Relationship Between Pericarotid Fat Density and Pathology-Based Carotid Plaque Risk Characteristics.

OBJECTIVES: In this study, the authors aimed to evaluate the relationship between pericarotid fat density (PFD) and pathologic carotid plaque risk characteristics. METHODS: The authors retrospectively evaluated 58 patients (mean age: 66.66 ± 7.26 y, 44 males) who were subjected to both carotid endarterectomy and carotid artery computed tomography angiography (CTA) at the authors' institution. The computed tomography values of the adipose tissue around the most severe stenosis carotid artery were measured, and the removed plaques were sent to the Department of Pathology for American Heart Association (AHA) classification. The Wilcoxon signed-rank test was used to detect the difference in PFD values between the operative and nonoperative sides. According to carotid plaque risk characteristics, the associations between PFD and 4 different risk characteristic subgroups were analyzed. The Student t test and χ2 test were used to compare differences between different risk subgroups. Receiver operating characteristic curve analysis was used to evaluate the predictive efficacy of PFD for carotid plaque risk characteristics. RESULTS: The operative side had higher mean Hounsfield units (HU) values compared with the nonoperative side (P < 0.001). The AHA VI and the intraplaque hemorrhage (IPH) subgroups had higher mean HU values compared with the non-AHA VI and the non-IPH subgroups (P < 0.05). Male patients presented with IPH more than female patients (P = 0.047). The results of receiver operating characteristic curve analysis showed that the mean HU value (operative side; area under the curve: 0.729, Sensitivity (SE): 59.26%, Specificity (SP): 80.65%, P = 0.003) had a certain predictive value for diagnosing high-risk VI plaques. Pericarotid fat density ≥ -68.167 HU is expected to serve as a potential cutoff value to identify AHA VI and non-AHA VI subgroups. CONCLUSION: PFD was significantly associated with vulnerable plaques, high-risk AHA VI plaques, and IPH, which could be an indirect clinical marker for vulnerable plaques.

MiR-137 targets and regulates E2F7 to suppress progression of glioma cells.

INTRODUCTION: The paper aimed to explore the mechanism of miR-137 in modulating glioma. MATERIAL AND METHODS: qRT-PCR detected miR-137 and E2F7 mRNA expression in cells. The protein expression of E2F7 was measured using Western blot assay. Cell proliferation, scratch healing, transwell and programmed cell death assays were conducted to examine the influences of the genes on the biological function of glioma cells. The dual-luciferase assay verified the interaction between miR-137 and E2F7. RESULTS: MiR-137 was lowly expressed in glioma cells, and E2F7 was highly expressed. MiR-137 suppressed progression and promoted programmed cell death of glioma cells. MiR-137 could target and negatively regulate E2F7 expression to further accelerate programmed cell death of glioma cells. CONCLUSIONS: It was found that miR-137 could target E2F7 to restrain cell progression and accelerate programmed cell death of glioma cells, which is helpful to search for new molecular therapeutic targets for glioma.

MicroRNA-433-3p enhances chemosensitivity of glioma to cisplatin by downregulating NR5A2.

OBJECTIVE: We attempted to investigate influence of microRNA-433-3p on malignant progression of glioma and identify its molecular mechanism, thus laying groundwork for glioma management. METHODS: Expression data along with clinical data of glioma were accessed from the TCGA database for differential and survival analyses to look for the target differentially expressed genes. Quantitative reverse transcriptase PCR (qRT-PCR) and western blot were utilized to assess NR5A2 mRNA and protein expression in different glioma cell lines, respectively. MTT, Transwell assay, and flow cytometry were carried out to assay the impact of NR5A2 on behaviors of glioma cells in vitro. Bioinformatics analysis was used to identify the upstream microRNA of NR5A2 in glioma, while dual-luciferase and western blot assays were used to detect binding of microRNA and NR5A2. Chemosensitivity of glioma cells was evaluated by cisplatin cytotoxicity test. RESULTS: NR5A2 was upregulated in both glioma tissues and cell lines. Dual-luciferase assay result showed binding site of microRNA-433-3p on NR5A2 mRNA 3'UTR, and microRNA-433-3p reduced NR5A2 expression. Cell assays revealed that silencing NR5A2 could hamper proliferation, invasion, and migration and enhance chemosensitivity to cisplatin while promoting glioma cell apoptosis and blocking glioma cells in G0/G1 phase. Rescue experiments also indicated that microRNA-433-3p suppressed glioma malignant progression via inhibiting NR5A2. CONCLUSION: MicroRNA-433-3p which is significantly poorly expressed in glioma targets NR5A2 to suppress glioma malignant progression and enhance chemosensitivity to cisplatin.

