METTL14 inhibits Aß1-42-induced neuronal injury through regulating the stability of CBLN4 mRNA in Alzheimer's disease.

Previous studies have suggested that N6-methyladenosine (mA) modification of RNA affects fundamental aspects of RNA metabolism, and mA dysregulation is implicated in various human diseases, including Alzheimer's disease (AD). This study is designed to explore the role and mechanism of methyltransferase-like 14 (METTL14) in the pathogenesis of AD. SK-N-SH cells were treated with Aß1-42 to establish an in vitro model of AD. Cerebellin 4 (CBLN4) and METTL14 expression levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability and apoptosis were analyzed using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and flow cytometry assay. B-cell lymphoma-2 (Bcl-2), Bcl-2 related X protein (Bax), C-caspase-3, total-caspase-3, C/EBP homologous protein (CHOP), and glucose-related protein 78 (GRP78) protein levels were determined using Western blot. Interleukin-1ß (IL-1ß) and tumor necrosis factor α (TNF-α) levels were analyzed using ELISA. Reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) products were examined using special assay kits. Interaction between CBLN4 and METTL14 was verified using methylated RNA immunoprecipitation (MeRIP) and dual-luciferase reporter assays. CBLN4 and METTL14 expression was decreased in Aß1-42-treated SK-N-SH cells. Upregulation of CBLN4 relieved Aß1-42-induced SK-N-SH cell apoptosis, inflammation, oxidative stress, and endoplasmic reticulum (ER) stress in vitro. At the molecular level, METTL14 could improve the stability and expression of CBLN4 mRNA via m6A methylation. Our findings indicated that m6A methylase METTL14-mediated upregulation of CBLN4 mRNA stability could repress Aß1-42-triggered SK-N-SH cell injury, providing a promising therapeutic target for AD treatment.

PVT1 regulates hippocampal neuron apoptosis and inflammation in epilepsy by miR-206-3p-dependent regulation of CAMK4.

This study was designed to dissect the function of plasmacytoma variant translocation 1 (PVT1) in hippocampal neuron injury in epilepsy and its possible molecular basis. Status epilepticus (SE) mouse model was built and primary hippocampal neurons were isolated. qRT-PCR and Western blot were applied to quantify the levels of related genes and proteins. Cell proliferation and apoptosis were examined by CCK-8, EdU, and flow cytometry assays. Inflammatory factors were detected using ELISA analysis. Dual-luciferase reporter and RIP assays were carried out to validate the relationship between miR-206-3p and PVT1 or CAMK4. PVT1 and CAMK4 were increased, and miR-206-3p was downregulated in the hippocampus and hippocampal neurons of SE mice. Knockdown of PVT1 or CAMK4 abated SE-induced proliferation inhibition, apoptosis, and inflammation in hippocampal neurons. Mechanistically, PVT1 could sponge miR-206-3p to upregulate the expression of CAMK4 in hippocampal neurons. Moreover, downregulation of miR-206-3p reversed the inhibitory effects of PVT1 knockdown on SE-induced apoptosis and inflammation in hippocampal neurons. Similarly, overexpression of CAMK4 abolished miR-206-3p-evoked arrest of apoptosis and inflammation in hippocampal neurons under SE condition. Collectively, PVT1 contributed to SE-induced apoptosis and inflammation in hippocampal neurons by modulating the miR-206-3p/CAMK4 axis, offering a novel insight into the prevention of epilepsy.

CENPA facilitates glioma stem cell stemness and suppress ferroptosis to accelerate glioblastoma multiforme progression by promoting GBP2 transcription.

The function of glioma stem cells (GSCs) is closely related to the progression of glioblastoma multiforme (GBM). Centromere protein A (CENPA) has been confirmed to be related to the poor prognosis of GBM patients. However, whether CENPA regulates GSCs function to mediate GBM progression is still unclear. GSCs were isolated from GBM cells. The expression of CENPA and guanylate-binding protein 2 (GBP2) was examined by quantitative real-time PCR and western blot. GSCs proliferation and stemness were assessed using EdU assay and sphere formation assay. Cell ferroptosis was evaluated by detecting related factors. The interaction between CENPA and GBP2 was analyzed by ChIP assay and dual-luciferase reporter assay. Animal experiments were conducted to measure the effect of CENPA knockdown on the tumorigenicity of GSCs in vivo. CENPA was upregulated in GBM tissues and GSCs. CENPA knockdown inhibited GSCs proliferation, stemnness, and promoted ferroptosis. GBP2 was overexpressed in GBM tissues and GSCs, and CENPA enhanced GBP2 transcription by binding to its promoter region. CENPA overexpression accelerated GSCs proliferation and stemnness and suppressed ferroptosis, while GBP2 knockdown reversed these effects. Downregulation of CENPA reduced the tumorigenicity of GSCs by decreasing GBP2 expression in vivo. In conclusion, CENPA enhanced GBP2 transcription to increase its expression, thus accelerating GSCs proliferation and stemnness and repressing ferroptosis. Our findings promote a new idea for GBM treatment.

