Role In Glioma Cells Research Articles

MicroRNA-608 Regulates Epithelial-To-Mesenchymal Transition to Inhibit Malignant Behaviors of Glioma Cells Through Nuclear Factor I-C Signaling Pathway

MiR-608 is expressed in bladder cancer, colon cancer, prostate cancer risk, and exerts inhibitory role in gastric cancer and invasion of three negative malignant breast cancer by inhibiting the NIFC, but its role in the biology of glioma is unclear, so the purpose of this study was to explore MiR-608’s role in glioma, and further explore whether NFIC signal pathways are involved in the role of MiR-608 in glioma. We obtain glioma tissues and normal tissues to detect the expression of miR-608 by PCR. miR-608 mimics transfection or joint with NFIC expression plasmid were transfected into A172, and LN229 glioma cells followed by analysis of cell proliferation, clone formation, invasion and migration, and cell apoptosis. Compared to normal tissue, miR-608 expression in glioma tissues was decreased. After transfection of miR-608 mimics, miR-608 level, cell proliferation activity, invasion and migration activity increased significantly, and apoptosis reduced, in addition, the dual luciferase report gene and protein imprinting analysis showed NFIC to be a target of miR-608. NFIC transfection reversed miR-608’s role in glioma cells. In conclusion, microRNA-608 inhibits malignant invasion and migration of glioma cells via NFIC signaling.

Hypoxia-inducible factor 1α (HIF-1α)-activated Gli1 induces invasion and EMT by H3K4 methylation in glioma cells

Abstract Objectives Gliomas are highly aggressive neuroepithelial-layer malignancies. Hypoxia-inducible factor 1α (HIF-1α) was revealed to be upregulated in gliomas under hypoxia. Nevertheless, its role in glioma cells remains elusive. We attempted to clarify the molecular mechanism of HIF-1 underlying glioma. Methods Cellular models were established to mimic the characteristics of hypoxia. RT‒qPCR was used to detect HIF-1α and Gli1 levels in glioma cells with or without hypoxic treatment. Transwell assays were used to measure the invasive ability of U87 and U251 cells. Western blotting was used to evaluate epithelial-mesenchymal transition (EMT)-associated protein abundance and H3K4 methylation (H3K4me)-associated protein abundance in U87 and U251 cells. ChIP assessed the association of HIF-1α or H3K4me with the Gli1 promoter in glioma cells. Results HIF-1α and Gli1 were upregulated in glioma cells relative to normal human astrocytes (NHAs). HIF-1α and Gli1 were also upregulated in hypoxia-treated glioma cells relative to untreated glioma cells. Both HIF-1α and Gli1 silencing suppressed glioma invasion and EMT under hypoxia. HIF-1α upregulated Gli1 transcriptionally via MLL1-mediated H3K4me. H3K4me mutation silencing was further demonstrated to suppress glioma cell invasion and EMT under hypoxia. Conclusions Both HIF-1α and Gli1 are upregulated in glioma cells and function as oncogenes in glioma cells. HIF-1α transcriptionally activates Gli1 via MLL1-mediated H3K4 methylation in glioma cells, providing ideas for seeking new therapeutic directions for glioma.

LINC01094 Affects Glioma Cell Proliferation Through miR-204-3p

This study intends to assess LINC01094′s role in glioma cells. LINC01094 level in glioma tissue was detected by RT qPCR. After transfection of LINC01094 overexpression plasmid, LINC01094 siRNA, and si-LINC01094 and miR-204-3p inhibitor, cell proliferation was evaluated by MTT and cell invasion and migration was assessed by transwell and scratch test. LINC01094 expression in glioma tissues was significantly increased. Overexpression of LINC01094 can significantly promote cell proliferation, which was significantly inhibited after knockdown of LINC01094. In addition, silence of LINC01094 can upregulate miR-204-3p and inhibit cell proliferation and promote apoptosis induced by overexpression of LINC01094. In conclusion, LINC01094 promotes glioma cell proliferation through miR-204-3p.

