Levels In Glioma Tissues Research Articles

HMGB1 contributes to SASH1 methylation to attenuate astrocyte adhesion

SAM and SH3 domain-containing 1 (SASH1), a scaffold protein, is regarded as a tumor suppressor. Recent studies have verified the decreased expression of SASH1 in many tumors. Our previous clinical investigation found that SASH1 was widely expressed in normal brain tissues but reduced or absent in glioma tissues. However, the functions of SASH1 in normal astrocytes and the reasons for the reductions in SASH1 levels in glioma tissues are unclear. In this study, we found that in astrocytes, SASH1 functions in cell adhesion. We observed that knockdown of SASH1 expression in cultured astrocytes significantly decreased cell adhesion and increased invasion. Conversely, overexpression of SASH1 in C6 cells markedly promoted cell adhesion and decreased cell invasion. In addition, we found that the expression level of one member of the integrin family, integrin β8, was significantly reduced in SASH1-downregulated astrocytes and elevated in SASH1-upregulated C6 cells. Furthermore, the results of methylation and ChIP assays showed that the methylation level of the SASH1 gene was markedly higher in C6 cells than in astrocytes and that HMGB1 could bind to the CpG islands of the SASH1 gene. HMGB1 overexpression in astrocytes significantly increased the methylation level of the SASH1 gene. This study reveals, for the first time, that HMGB1 contributes to the methylation of the SASH1 gene, and our findings suggest that methylation downregulates the expression of the SASH1 gene and later reduces integrin β8 expression, thereby reducing cell adhesion and promoting cell migration.

TRIM28 is overexpressed in glioma and associated with tumor progression.

BackgroundTripartite motif containing 28 (TRIM28) is a transcriptional co-factor targeting many genes with pleiotropic biological activities, but the study on the role of TRIM28 in glioma is rare.MethodsTo explore the function of TRIM28 in glioma, we first detected the expression levels of TRIM28 in glioma tissues and analyzed the correlations of TRIM28 expression with clinicopathological variables of patients in 85 cases of glioma. Meanwhile, we used shRNA to knockdown TRIM28 in glioma cell lines to detect the biological functions of TRIM28 in cell and animal experiments.ResultsWe found that TRIM28 was expressed at significantly higher level in glioma tissues than in non-tumor brain, and TRIM28 expression correlated significantly with tumor malignancy. Furthermore, TRIM28 higher expression was also correlated with poor survival of glioma patients (P<0.01). Functionally, knockdown of TRIM28 could significantly inhibit cell proliferation and migration in glioma cells. Additionally, we found that TRIM28 could inhibit the expression of E-cadherin significantly by reducing its mRNA stability at the post-transcriptional level.ConclusionOur results suggest that TRIM28 overexpression is correlated with glioma malignant progression and patients’ poor survival, so targeting TRIM28 could be an efficacious strategy in glioma.

Ketohexokinase is involved in fructose utilization and promotes tumor progression in glioma

Many studies have pinpointed that fructose could be utilized as a carbon source in some cancers, and we also defined that glioma cells could utilize fructose to maintain themselves survival and proliferation depending on GLUT5 expression recently. However, ketohexokinase (KHK), as a key enzyme involved in fructose catabolism, drew less attention in cancers, especially in glioma. In the present study, we first analyzed the expression levels of KHK in glioma tissues and the correlations of KHK expression with clinicopathological variables of patients with glioma. Meanwhile, we detected the effect of silencing KHK on the biological functions of glioma cells in fructose medium. From the results, we found that KHK was expressed at significantly higher level in glioma tissues than in non-tumor brain, and KHK expression was significantly correlated with tumor malignancy and poor survival of glioma patients (p < 0.01). Functionally, knockdown of KHK could significantly inhibit cell proliferation and migration of glioma cells in fructose medium. Furthermore, we investigated the KHK expression level after long-time treatment with fructose, and detected the change of cell biological behaviour, then we found that the expression level of KHK was significantly increased and these cells showed more malignant properties. Taken together, our results suggest that high fructose diet and KHK overexpression are correlated with glioma malignant progression and patients' poor survival, and we believe this hypothesis would open the door for novel therapeutic agents and mentalities for glioma.

Ten-eleven translocation 1 regulates methylation of autophagy-related genes in human glioma.