Inhibition of the TCF12/VSIG4 axis by palbociclib diminishes the proliferation and migration of glioma cells and decreases the M2 polarization of glioma-associated microglia.

The CDK4/CDK6 inhibitor palbociclib has shown the encouraging promise in the treatment of glioma. Here, we elucidated how palbociclib exerts suppressive functions in the M2 polarization of glioma-related microglia and the progression of glioma. Xenograft experiments were used to evaluate the function in vivo. The mRNA levels of transcription factor 12 (TCF12) and VSIG4 were detected by RT-qPCR, and their protein levels were assessed by immunoblotting. Cell migration was tested by wound-healing assay. Cell cycle distribution and M1/M2 microglia phenotype analysis were performed by flow cytometry. The levels of IFN-γ, TNF-α, IL-6，and TGF-ß were measured by ELISA. The TCF12/VSIG4 association was verified by luciferase reporter and chromatin immunoprecipitation (ChIP) assays. In U251 and LN229 glioma cells, TCF12 and VSIG4 were overexpressed, and palbociclib reduced their expression levels. TCF12 upregulation enhanced the proliferation and migration of glioma cells and the M2 polarization of glioma-associated microglia in vitro as well as the tumorigenicity of U251 glioma cells in vivo, which could be reversed by palbociclib. Mechanistically, TCF12 could enhance VSIG4 transcription and expression by binding to the VSIG4 promoter. TCF12 deficiency led to repression in glioma cell proliferation and migration as well as microglia M2 polarization, which could be abolished by increased VSIG4 expression. Our study reveals the novel TCF12/VSIG4 axis responsible for the efficacy of palbociclib in combating glioma, offering a rationale for the application of palbociclib in glioma treatment.

USP9X deubiquitinates TRRAP to promote glioblastoma cell proliferation and migration and M2 macrophage polarization.

Glioblastoma (GBM) is the most aggressive form of brain cancer, characterized by rapid growth and invasion into surrounding brain tissue. Ubiquitin-specific protease 9X (USP9X) has emerged as a key regulator in various cancers, but its role in GBM pathogenesis remains unclear. Understanding the molecular mechanisms underlying USP9X modulation of GBM progression could unveil potential therapeutic targets for this deadly disease. The mRNA and protein levels were determined in GBM tissues and/or cells using quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting assays, respectively. Cell migration was evaluated through wound-healing assay, while cell proliferation was measured using colony formation and CCK-8 assays. Flow cytometry analysis was performed to quantify the CD206-positive macrophages to assess M2 polarization. Co-immunoprecipitation (Co-IP) assays were conducted to elucidate the association between USP9X and transformation/transcription domain-associated protein (TRRAP). Cycloheximide (CHX) treatment was used to determine the impact of USP9X on TRRAP protein stabilization. Furthermore, the effect of USP9X depletion on GBM cell malignancy was validated using a xenograft mouse model. We found that USP9X expression was elevated in GBM tissues and cells. Depletion of USP9X suppressed GBM cell migration, proliferation, and M2 macrophage polarization. Mechanistically, USP9X stabilized TRRAP through the deubiquitination pathway in GBM cells, and TRRAP mitigated the effects of USP9X silencing on GBM cell malignant phenotypes and M2 macrophage polarization. Moreover, silencing of USP9X inhibited tumor formation in vivo. Together, USP9X deubiquitinated TRRAP, thereby promoting glioblastoma cell proliferation, migration, and M2 macrophage polarization. These results highlight the potential of targeting the USP9X-TRRAP axis as a therapeutic strategy for GBM.

SKA3 promotes glioblastoma proliferation and invasion by enhancing the activation of Wnt/ß-catenin signaling via modulation of the Akt/GSK-3ß axis.