A Metabolic Plasticity-Based Signature for Molecular Classification and Prognosis of Lower-Grade Glioma

Background: Glioma is one of the major health problems worldwide. Biomarkers for predicting the prognosis of Glioma are still needed. Methods: The transcriptome data and clinic information on Glioma were obtained from the CGGA, TCGA, GDC, and GEO databases. The immune infiltration status in the clusters was compared. The genes with differential expression were identified, and a prognostic model was developed. Several assays were used to detect RPH3A’s role in Glioma cells, including CCK-8, colony formation, wound healing, and transwell migration assay. Results: Lower Grade Glioma (LGG) was divided into two clusters. The immune infiltration difference was observed between the two clusters. We screened for genes that differed between the two groups. WGCNA was used to construct a co-expressed network using the DEGs, and four co-expressed modules were identified, which are blue, green, grey, and yellow modules. High-risk patients have a lower overall survival rate than low-risk patients. In addition, the risk score is associated with histological subtypes. Finally, the role of RPH3A was detected. The overexpression of RPH3A in LGG cells can significantly inhibit cell proliferation and migration and regulate EMT-regulated proteins. Conclusion: Our study developed a metabolic-related model for the prognosis of Glioma cells. RPH3A is a potential therapeutic target for Glioma.

Silencing long noncoding RNA LINC01138 inhibits aerobic glycolysis to reduce glioma cell proliferation by regulating the microRNA‑375/SP1 axis.

Glioma is a primary cerebral neoplasm that originates from glial tissue and spreads to the central nervous system. Long noncoding RNAs are known to play a role in glioma cells by regulating cell proliferation, migration and invasion. The aim of the present study was to investigate the mechanism by which long intergenic non-protein coding RNA (LINC) 01138 affects glycolysis and proliferation in glioma cells via the microRNA (miR)-375/specificity protein 1 (SP1) axis. LINC01138 expression was assessed in glioma tissues and cells using reverse transcription-quantitative PCR and the association between LINC01138 and patient clinicopathological features was analyzed. Glucose uptake, lactic acid secretion, cell proliferation, and glycolysis-related enzyme levels were detected following LINC01138 silencing using CCK-8, EDU assay and western blot analysis. miR-375 and SP1 expression levels were also assessed, and the distribution of LINC01138 in the nucleus and cytoplasm was investigated using subcellular fractionation localization. Furthermore, the binding relationships between LINC01138 and miR-375, and between miR-375 and SP1 were assessed via dual-luciferase experiment, RIP and RNA pull-down assays. Finally, xenograft transplantation models were used to verify the in vitro results. LINC01138 was highly expressed in glioma, which was independent of patient sex or age but was significantly related to tumor diameter, the World Health Organization tumor grade and lymph node metastasis. Silencing LINC01138 significantly reduced glioma glycolysis and cell proliferation. Moreover, LINC01138 acted as a competing endogenous RNA to sponge miR-375 and promote SP1 expression. miR-375 inhibition significantly reversed the effect of LINC01138 silencing. In addition, silencing LINC01138 significantly reduced tumor growth in vivo. The present study demonstrated that silencing LINC01138 inhibited aerobic glycolysis and thus reduced glioma cell proliferation, potentially by modulating the miR-375/SP1 axis.

LINC02381 contributes to cell proliferation and hinders cell apoptosis in glioma by transcriptionally enhancing CBX5

Glioma, featured with high incidence and low survival rate, is the most common type of primary brain tumor, severely affecting human life worldwide. LINC02381 is an interesting lncRNA functioning as oncogenic lncRNA in some cancers but as tumor-suppressor in others, but no report demonstrates its association with and function in glioma. Intriguingly, we found in a bioinformatics website LncRNADisease that LINC02381 was closely related to malignant glioma, so this study aimed to figure out the expression and function of LINC02381 in glioma. By RT-qPCR, we confirmed LINC02381 upregulation in glioma cells. Functional experiments demonstrated that LINC02381 knockdown repressed glioma cell proliferation and induced apoptosis. Boinformatics tools and RT-qPCR revealed the positive correlation between LINC02381 and CBX5 in glioma cells. More importantly, we confirmed that LINC02381 could interact and work synergistically with CEBPβ to bind to CBX5 promoter and activate CBX5 transcriptionally. Additionally, rescue experiments indicated that CBX5 up-regulation reversed the decline in cell proliferation and the augment in cell apoptosis caused by LINC02381 knockdown. To conclude, LINC02381 could facilitate CBX5 transcription via interaction with CEBPβ, thus exerting its oncogenic role in glioma cells, which could contribute to better understanding of glioma.