Ten-eleven translocation 1 catalyzes the conversion of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), which plays an important role in epigenetics and is related to the malignant biological behavior of tumors. However, its regulatory role in glioma remains unclear. In this study, the levels of 5mC and 5hmC were detected using immunohistochemistry, dot-blot, hMeDIP-chip, and western blot in glioma tissues and normal brain tissues, whereas 5hmC differentially enriched genes were determined and further validated. The level of 5hmC in gliomas was decreased, whereas 5mC was increased. 5hmC highly enriched 10 functional protein-coding genes and 10 signaling pathways were identified using hMeGIP-chip in glioma tissues. Two autophagy-related genes, ATG13 and DNA damage-regulated autophagy modulator protein 1, with low enrichment of 5hmC in glioma tissues were verified in the promoter region, and hMeGIP-PCR further confirmed this result in U251 cells. Immunohistochemistry further confirmed that autophagy level in glioma tissues was lower than that of normal controls, and negatively correlated with WHO grade. This study indicates that ten-eleven translocation 1 may be involved in the development and progression of glioma through demethylation regulating a variety of cellular functions and signaling pathways, and autophagy is one of the regulatory mechanisms.

MicroRNA-599 suppresses glioma progression by targeting RAB27B.

MicroRNAs (miRNAs/miRs) serve tumor promoting or suppressive roles in different human cancer types, including glioma; however, the regulatory underlying mechanism by which miR-599 affects glioma progression remains largely unknown. The aim of the present study was to investigate the expression of miR-599 in glioma, as well as the underlying regulatory mechanism. Reverse transcription-quantitative polymerase chain reaction and western blot analysis were used to examine mRNA and protein expression, respectively. MTT, wound healing and transwell assays were conducted to study cell proliferation, migration and invasion, respectively. A dual-luciferase reporter gene assay was used to confirm the targeting association between miR-599 and Ras-related protein Rab-27B (hereafter RAB27B). In the present study, miR-599 expression was observed to be significantly downregulated in human glioma tissues and cell lines, when compared with normal brain tissues and normal human astrocyte cells, respectively. Low miR-599 expression was significantly associated with glioma progression. Ectopic expression of miR-599 caused a significant reduction in the proliferation, migration and invasion of U-87MG Uppsala and U251 cells. Bioinformatics analysis and dual-luciferase reporter gene assay data identified that RAB27B was a direct target gene of miR-599. The mRNA and protein expression of RAB27B was significantly downregulated following miR-599 overexpression in U-87MG Uppsala and U251 cells. Rescue experiments demonstrated that RAB27B rescued the miR-599-induced inhibition of glioma cell growth, migration and invasion. In addition, RAB27B expression was significantly upregulated in glioma tissues and cell lines, and the expression levels of RAB27B were inversely correlated with miR-599 levels in glioma tissues; therefore, the present study demonstrated that miR-599 exerts a tumor-suppressive role in glioma progression by targeting RAB27B, indicating that miR-599 may be used as a potential candidate for glioma treatment.

MicroRNA-423-3p promotes glioma growth by targeting PANX2.

Previously, a number of microRNAs (miRs) have been identified to participate in the development and progression of glioma via the regulation of their target genes. However, the molecular mechanisms underlying the effect of miR-423-3p in glioma growth remain unclear. In the present study, the reverse transcription-quantitative polymerase chain reaction and western blotting were used to assess the mRNA and protein expression levels of miR-423-3p, respectively. An MTT assay and flow cytometry were performed to determine cell proliferation and apoptosis, respectively. A luciferase reporter gene assay was performed to determine the target association between pannexin 2 (PANX2) and miR-423-3p. It was revealed that miR-423-3p was significantly upregulated in glioma tissues compared with normal brain tissues, and the increased expression of miR-423-3p was significantly associated with an advanced grade as well as a poorer prognosis of patients with glioma. Inhibition of miR-423-3p using an miR-423-3p inhibitor resulted in the decreased proliferation of glioma U251 and U87MG Uppsala cells, and the induction of apoptosis. PANX2 was identified as a novel target gene of miR-423-3p, and the expression of PANX2 was revealed to be increased in U251 and U87MG Uppsala cells when miR-423-3p was inhibited. Knockdown of PANX2 attenuated the effects of miR-423-3p inhibition on glioma cell proliferation and apoptosis. Furthermore, PANX2 was significantly downregulated in glioma tissues compared with normal brain tissues, and its levels were markedly lower in World Health Organization (WHO) stage III–IV gliomas compared with WHO stage I–II gliomas. Additionally, the expression levels of PANX2 were identified to be inversely correlated with miR-423-3p expression levels in glioma tissues. Consequently, targeting miR-423-3p may inhibit glioma growth via the upregulation of PANX2.