Spindle and kinetochore-related complex subunit 3 (SKA3) is a key modulator of the progression of multiple tumor types. However, the involvement of SKA3 in glioblastoma (GBM) has not been well studied. The current study aimed to explore the role of SKA3 expression and the potential function of the protein in GBM. Our data showed that SKA3 expression was significantly up-regulated in GBM. Functional assays demonstrated that the knockdown of SKA3 impeded the proliferation, colony formation and invasion of GBM cells, while SKA3 overexpression produced the opposite effects. Further investigation revealed that SKA3 overexpression enhanced the activation of Wnt/ß-catenin signaling, which was associated with the enhanced phosphorylation of Akt and glycogen synthase kinase-3ß (GSK-3ß). Notably, the inhibition of Akt markedly abrogated the SKA3 overexpression-induced promotion of Wnt/ß-catenin signaling in GBM cells. Further, the inhibition of Wnt/ß-catenin signaling markedly abrogated the SKA3 overexpression-induced promotion of tumor growth. In addition, the knockdown of SKA3 significantly retarded tumor formation and GBM progression in vivo. In summary, these data demonstrate that SKA3 exerts promotes tumor growth in GBM by enhancing the activation of Wnt/ß-catenin signaling via modulation of the Akt/GSK-3ß axis. This work highlights the pivotal role of SKA3/Akt/GSK-3ß/Wnt/ß-catenin signaling in the progression of GBM and suggests that SKA3 is an attractive therapeutic target with potential to be used to treat GBM.

LHPP exerts a tumor-inhibiting role in glioblastoma via the downregulation of Akt and Wnt/ß-catenin signaling.

Phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPP) has been recently identified as a novel inhibitor of multiple tumors; however, its role in glioblastoma (GBM) has not been investigated. This study aimed to evaluate whether LHPP exerts a potential tumor-inhibiting role in GBM. Compared with that in normal tissues, LHPP expression was lower in GBM tissues and various GBM cell lines. LHPP up-regulation in GBM cells markedly reduced their proliferation and invasion, and its knockdown had an oncogenic effect on these cells. Further studies revealed that overexpressed LHPP decreased the levels of Akt and glycogen synthase-3ß phosphorylation and down-regulated Wnt/ß-catenin signaling. By contrast, LHPP knockdown produced opposite effects. Akt suppression markedly abrogated the activation of Wnt/ß-catenin signaling induced by LHPP knockdown. The reactivation of Wnt/ß-catenin signaling partially reversed the inhibition of tumor growth in GBM mediated by LHPP overexpression. In addition, LHPP overexpression markedly retarded the tumorigenesis of GBM cells in vivo. These findings revealed that LHPP acts a potential inhibitor of tumor growth in GBM, and its overexpression represses GBM proliferation and invasion by down-regulating Akt and Wnt/ß-catenin signaling. This work highlights the crucial role of LHPP in GBM progression and suggests its potential as an anticancer target for the treatment of this disease.

A Framework of Combining Short-Term Spatial/Frequency Feature Extraction and Long-Term IndRNN for Activity Recognition.

Smartphone-sensors-based human activity recognition is attracting increasing interest due to the popularization of smartphones. It is a difficult long-range temporal recognition problem, especially with large intraclass distances such as carrying smartphones at different locations and small interclass distances such as taking a train or subway. To address this problem, we propose a new framework of combining short-term spatial/frequency feature extraction and a long-term independently recurrent neural network (IndRNN) for activity recognition. Considering the periodic characteristics of the sensor data, short-term temporal features are first extracted in the spatial and frequency domains. Then, the IndRNN, which can capture long-term patterns, is used to further obtain the long-term features for classification. Given the large differences when the smartphone is carried at different locations, a group-based location recognition is first developed to pinpoint the location of the smartphone. The Sussex-Huawei Locomotion (SHL) dataset from the SHL Challenge is used for evaluation. An earlier version of the proposed method won the second place award in the SHL Challenge 2020 (first place if not considering the multiple models fusion approach). The proposed method is further improved in this paper and achieves 80.72% accuracy, better than the existing methods using a single model.

Single Image De-Raining via Improved Generative Adversarial Nets.

Capturing images under rainy days degrades image visual quality and affects analysis tasks, such as object detection and classification. Therefore, image de-raining has attracted a lot of attention in recent years. In this paper, an improved generative adversarial network for single image de-raining is proposed. According to the principles of divide-and-conquer, we divide an image de-raining task into rain locating, rain removing, and detail refining sub-tasks. A multi-stream DenseNet, termed as Rain Estimation Network, is proposed to estimate the rain location map. A Generative Adversarial Network is proposed to remove the rain streaks. A Refinement Network is proposed to refine the details. These three models accomplish rain locating, rain removing, and detail refining sub-tasks, respectively. Experiments on two synthetic datasets and real world images demonstrate that the proposed method outperforms state-of-the-art de-raining studies in both objective and subjective measurements.