PUMA mediated afatinib-induced apoptosis in glioma cells

IntroductionAfatinib exhibits a tumor-inhibiting effect in different cancers by inducing apoptosis; however, its pro-apoptosis role in glioma cells is still not fully understood.Material and methodsIn the current study, two glioma cell lines (U373-MG and U87-MG) were treated with afatinib to measure their tolerance for afatinib-triggered cell death and apoptosis.ResultsWe found that afatinib treatment repressed both growth and proliferation and induced apoptosis in glioma cells. Moreover, it increased the expression of pro-apoptotic p53-upregulated modulator of apoptosis (PUMA). The influence of PUMA on afatinib-triggered apoptosis was assessed by PUMA overexpression and knockdown in glioma cell lines. PUMA overexpression resulted in an increased sensitivity of glioma cell lines toward afatinib, whereas its knockdown abated the effect of afatinib on apoptosis. Similarly, the in vivo potency of afatinib on U373-MG xenograft tumors in wild type (WT) and PUMA knockdown nude mice was measured. Afatinib treatment reduced the weight and volume of WT xenograft tumors but did not have the same effect on PUMA knockdown xenograft tumors. Afatinib also induced significant cell death and apoptosis in WT xenograft tumors but not in PUMA knockdown xenograft tumors.ConclusionsIn conclusion, afatinib induces apoptosis in glioma cells by mediating PUMA expression. This study warrants further investigation into the mechanism of afatinib in glioblastoma treatment.

Zinc Finger E-Box Binding Homeobox 1 Regulates the Biological Behavior of Glioma Cells via iNOS/NF-κB Signaling

The gliomas pathogenesis is complex and effective molecular targets are still unclear. ZEB1 regulates epithelial mesenchymal transition (EMT) and participates in tumors. Our study intends to analyze ZEB1’s role in glioma cells. qRT-PCR detected ZEB1 mRNA expression in normal group and tumor group. ZEB1 siRNA was transfected into glioma cells followed by measuring ZEB1, E-cadherin and Vimentin expression, cell proliferation, Capase-3 activity as well as NF-κB and iNOS changes by immunoblotting. Upregulation of ZEB1 was found in glioma tumor tissue and correlated with glioma clinicopathological characteristics. Interfering with ZEB1 by siRNA significantly down-regulated ZEB1, inhibited cell proliferation, increased Capase-3 activity, down regulated NF-κB and iNOS proteins in glioma cells, elevated E-cadherin and decreased Vimentin level (P <0.05). ZEB1 down regulation in glioma cells can change the expression of NF-κB/iNOS, regulate cell apoptosis and inhibit cell proliferation, thereby delaying EMT process.

Tanshinone IIA Suppresses Glioma Cell Proliferation, Migration and Invasion Both in vitro and in vivo Partially Through miR-16-5p/Talin-1 (TLN1) Axis.

BackgroundTanshinone IIA (TIIA) is one of the active constituents derived from the rhizome of Danshen, a traditional Chinese herbal. Recently, microRNAs (miRNAs) have been suggested to be associated with the anticancer role of TIIA. However, it remains vague of the interaction between miRNAs and TIIA in glioma, a common aggressive brain tumor in humans.MethodsExpression of miRNA (miR)-16-5p and talin-1 (TLN1) was detected using reverse transcription-quantitative polymerase chain reaction and Western blotting. Cell proliferation, migration and invasion were assessed with cell viability assay, transwell assay, Western blotting, and xenograft tumor experiment. The target binding between miR-16-5p and TLN1 was confirmed by dual-luciferase reporter assay and RNA pull-down assay.ResultsTIIA treatment inhibited cell viability, migration and invasion, and decreased Cyclin D1, matrix metalloproteinase (MMP)-9 and Vimentin expression in glioma T98G and A172 cells both in vitro and in vivo. Thus, TIIA induced anti-glioma role, wherein miR-16-5p was upregulated and TLN1 was downregulated. Moreover, silencing miR-16-5p could abate TIIA-mediated suppression on glioma cell proliferation, migration and invasion in vitro and in vivo. TLN1 overexpression also exerted tumor-promoting effect in TIIA-treated T98G and A172 cells. Mechanically, miR-16-5p could regulate TLN1 expression via target binding, and depleting TLN1 could counteract the inhibitory effect of miR-16-5p knockdown on the curative effect of TIIA in T98G and A172 cells.ConclusionTIIA exerted the anti-proliferation, anti-migration and anti-invasion role in glioma cells both in vitro and in vivo partially through regulating miR-16-5p/TLN1 axis.