MiR-129-5p inhibits glioma cell progression invitro and invivo by targeting TGIF2.

This study purposed to explore the correlation between miR‐129‐5p and TGIF2 and their impacts on glioma cell progression. Differentially expressed miRNA was screened through microarray analysis. MiR‐129‐5p expression levels in glioma tissues and cells were measured by qRT‐PCR. CCK‐8 assay, flow cytometer, transwell assay and wound‐healing assay were employed to detect cell proliferation, apoptosis and cycle, invasiveness and migration, respectively. Dual‐luciferase reporting assay was performed to confirm the targeted relationship between miR‐129‐5p and TGIF2. The effects of TGIF2 expression on cell biological functions were also investigated using the indicated methods. Tumour xenograft was applied to explore the impact of miR‐129‐5p on tumorigenesis in vivo. MiR‐129‐5p expression was down‐regulated in both glioma tissues and glioma cells, while TGIF2 expression was aberrantly higher than normal level. Dual‐luciferase reporter assay validated the targeting relation between miR‐129‐5p and TGIF2. Overexpression of miR‐129‐5p or down‐regulation of TGIF2 inhibited the proliferation, invasion and migration capacity of glioma cells U87 and U251, and meanwhile blocked the cell cycle as well as induced cell apoptosis. MiR‐129‐5p overexpression repressed the tumour development in vivo. MiR‐129‐5p and TGIF2 had opposite biological functions in glioma cells. MiR‐129‐5p could inhibit glioma cell progression by targeting TGIF2, shining light for the development of target treatment for glioma.

MicroRNA-103 suppresses glioma cell proliferation and invasion by targeting the brain-derived neurotrophic factor.

Glioma is the most common and aggressive of malignant brain tumours. MicroRNAs (miRNAs/miRs) are involved in tumour development of various human cancers, including glioma. Therefore, miRNAs may have potential tumour diagnostic, prognostic and therapeutic values in human glioma. miR‑103 is abnormally expressed in various human cancer types. However, the detailed expression pattern, biological functions and underlying molecular mechanism of miR‑103 in glioma remain unclear. Therefore, the present study aimed to investigate the expression, biological roles and underlying mechanisms of miR‑103 in glioma. Results of the present study demonstrated that miR‑103 was significantly down‑regulated in glioma tissues and cell lines. Functional experiments demonstrated that miR‑103 overexpression inhibited the proliferation and invasion of glioma cells invitro. Additionally, brain‑derived neurotrophic factor (BDNF) was identified as a direct functional target of miR‑103 in glioma. Furthermore, mRNA and protein expression levels of BDNF were highly upregulated in glioma tissues compared with normal brain tissues. Spearman's correlation analysis indicated a negative association between miR‑103 and BDNF mRNA expression levels in glioma tissues. Furthermore, rescue experiments demonstrated that BDNF up‑regulation reversed the suppressive effects of miR‑103 on glioma cell proliferation and invasion. Therefore, the authors of the present study hypothesized that the interaction between miR‑103 and BDNF serves a role in glioma progression and, in the future, may serve as a therapeutic target for glioma treatment.

Expression and clinical significance of cyclooxygenase 2 and survivin in human gliomas.