Efficacy of percutaneous vertebroplasty treatment of spinal tumors: A meta-analysis.

OBJECTIVE: To evaluate the function of percutaneous vertebroplasty (PVP) treatment to pain relief and life quality for patients with spinal tumors. METHODS: Articles about the researches on the treatment of spinal tumors by PVP in PubMed, Embase, and the Chinese Biomedical Literature database from January 1, 2015 to December 31, 2013. The keywords "spinal tumors," "efficacy," and "vertebroplasty" were firstly scanned to exclude all irrelevant articles. Then, the final inclusion of studies was determined by reading the full text of the remaining articles. The citation lists of all retrieved articles were scanned to identify other potentially relevant reports. RevMan5.2 was used to analyze pain intensity visual analog scale (VAS) and Karnofsky performance scores (KPS) within each research. Combined HRs (hazard ratio) were calculated using fixed- or random- effects models according to the heterogeneity. RESULTS: Twenty-six studies involving 1351 patients met our selection criteria. Meta-analysis results among 10 case-control studies showed that the combined HR was -2.83 [95% confidence interval (CI) -2.92, -2.73; P < .0001], indicating a 2.83-fold decrease of pain in PVP group. For 12 single-arm studies, a significantly decrease of pain after PVP treatment (HR = -4.79, 95% CI -5.00, -4.57, P < .0001) was also found in PVP group. In addition, for KPS analysis, the combined HR was 16.31 (95% CI 14.31, 18.31; P < .0001), which indicated that PVP treatment was associated with a 16.31-fold increase of KPS. The combined HR was 0.58 (95% CI 0.35, 0.96; P = .04) for complication analysis. CONCLUSIONS: PVP treatment of spinal tumor is significantly associated with better pain relief and life quality, which could improve the outcome in metastatic spinal tumor patients.

CDC7-dependent transcriptional regulation of RAD54L is essential for tumorigenicity and radio-resistance of glioblastoma.

Accumulating evidence indicates that cell division cycle 7-related protein kinase(CDC7) plays an essential role in tumor cells and it could induces cell proliferation and could be related to prognosis in multiple types of cancer. However, the biological role and molecular mechanism of CDC7 in GBM still remains unclear. In this study, we identified that CDC7 expression was enriched in glioblastoma (GBM) tumors and was functionally required for tumor proliferation and its expression was associated to poor prognosis in GBM patients. Mechanically, CDC7 induced radio resistance in GBM cells and CDC7 knock down increased cell apoptosis when combined with radiotherapy. Moreover, CDC7 regulated The DNA repair/recombination protein 54L (RAD54L) expression via regulation of RAD54L promoter activity. Therapeutically, we found that CDC7 inhibitor attenuated tumor growth both in vitro and in vivo. Collectively, CDC7 promotes proliferation, induces radio resistance in GBM, and could become a potential therapeutic target for GBM.

MicroRNA-613 impedes the proliferation and invasion of glioma cells by targeting cyclin-dependent kinase 14.

Increasing evidence has suggested that microRNAs (miRNAs) are critical regulators of tumorigenesis. MicroRNA-613 (miR-613) has recently been reported as a novel tumor-related miRNA that plays an important role in multiple cancers. However, the expression and functional significance of miR-613 in glioma remains unclear. In this study, we aimed to investigate the biological function of miR-613 in glioma. We found that miR-613 expression was frequently downregulated in glioma tissues and cell lines compared with normal controls. Overexpression of miR-613 impeded proliferation and colony formation and induced cell cycle arrest in G0/G1 phase, and also inhibited the invasive ability of glioma cells. By contrast, miR-613 inhibition had the opposite effects. Bioinformatic analysis and dual-luciferase reporter assays showed that miR-613 directly targets the 3'-untranslated region of cyclin-dependent kinase 14 (CDK14). Real-time quantitative PCR and Western blot analysis showed that CDK14 expression is negatively regulated by miR-613. In addition, miR-613 expression was inversely correlated with CDK14 expression in clinical glioma tissues. Moreover, overexpression of miR-613 decreased the protein expression of ß-catenin and inhibited the activation of Wnt signaling. Importantly, the antitumor effects of miR-613 were significantly reversed by CDK14 overexpression. Overall, our results show that miR-613 inhibits glioma cell proliferation and invasion by downregulating CDK14, suggesting that miR-613 and CDK14 may serve as potential therapeutic targets for the treatment of glioma.

Tetrandrine protects against oxygen-glucose-serum deprivation/reoxygenation-induced injury via PI3K/AKT/NF-κB signaling pathway in rat spinal cord astrocytes.