Multifractal Properties of BK Channel Currents in Human Glioblastoma Cells.

Potassium channels play an important physiological role in glioma cells. In particular, voltage- and Ca2+-activated large-conductance BK channels (gBK in gliomas) are involved in the intensive growth and extensive migrating behavior of the mentioned tumor cells; thus, they may be considered as a drug target for the therapeutic treatment of glioblastoma. To enable appropriate drug design, molecular mechanisms of gBK channel activation by diverse stimuli should be unraveled as well as the way that the specific conformational states of the channel relate to its functional properties (conducting/nonconducting). There is an open debate about the actual mechanism of BK channel gating, including the question of how the channel proteins undergo a range of conformational transitions when they flicker between nonconducting (functionally closed) and conducting (open) states. The details of channel conformational diffusion ought to have its representation in the properties of the experimental signal that describes the ion-channel activity. Nonlinear methods of analysis of experimental nonstationary series can be useful for observing the changes in the number of channel substates available from geometrical and energetic points of view at given external conditions. In this work, we analyze whether the multifractal properties of the activity of glioblastoma BK channels depend on membrane potential, and which states, conducting or nonconducting, affect the total signal to a larger extent. With this aim, we carried out patch-clamp experiments at different levels of membrane hyper- and depolarization. The obtained time series of single channel currents were analyzed using the multifractal detrended fluctuation analysis (MFDFA) method in a standard form and incorporating focus-based multifractal (FMF) formalism. Thus, we show the applicability of a modified MFDFA technique in the analysis of an experimental patch-clamp time series. The obtained results suggest that membrane potential strongly affects the conformational space of the gBK channel proteins and the considered process has nonlinear multifractal characteristics. These properties are the inherent features of the analyzed signals due to the fact that the main tendencies vanish after shuffling the data.

Bit1 Silencing Enhances the Proliferation, Migration, and Invasion of Glioma Cells Through Activation of the IL-6/STAT3 Pathway.

BackgroundSeveral studies have indicated that the anoikis effector Bcl-2 inhibitor of transcription 1 (Bit1) can promote or inhibit tumor progression depending on the nature of the malignancy. However, its regulatory effects on gliomas are unknown.MethodsThis study aimed at assessing Bit1 expression in glioma tissues and cells, its subsequent effects on glioma cell apoptosis, proliferation, invasion, and migration, and the underlying molecular mechanisms.ResultsThe findings showed that lower Bit1 expressions in glioma tissues as well as a negative correlation between Bit1 expression and glioma grade. Additional findings also revealed that Bit1 silencing significantly inhibited anoikis and enhanced glioma cell proliferation, invasion, and migration. Further analysis showed that the decrease in Bit1 expressions led to malignancy proliferation and anoikis resistance through activation of the IL-6/STAT3 signaling pathway.ConclusionOur data suggested that Bit1 may play an anti-oncogenic role in glioma cells and that a decrease in its expressions might induce glioma cell proliferation, migration, and invasion through the IL-6/STAT3 signaling pathway.

A role of metallothionein-3 in radiation-induced autophagy in glioma cells

Although metallothionein-3 (MT3), a brain-enriched form of metallothioneins, has been linked to Alzheimer’s disease, little is known regarding the role of MT3 in glioma. As MT3 plays a role in autophagy in astrocytes, here, we investigated its role in irradiated glioma cells. Irradiation increased autophagy flux in GL261 glioma cells as evidenced by increased levels of LC3-II but decreased levels of p62 (SQSTM1). Indicating that autophagy plays a cytoprotective role in glioma cell survival following irradiation, measures inhibiting autophagy flux at various steps decreased their clonogenic survival of irradiated GL261 as well as SF295 and U251 glioma cells. Knockdown of MT3 with siRNA in irradiated glioma cells induced arrested autophagy, and decreased cell survival. At the same time, the accumulation of labile zinc in lysosomes was markedly attenuated by MT3 knockdown. Indicating that such zinc accumulation was important in autophagy flux, chelation of zinc with tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN), induced arrested autophagy in and reduced survival of GL261 cells following irradiation. Suggesting a possible mechanism for arrested autophagy, MT3 knockdown and zinc chelation were found to impair lysosomal acidification. Since autophagy flux plays a cytoprotective role in irradiated glioma cells, present results suggest that MT3 and zinc may be regarded as possible therapeutic targets to sensitize glioma cells to ionizing radiation therapy.