The present study aimed to determine cyclooxygenase 2 (COX-2) and survivin expression levels in glioma tissues, and to investigate their association with clinicopathological factors and patient survival. Immunohistochemistry was performed to evaluate COX-2 and survivin expression levels in paraffin-embedded surgically resected tissues from 70 patients with glioma and 7 individuals with normal brain tissues. The association between COX-2 and survivin expression levels and clinicopathological features was investigated using the χ2 test, and the survival time was analyzed using the Kaplan Meier method with log-rank test. COX-2 and survivin were overexpressed in glioma tissues, and higher expression levels were observed in glioma tissues of histological grades III–IV compared with those in grade I–II tumor tissues (P<0.05); however, the expression levels were not associated with gender, age, tumor size or location (P>0.05). There was a significant positive association between the expression levels of COX-2 and survivin in the glioma tissues. Additionally, COX-2 and survivin expression levels were significantly negatively correlated with the rate of survival. In conclusion, COX-2 and survivin expression is positively associated with the pathological grade of a glioma and may contribute to glioma tumorigenesis. Therefore, COX-2 and survivin may be sensitive predictors of a negative clinical prognosis for patients with glioma.

Serum immunoreactivity of cancer/testis antigen OY-TES-1 and its tissues expression in glioma.

OY-TES-1 is a member of the cancer/testis antigen family that is expressed in healthy testis tissue and certain types of cancerous tissue. The present study aimed to analyze the expression pattern of OY-TES-1 and serum anti-OY-TES-1 antibody concentration in patients with glioma. OY-TES-1 mRNA was detected in 28/36 (78%) of glioma cases using conventional reverse transcription polymerase chain reaction (RT-PCR) analysis. RT-quantitative-PCR revealed that OY-TES-1 was expressed at a higher level in glioma tissues compared with normal adult tissues (with the exception of testis tissue). Anti-OY-TES-1 antibodies were present in the serum of 5/36 (14%) of patients with glioma, but absent in all the serum samples from 107 healthy donors. Immunohistochemical analysis demonstrated that OY-TES-1 protein was expressed in all glioma tissues from patients with anti-OY-TES-1 antibody seropositivity. These results suggest that OY-TES-1 is a novel candidate for glioma immunotherapy.

IQGAP1‑siRNA inhibits proliferation and metastasis of U251 and U373 glioma cell lines.

IQ motif containing GTPase activating protein 1 (IQGAP1) is a scaffold protein, which is aberrantly expressed in several tumor types and is closely associated with the development, metastasis and prognosis of cancer. Several studies have demonstrated that IQGAP1 has broad prospects in the basic and clinical research of tumors. The present study aimed to explore the effects of IQGAP1-small interfering (si) NA on the proliferation and metastasis of U251 and U373 glioma cell lines, which markedly expressed IQGAP1. The human glioma cell lines (U251 and U373) were transfected with siRNA and transfection efficacy was confirmed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. Cell proliferation was detected using the Cell Counting kit-8, and cell metastasis capabilities were detected using cell adhesion, migration and invasion assays. In addition, the expression levels of several tumor-associated genes were determined by RT-qPCR and western blotting. The results indicated that IQGAP1 was expressed at higher levels in glioma tissues compared with in normal brain tissues. IQGAP1-siRNA significantly inhibited cell proliferation, and cell adhesion, migration and invasion. Furthermore, the expression levels of matrix metalloproteinase (MMP)2, Snail, MMP9, fibronectin 1 and Twist were suppressed, and E-cadherin was upregulated in response to siRNA-IQGAP1. The present study identified the function of IQGAP1 in glioma cell biology, and indicated that it may be considered a novel target for glioma treatment.

MiRNA‑1271 inhibits cell proliferation in neuroglioma by targeting fibronectin 1.

miR-1271 is a multifunctional post-translational modulator, which is involved in several diseases. However, the association between microRNA (miR)-1271 and fibronectin 1 (FN1) remains to be fully elucidated in neuroglioma. In the present study, it was hypothesized that a post-translational mechanism of miR-1271 regulates the expression of FN1 in the progression of neuroglioma. The present study aimed to investigate the clinical significance and underlying molecular mechanisms of miRNA-1271 in the development of glioma. The miR-1271 levels in glioma tissues and cell lines were assessed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). miR-1271 mimics and inhibitors were transfected to gain or loss of miR-1271 function. Cell proliferation was analyzed by using an MTT assay. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. The mRNA and protein levels were assessed by RT-qPCR and western blotting. The results showed that miR-1271 was downregulated in glioma tumor tissues and cell lines. In addition, it was demonstrated that low levels of miR-1271 in patients with glioma were correlated with low survival rate. In vitro, the cell viability was significantly suppressed following transfection with miRNA-1271 mimics and increased following transfection with the miRNA-1271 inhibitor. The miRNA-1271 mimics induced cell apoptosis and the miRNA-1271 inhibitor suppressed cell apoptosis in H4 and U251 cell lines. Furthermore, the 3′-untranslated region of FN1 was bound by miR-1271. Therefore, it was concluded that miR-1271 inhibited glioma cell growth by targeting FN1, and a low level of miR-1271 in glioma tumor tissues was associated with lower survival rates in patients with glioma.

