRNA-2, control siRNA or untransfected were cultured in RPMI 1640 with 10% FBS for 48 hrs. <b>(A)</b> Adherent cells were collected and cell cycle analysis was done by flow cytometry. The inserts showed the proportion of cells for each phase and are marked with different colors (pink: G0/G1 phase, green: S phase, and gray: G2/M phase). <b>(B)</b> The ratio of the cells in each phase was counted. <b>(C)</b> H1299 cells were fixed and stained with anti-PCNA antibody (red) and DAPI (blue). <b>(D)</b>The protein expression levels of PCNA in H1299 cells were checked by western blot.</p

TRIM59 is highly expressed in NSCLC cell lines.

<p><b>(A)</b> The mRNA expression levels of 60 TRIM family genes in four NSCLC cell lines (H1299, H292, SPC-A1 and A549) and normal human bronchial epithelial (HBE) cell line were determined by Q-PCR. The data were organized in a heat map by using MEV software. The relative expression levels of the genes were shown in the color scale of 0–4.0435486 in green-red-black color scheme. <b>(B)</b> The mRNA expression levels of TRIM genes including TRIM3, TRIM7, TRIM14, TRIM16, TRIM21, TRIM22, TRIM29, TRIM59, TRIM66 and TRIM70 were significantly upregulated in NSCLC cell lines compared with that in HBE cells. <b>(C)</b> Schematic depiction of TRIM59. TRIM59 contains a RING finger domain (RING), a B-box2 domain (B2), two coiled-coil domains (CC) and a transmembrane domain (TM). <b>(D)</b> The expression of TRIM59 in 14 kinds of normal tissues was checked by western blot using TRIM59 antibody. <b>(E)</b> The expression of TRIM59 protein in four NSCLC cell lines and HBE cell line. Lysates from the cell lines were subjected to immunoblot analysis with TRIM59 antibody.</p

TRIM59 promotes NSCLC cell growth not through p53 signaling pathway.

<p>Four types of indicated NSCLC cells and HBE cells transiently transfected with TRIM59 siRNA-1, TRIM59 siRNA-2, control siRNA or untransfected were cultured in RPMI 1640 with 10% FBS for 48 hrs. The protein expression levels of p53 were checked by western blot using anti-p53 and anti-TRIM59 antibodies.</p