Long non-coding RNA SOX21-AS1 promotes cell proliferation and invasion through upregulating PAK7 expression by sponging miR-144-3p in glioma cells.

In this study, the function of long non-coding RNA SOX21 antisense RNA 1 (SOX21-AS1) in the progress of glioma was explored. RNA and protein levels were measured via quantitative reverse transcription-PCR (qRT-PCR) and western blot analysis. In addition, we examined cell proliferation, apoptosis, migration and invasion. The interaction between SOX21-AS1 (PAK7) and miR-144-3p was determined via RNA immunoprecipitation (RIP) assay and Luciferase reporter assay. SOX21-AS1 was upregulated in glioma tissues and cells. SOX21-AS1 knockdown was carried out in glioma cells (U251 and U87 cells). Moreover, in vitro, SOX21-AS1 knockdown repressed proliferation, migration, invasion and enhanced apoptosis in glioma cells. In vivo, SOX21-AS1 knockdown suppressed tumor growth in mice. In addition, SOX21-AS1 could sponge miR-144-3p, which was determined to bind to PAK7. miR-144-3p knockdown promoted proliferation, migration, invasion and inhibited cell apoptosis. Importantly, the effects of SOX21-AS1 knockdown-induced proliferation, migration, invasion, and apoptosis were alleviated in glioma cells co-transfected with SOX21-AS1 and miR-144-3p knockdown. Furthermore, miR-144-3p knockdown also attenuated Wnt/β-catenin pathway-associated protein levels induced by SOX21-AS1 knockdown. These results indicated that SOX21-AS1/miR-144-3p/PAK7 axis played an oncogenic role in glioma cells by regulating Wnt/β-catenin pathway, which suggests a rational therapeutic strategy for glioma.

MATN1-AS1 promotes glioma progression by functioning as ceRNA of miR-200b/c/429 to regulate CHD1 expression.

ObjectivesLong non‐coding RNA (lncRNA) MATN1‐AS1 is a newfound lncRNA that has been rarely explored in cancers. Herein, we would like to investigate its role in glioma.Materials and methodsqRT‐PCR was conducted to examine gene expression in glioma. Then, MTT assay, colony formation assay and flow cytometry analysis were applied to evaluate the function of MATN1‐AS1 on glioma cells. Western blot was performed to measure the protein levels of genes. Besides, the luciferase reporter assay, RNA pull‐down assay, RIP assay and Spearman's correlation analysis were also performed as needed.ResultsFirstly, a data from TCGA showed that MATN1‐AS1 might be largely implicated in glioma. Meanwhile, MATN1‐AS1 upregulation confirmed in glioma predicted poor clinical outcomes. Functionally, MATN1‐AS1 knockdown restrained cell proliferation but stimulated apoptosis in vitro and repressed tumour growth in vivo. Mechanistic investigations validated that MATN1‐AS1 functioned as a ceRNA for miR‐200b/c/429 to upregulate CHD1 which was also verified to exert a growth‐promoting role in glioma cells here. Importantly, both CHD1 overexpression and miR‐200b/c/429 inhibition could rescue the obstructive role of MATN1‐AS1 silence in glioma cells.ConclusionsMATN1‐AS1 promotes glioma progression through regulating miR‐200b/c/429‐CHD1 axis, suggesting MATN1‐AS1 as a probable target for glioma treatment.

The SIAH1-HIPK2-p53ser46 Damage Response Pathway is Involved in Temozolomide-Induced Glioblastoma Cell Death.