MicroRNA-16 inhibits the proliferation, migration and invasion of glioma cells by targeting Sal-like protein4.

MicroRNAs (miRNAs or miRs), a class of non-coding RNAs 18-25nucleotides in length, act as key regulators in the development and malignant progression of various human cancers by modulating the expression of their target genes. Recently, miR‑16 has been demonstrated to be play a role in glioma. However, the regulatory mechanisms of miR‑16 in glioma growth and metastasis remain largely unclear. In the present study, qRT-PCR revealed that miR‑16 was significantly downregulated in 23glioma tissue specimens compared to 7normal brain tissue specimens. Moreover, its levels were markedly lower in the glioma samples at stagesT2-T4 compared to those at stageT1. The overexpression of miR-16 significantly suppressed the proliferation, migration and invasion of U251 and U87glioma cells. Luciferase reporter assay identified Sal-like protein4(SALL4) as a target gene of miR‑16, and its protein levels were found to be decreased in miR‑16-overexpressing U251 and U87cells. Furthermore, the overexpression of SALL4 significantly reversed the suppressive effects of miR‑16 on the proliferation, migration and invasion of U251 and U87 cells, suggesting that miR‑16 playsa tumor suppressor role in glioma by inhibiting cell proliferation and invasion through the targeting of SALL4. Finally, we found that SALL4 was significantly upregulated in glioma tissues compared to normal brain tissues, and its levels were markedly higher in the glioma tissues at stagesT2-T4 compared to those at stageT1. In addition, the expression levels of SALL4 inversely correlated with the miR‑16 levels in glioma tissues, suggesting that the downregulation of miR‑16 contributes to the upregulation of SALL4 in glioma. On the whole, the findings of this study indicate a role for the miR‑16/SALL4 axis in glioma. Our data may also provide a potential therapeutic target for the treatment of glioma.

Overexpression of transient receptor potential mucolipin-2 ion channels in gliomas: role in tumor growth and progression.

The Transient Receptor Potential (TRP) superfamily consists of cation-selective and non-selective ion channels playing an important role both in sensory physiology and in physiopathology in several complex diseases including cancers. Among TRP family, the mucolipin (TRPML1, −2, and −3) channels represent a distinct subfamily of endosome/lysosome Ca2+ channel proteins. Loss-of-function mutations in human TRPML-1 gene cause a neurodegenerative disease, Mucolipidosis Type IV, whereas at present no pathology has been associated to human TRPML-2 channels.Herein we found that human TRPML-2 is expressed both in normal astrocytes and neural stem/progenitor cells. By quantitative RT-PCR, western blot, cytofluorimetric and immunohistochemistry analysis we also demonstrated that TRPML-2 mRNA and protein are expressed at different levels in glioma tissues and high-grade glioma cell lines of astrocytic origin. TRPML-2 mRNA and protein levels increased with the pathological grade, starting from pylocitic astrocytoma (grade I) to glioblastoma (grade IV). Moreover, by RNA interference, we demonstrated a role played by TRPML-2 in survival and proliferation of glioma cell lines. In fact, knock-down of TRPML-2 inhibited the viability, altered the cell cycle, reduced the proliferation and induced apoptotic cell death in glioma cell lines. The DNA damage and apoptosis induced by TRPML-2 loss increased Ser139 H2AX phosphorylation and induced caspase-3 activation; furthermore, knock-down of TRPML-2 in T98 and U251 glioma cell lines completely abrogated Akt and Erk1/2 phosphorylation, as compared to untreated cells.Overall, the high TRPML-2 expression in glioma cells resulted in increased survival and proliferation signaling, suggesting a pro-tumorigenic role played by TRPML-2 in glioma progression.

MiR-133b inhibits glioma cell proliferation and invasion by targeting Sirt1.