Tetrandrine (TET) is a bis-benzylisoquinoline alkaloid, which is isolated from a Chinese medicinal herb with antioxidant and anti-inflammatory activities. In this study, we investigated the effects of TET on oxygen-glucose-serum deprivation/reoxygenation (OGSD/R)-induced injury in rat spinal cord astrocytes, which mimics hypoxic/ischemic conditions in vivo. MTT and LDH assays indicated that cell viability was distinctly reduced and LDH leakage was elevated after OGSD/R exposure, which were dose-dependently reversed by pretreatment with TET (0.1, 1, 10, 20µM). Western blot analysis showed that OGSD/R exposure resulted in an enhanced expression of Bax and Caspase-3 proteins, and Bcl-2 reduction; whereas these effects were dose-dependently restored by TET pretreatment. TET pretreatment also dose-dependently inhibited the elevated Caspase-3 activity in OGSD/R-treated astrocytes. The oxidative stress status was evaluated using commercial kits, and the results demonstrated that OGSD/R exposure induced obvious oxidative stress, accompanied by elevated levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and reduced superoxide dismutase (SOD) activity, which were dose-dependently restored by TET pretreatment. In addition, TET pretreatment diminished the accumulation of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) induced by OGSD/R. Moreover, TET pretreatment dose-dependently suppressed Akt phosphorylation and nuclear factor-kappaB (NF-κB) activity augmented by OGSD/R. Similarly, both PI3K inhibitor LY294002 and NF-κB inhibitor PDTC notably attenuated OGSD/R-induced Akt phosphorylation, NF-κB activation, ROS generation, and TNF-α secretion. Taken together, these data demonstrated that TET protected against OGSD/R-induced injury in rat spinal cord astrocytes, which may be attributed to its antioxidant and anti-inflammatory activities via PI3K/AKT/NF-κB signaling pathway.

Carvacrol attenuates traumatic neuronal injury through store-operated Ca(2+) entry-independent regulation of intracellular Ca(2+) homeostasis.

Searching for effective pharmacological agents for traumatic brain injury (TBI) treatment has largely been unsuccessful. The transient receptor potential melastatin 7 (TRPM7), a TRP channel that is essential for embryonic development, has been shown to mediate ischemic neuronal injury in vivo and in vitro, but global deletion of TRPM7 in mice is lethal. Here, carvacrol was used to investigate the protective effect of TRPM7 inhibition in an in vitro traumatic neuronal injury model. Carvacrol (0.5 and 1 mM) reduced lactate dehydrogenase (LDH) release, apoptosis and caspase-3 activation after traumatic injury in cortical neurons. These neuroprotective effects were accompanied by alleviated cytoplasmic calcium levels as measured by calcium imaging. In contrast, the thapsigargin (TG) induced store-operated calcium entry (SOCE) and the expression of SOCE related proteins in neurons were not altered by carvacrol treatment. The involvement of TRPM7 sensitive calcium influx in our in vitro model was confirmed by the results that bradykinin induced calcium influx was prevented by carvacrol in neurons. Furthermore, carvacrol significantly inhibited the induction of neuronal nitric oxide synthase (nNOS) after traumatic injury, and treatment with carvacrol and the nNOS inhibitor NLPA together had no extra effect on calcium concentration and neuronal injury. Thus, inhibition of TRPM7 function by carvacrol protects against traumatic neuronal injury, and might be a potential drug development strategy for the treatment of TBI.

[Characterization of glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and its association with cell proliferation and apoptosis].

OBJECTIVE: To investigate the glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and the association between cell proliferation and apoptosis and the metabolic status. METHODS: An in vitro hypoxic cell model was established in SHG44 cells using CoCl2. Real-time PCR and Western blotting were used to assess the expressions of hypoxia-inducible factor-1α (HIF-1α) and the enzymes involved in glycolysis including PDK1, PKM2, and LDHA. Intracellular ATP levels were measured by bioluminescence assay to assess the energy metabolic status of SHG44 cells. The viability and apoptosis of the cells were examined using MTT assay and flow cytometry, respectively. RESULTS: The cells in hypoxic culture showed obviously increased expressions of HIF-1α, LDHA, PDK1, and PKM2 at both the mRNA and protein levels as compared to those in normal cell culture. Hypoxia of the cells also resulted in a lowered cell proliferative activity and an increased apoptosis rate with lowered intracellular ATP concentrations and elevated mitochondrial membrane potential. CONCLUSION: Hypoxia can induce a glycolytic phenotype of tumor cells. The sensitivity of tumor cells to hypoxia-induced cell death is directly correlated with their metabolic status.