Patients suffering from glioblastoma have a dismal prognosis, indicating the need for new therapeutic targets. Here we provide evidence that the DNA damage kinase HIPK2 and its negative regulatory E3-ubiquitin ligase SIAH1 are critical factors controlling temozolomide-induced cell death. We show that HIPK2 downregulation (HIPK2kd) significantly reduces the level of apoptosis. This was not the case in glioblastoma cells expressing the repair protein MGMT, suggesting that the primary DNA lesion responsible for triggering HIPK2-mediated apoptosis is O6 -methylguanine. Upon temozolomide treatment, p53 becomes phosphorylated whereby HIPK2kd had impact exclusively on ser46, but not ser15. Searching for the transcriptional target of p-p53ser46, we identified the death receptor FAS (CD95, APO-1) being involved. Thus, the expression of FAS was attenuated following HIPK2kd, supporting the conclusion that HIPK2 regulates temozolomide-induced apoptosis via p-p53ser46-driven FAS expression. This was substantiated in chromatin-immunoprecipitation experiments, in which p-p53ser46 binding to the Fas promotor was regulated by HIPK2. Other pro-apoptotic proteins such as PUMA, NOXA, BAX, and PTEN were not affected in HIPK2kd, and also double-strand breaks following temozolomide remained unaffected. We further show that downregulation of the HIPK2 inactivator SIAH1 significantly ameliorates temozolomide-induced apoptosis, suggesting that the ATM/ATR target SIAH1 together with HIPK2 plays a proapoptotic role in glioma cells exhibiting p53wt status. A database analysis revealed that SIAH1, but not SIAH2, is significantly overexpressed in glioblastomas. IMPLICATIONS: The identification of a novel apoptotic pathway triggered by the temozolomide-induced DNA damage O6 -methylguanine supports the role of p53 in the decision between survival and death and suggests SIAH1 and HIPK2 as new therapeutic targets.

FXR1 promotes the malignant biological behavior of glioma cells via stabilizing MIR17HG

BackgroundAccumulating evidence has highlighted the potential role of RNA binding proteins (RBPs) in the biological behaviors of glioblastoma cells. Herein, the expression and function of RNA binding proteins FXR1 were investigated in human glioma cells.MethodsQuantitative real-time PCR were conducted to evaluate the expression of MIR17HG and miR-346, miRNA-425-5p in glioma tissues and cells. Western blot were used to explore the expression of FXR1, TAL1 and DEC1 in glioma tissues and cells. Stable knockdown of FXR1 and MIR17HG in glioma cells were established to explore the function of FXR1, MIR17HG in glioma cells. Further, RIP and RNA pull-down assays were used to investigate the correlation between FXR1 and MIR17HG. Cell Counting Kit-8, transwell assays, and flow cytometry were used to investigate the function of FXR1 and MIR17HG in malignant biological behaviors of glioma cells. ChIP assays were employed to ascertain the correlations between TAL1 and MIR17HG.ResultsFXR1and MIR17HG were upregulated in glioma tissues and cell lines. Downregulation of FXR1 or MIR17HG resulted in inhibition of glioma cells progression. We also found that FXR1 regulates the biological behavior of glioma cells via stabilizing MIR17HG. In addition, downregulated MIR17HG increased miR-346/miR-425-5p expression and MIR17HG acted as ceRNA to sponge miR-346/miR-425-5p. TAL1 was a direct target of miR-346/miR-425-5p, and played oncogenic role in glioma cells. More importantly, TAL1 activated MIR17HG promoter and upregulated its expression, forming a feedback loop. Remarkably, FXR1 knockdown combined with inhibition of MIR17HG resulted in the smallest tumor volumes and the longest survivals of nude mice in vivo.ConclusionsFXR1/MIR17HG/miR-346(miR-425-5p)/TAL1/DEC1 axis plays a novel role in regulating the malignant behavior of glioma cells, which may be a new potential therapeutic strategy for glioma therapy.

Mechanosensitivity of the BK Channels in Human Glioblastoma Cells: Kinetics and Dynamical Complexity

BK channels are potassium selective and exhibit large single-channel conductance. They play an important physiological role in glioma cells: they are involved in cell growth and extensive migrating behavior. Due to the fact that these processes are accompanied by changes in membrane stress, here, we examine mechanosensitive properties of BK channels from human glioblastoma cells (gBK channels). Experiments were performed by the use of patch-clamp method on excised patches under membrane suction (0–40 mmHg) at membrane hyper- and depolarization. We have also checked whether channel’s activity is affected by possible changes of membrane morphology after a series of long impulses of suction. Unconventionally, we also analyzed internal structure of the experimental signal to make inferences about conformational dynamics of the channel in stressed membranes. We examined the fractal long-range memory effect (by R/S Hurst analysis), the rate of changes in information by sample entropy, or correlation dimension, and characterize its complexity over a range of scales by the use of Multiscale Entropy method. The obtained results indicate that gBK channels are mechanosensitive at membrane depolarization and hyperpolarization. Prolonged suction of membrane also influences open–closed fluctuations—it decreases channel’s activity at membrane hyperpolarization and, in contrary, increases channel’s activity at high voltages. Both membrane strain and its “fatigue” reduce dynamical complexity of channel gating, which suggest decrease in the number of available open conformations of channel protein in stressed membranes.