MicroRNAs (miRs) are a class of small non-coding RNAs that function as mediators of gene expression. Dysregulations of miRs have been implicated in the development and progression of glioma. In the present study, we investigated the role of miR-133b in mediating the proliferation and invasion of glioma cells, and the potential mechanism. Real-time RT-PCR results showed that miR-133b expression was significantly decreased in glioma tissues compared with normal brain tissues. Luciferase reporter assay further identified silent information regulator 1 (Sirt1) as a novel direct target of miR-133b in glioma U87 cells. Overexpression of miR-133b suppressed Sirt1 expression and reduced the proliferation and invasion of U87 cells, which could be partly rescued by forced expression of Sirt1. In addition, the Sirt1 mRNA level was significantly higher in glioma tissues than in normal brain tissues, and was inversely correlated with miR-133b level in glioma tissues. In summary, our study sheds light on the regulatory mechanism of miR-133b in glioma growth and metastasis via direct mediation of Sirt1 expression, and suggests that Sirt1 may serve as a potential therapeutic target for glioma.

The expression of miR-21 in brain glioma cells and its effect of PI3K/AKT signal pathway Running title: MiR-21 in glioma

Background:Brain glioma is a common primary intracranial tumor with unfavorable prognosis. MicroRNA-21 (miR-21) is a small RNA molecule involved in post-transcriptional modulation and is closely associated with cell growth and differentiation. Recent studies have shown that miR-21 expression affects tumor proliferation and growth via the PI3K/AKT signaling pathway. This study investigated the expression of miR-21 in glioma and its effects on the PI3K/AKT pathway and cell proliferation. Methods: A total of 35 brain glioma patients were recruited from March 2013 to November 2014 at our hospital. In situ hybridization (ISH) and RT-PCR were used to detect the expression of miR-21 in both tumors and adjacent brain tissues. Human brain U251 glioma cells were transfected with anti-miR-21 to suppress miR-21 expression, followed by immunofluorescent assay of PI3K and AKT expression profiles. Flow cytometry was employed to detect the change between different stages of the cell cycle. Results:ISH results showed significantly elevated miR-21 expression level in glioma tissues compared to normal brain tissues. RT-PCR analysis revealed a 3.5-fold increase in miR-21 in the anti-miR-21 transfected glioma cells compared to untransfected cells. After silencing miR-21 in U251 cells, immunofluorescent assay indicated that the expression of PI3K and AKT significantly decreased. Flow cytometry revealed an increased number of cells at G0/G1 phase in anti-miR-21 transfected U251 cells, suggesting suppressed cell division ability. Conclusion:MiR-21 expression was found to be increased in brain glioma tissues, accompanied with over-activation of PI3K/AKT signaling and proliferation of glioma cells

Overexpression of interleukin-33 is associated with poor prognosis of patients with glioma

Interleukin-33 (IL-33) has shown diagnostic and prognostic values in multiple human cancers. However, there is little knowledge on the role of IL-33 in human gliomas and its association with disease prognosis. This study aimed to evaluate the value of IL-33 in the prognosis of glioma patients. The expression of IL-33 was determined and compared in surgical specimens from 86 glioma patients and 16 normal brain tissues. The associations of IL-33 expression with the clinicopathological features and prognosis of glioma patients were assessed. qRT-PCR assay showed higher IL-33 mRNA expression in glioma tissues than in normal brain tissue ( p < 0.001), and significantly higher IL-33 mRNA expression was detected in both low- and high-grade glioma tissues relative to normal brain tissues ( p < 0.001). Western blotting revealed elevated IL-33 protein levels in glioma tissues compared to those in normal brain tissues, and immunohistochemical staining showed higher IL-33 protein expression in glioma tissues than in normal brain tissues. IL-33 expression correlated with the glioma grade ( p < 0.001) and Karnofsky performance status score ( p = 0.024), and the glioma patients with high IL-33 expression had a shorter progression-free survival ( p < 0.001) and overall survival ( p < 0.001) than those with low IL-33 expression. The univariate and multivariate analyses showed that IL-33 overexpression and the glioma grade were independent factors of a poor prognosis in glioma patients. Therefore, IL-33 may be a promising biomarker for the detection of gliomas, and IL-33 expression is useful for predicting the prognosis of the disease.