ARTICLE WITHDRAWN] Knockdown of Long Noncoding RNA LINC00152 Suppresses Cellular Proliferation and Invasion in Glioma Cells by Regulating miR-4775.

Long noncoding RNAs (lncRNAs) play an important role in various biological properties of glioma cells. Herein we aimed to elucidate the function and the possible molecular mechanisms of long intergenic noncoding RNA 152 (LINC00152) in glioma cells. Relative expressions of LINC00152, miR-4775, and CDK6 in U-118 MG cells were regulated by transfections. Thereafter, cell viability, migration, invasion, and apoptosis were analyzed by CCK-8, Transwell, and flow cytometry assays. Dual-Luciferase Reporter Assay was conducted to validate the target genes of LINC00152 and miR-4775. Expression of components of the signal pathways were detected by Western blot. The results showed that LINC00152 knockdown significantly suppressed cell viability, migration, and invasion and induced apoptosis in vitro. Additionally, LINC00152 functioned as a molecular sponge for miR-4775, and inhibition of miR-4775 reversed the tumor-suppressive effects of LINC00152 knockdown on glioma cells. Furthermore, CDK6 was confirmed to be a target of miR-4775, and overexpression of CDK6 reduced apoptosis and abolished the inhibitory effects of miR-4775 overexpression on cell viability, migration, and invasion. Overexpression of CDK6 activated the PI3K/AKT/MAPK and Notch signal pathways. Overall, these findings demonstrate that LINC00152 plays an oncogenic role in glioma cells by regulation of miR-4775, which may therefore be a potential therapeutic target for glioma.

Prucalopride inhibits the glioma cells proliferation and induces autophagy via AKT-mTOR pathway

BackgroundsGlioma is the most fatal primary brain glioma in central nervous system mainly attributed to its high invasion. Prucalopride, a Serotonin-4 (5-HT4) receptor agonist, has been reported to regulate neurodevelopment. This study aimed to investigate the influence of the Prucalopride on glioma cells and unveil underlying mechanism.MethodsIn this study, glioma cells proliferation was evaluated by Cell counting kit-8 (CCK8). Wound healing and transwell assay were used to test cellular migration and invasion. Flow cytometry was utilized to determine cellular apoptosis rate. Apoptosis related markers, autophagy markers, and protein kinase B (AKT)-mammalian target of rapamycin (mTOR) pathway key molecules were detected using western blot assay.ResultsAs a result, the proliferation, migration and invasiveness of glioma cells were impaired by Prucalopride treatment, the apoptosis rate of glioma cells was enhanced by Prucalopride stimulation, accompanied by the increased pro-apoptosis proteins Bax and Cleaved caspase-3 and decreased anti-apoptosis protein Bcl-2. Prucalopride significantly promoted autophagy by increased expression level of Beclin 1 and LC3-II, while decreased expression level of p62. Prucalopride administration resulted in obvious inhibitions of key molecules of AKT-mTOR pathway, including phosphorylated- (p-) AKT, p-mTOR and phosphorylated-ribosomal p70S6 kinase (p-P70S6K).ConclusionsTaking together, these results indicate that Prucalopride may be likely to play an anti-tumor role in glioma cells, which suggests potential implications for glioma promising therapy alternation in the further clinics.

Forkhead Box A2 (FOXA2) Inhibits Invasion and Tumorigenesis in Glioma Cells.

The forkhead box A2 (FOXA2) is the key transcriptional factor that plays an important role in tumorigenesis. However, until now the expression pattern and role of FOXA2 in glioma have yet to be elucidated. Therefore, the aim of this study was to evaluate the expression of FOXA2 in glioma and investigate its role in glioma cells. Our data showed that FOXA2 was significantly downregulated in human glioma cell lines. Forced expression of FOXA2 suppressed the ability of glioma cells to proliferate, migrate, and invade and influenced the expression level of EMT-associated proteins. In addition, forced expression of FOXA2 attenuated tumor growth of glioma in a nude mouse xenograft model. Mechanistically, we disclosed that forced expression of FOXA2 greatly downregulated the expression of β-catenin, cyclin D1, and c-Myc in glioma cells. Taken together, these results show that FOXA2 may play an important role in proliferation, invasion, and tumorigenesis in glioma cells. Thus, FOXA2 may be a potential therapeutic target for the treatment of glioma.