Low p21 level is necessary for the suppressive effects of micoRNA-31 on glioma cell migration and invasion.

MicroRNAs (miRNAs), a kind of endogenous non-coding RNAs, regulate gene expression through binding to the 3'-untranslational region (UTR) of target messenger RNAs (mRNAs) and act as endogenous agents of RNA interference, resulting in either mRNA degradation or translational repression. MiR-31 has been demonstrated to be associated with the development and progression of glioma. However, the underlying molecular mechanism remains largely unclear. In the present study, we demonstrated that miR-31 only inhibited the cell migration and invasion, as well as the expression of a known miR-31 target oncogene radixin, in U251 glioma cells that expressed low level of p21; however, miR-31 showed no above effects on glioma SHG44 cells that highly expressed p21. Moreover, upregulation of p21 in U251 cells reversed the suppressive effects of miR-31 on the cell migration and invasion, suggesting that low p21 level is necessary for the miR-31-mediated inhibitory effects on glioma. Furthermore, analysis for 35 glioma specimens showed that the expression of radixin was negatively correlated with the miR-31 level in glioma tissues with low p21 expression; however, no such correlation was found in glioma tissues with high p21 level, further supporting that the low p21 level is necessary for the suppressive effect of miR-31 on the expression of its target oncogenes. In summary, our study demonstrates that the suppressive effect of miR-31 on glioma cell migration and invasion is p21-dependent, and suggests that miR-31 may be used for the treatment of patients with p21-deficent glioma.

MiR-144-3p exerts anti-tumor effects in glioblastoma by targeting c-Met.

The study aimed to explore the specific function and mechanism of miR-144-3p in glioblastoma (GBM) cells with different phosphatase and tensin homolog (PTEN) phenotypes. We demonstrated that the miR-144-3p level was significantly down-regulated in glioma compared with the non-neoplastic brain tissues, and decreased with ascending grades. The loss of miR-144-3p effectively predicted the decreased overall survival in glioma patients. Interestingly, the expression of MET was up-regulated and inversely associated with miR-144-3p level in glioma tissues. Next, we certified that miR-144-3p specifically bound to MET 3'-untranslated region (3' UTR) and inhibited its expression. miR-144-3p potently repressed GBM cell proliferation and invasion via suppressing MET in vitro and in vivo. In addition, our results showed no difference in malignancy inhibition induced by miR-144-3p in GBM cells with different PTEN phenotypes. miR-144-3p inhibited several survival signaling pathways by targeting MET independent of PTEN status in GBM cells. Over-expression of miR-144-3p inhibited survival capability and increased apoptosis, resulting in enhancement of radiation and temozolomide sensitivity. Our data provide new insights into the potential application of miR-144-3p in GBM therapy by targeting MET and then inhibiting the downstream signaling.

Expression of CPEB4 in Human Glioma and Its Correlations With Prognosis.

CPEB4 plays an important role in cancer progression. However, the clinicopathological significance of CPEB4 expression to glioma and its expression levels in glioma tissues and cell lines are unknown. The present study investigated the potential prognostic value of CPEB4 for human glioma.Immunohistochemistry (IHC) was performed to examine the dynamics of CPEB4 expression in glioma and nonneoplastic brain tissues, and the expression of CPEB4 in cell lines and freshly prepared tissue samples was measured using Western blotting and real-time PCR.CPEB4 was highly expressed at the mRNA and protein levels in 4 glioma cell lines and in 4 freshly prepared glioma tissues. Immunohistochemical analysis demonstrated that CPEB4 expression in glioma tissue was higher than that in corresponding nonneoplastic brain tissue (P < 0.01). This high expression level was further increased in high-grade gliomas, and the CPEB4 expression level correlated with the WHO classification (r = 0.774, P < 0.01). Moreover, the overall survival of glioma patients displaying high CPEB4 protein expression (P < 0.01) was clearly lower than that of those displaying low CPEB4 expression, and the high CPEB4 expression indicated a poorer survival in high-grade glioma patients (P < 0.01).Our study suggests that CPEB4 is significantly expressed in human glioma and that the upregulation of CPEB4 protein is significantly associated with advanced WHO grade. CPEB4 may serve as a highly sensitive prognostic indicator for glioma patients.