miR-218-5p inhibits the malignant progression of glioma via targeting TCF12.

Several studies have shown the ability of transcription factor 12 (TCF12) to promote tumor malignant progression, but its function in glioma cells has not been fully elucidated. In this study, we analyzed the data from TCGA by bioinformatics and found that in glioma tissue, TCF12 was conspicuously highly expressed while miR-218-5p was significantly low-expressed. The downregulation of miR-218-5p was correlated with adverse prognosis in patients with glioma. miR-218-5p was found to be negatively associated with TCF12 by Pearson correlation analysis, and dual luciferase assay was employed to verify that miR-218-5p and TCF12 had a targeting relationship. qRT-PCR and Western blot assays were used to verify that the expression of TCF12 was regulated by its upstream regulator miR-218-5p. Moreover, cell experiments validated that overexpressed TCF12 could promote the proliferation, migration, and invasion of glioma cells and inhibit their apoptosis, whereas overexpressing miR-218-5p at the same time could reverse this phenomenon. Our study demonstrates the regulatory mechanism of the miR-218-5p/TCF12 axis in gliomas, which lays a foundation for searching for new therapeutic approaches for glioma.

MiR-137 inhibits the proliferation, invasion and migration of glioma via targeting to regulate EZH2.

BACKGROUND: Gliomas are common malignant tumors in the nervous system, known for poor prognosis and low survival rate. OBJECTIVE: This study aims to explore functions of miR-137 in glioma progression and identify messenger RNAs (mRNA) regulated by miR-137, which provides new ideas for further exploration of glioma therapeutic targets. METHODS: Gene expression data were downloaded from the Cancer Genome Atlas database, and abnormally expressed miRNAs and mRNAs in glioma were analyzed. The expression of genes in 20 pairs of clinical tissue samples and glioma cell lines were detected through qRT-PCR, and the expression of proteins was detected through Western blot. Changes in cell proliferative level after transfection were detected via CCK8 assay, and changes in cell migratory and invasive abilities were detected by Transwell assay. Besides, dual-luciferase reporter assay was employed to testify binding relationship between two genes. RESULTS: Our study found that miR-137 was significantly and lowly expressed in glioma tissue and cell lines, and the prognoses of glioma patients with highly expressed miR-137 were more optimistic. Overexpressed miR-137 could remarkably inhibit proliferative, invasive and migratory abilities of glioma cells U87, while transfection of miR-137 inhibitor presented an opposite effect. Additionally, EZH2 was a direct target of miR-137 and overexpressed EZH2 effectively reversed the effect of miR-137 on glioma proliferation and migration. CONCLUSIONS: Our study found that miR-137 could suppress the proliferation, invasion and migration of glioma cells through regulating the expression of EZH2. So far, we have found a novel regulatory pair that influences glioma progression, providing a basis for further development of new therapeutic strategies.

MiR-433-3p restrains the proliferation, migration and invasion of glioma cells via targeting SMC4.

OBJECTIVE: Glioma is a common primary malignant brain tumor characterized by high mortality and poor prognosis. The purpose of this study is to explore the molecular mechanism underlying glioma, aiming to provide a new target for the treatment of glioma to improve the prognosis of patients. METHODS: The differentially expressed genes and regulatory axis affecting the prognosis of glioma were identified with bioinformatics analysis, and the expression of miR-433-3p and SMC4 mRNA was detected with qRT-PCR. The expression of SMC4 and epithelial-mesenchymal transition (EMT)-associated proteins were detected with western blot. The targeting relationship between miR-433-3p and SMC4 was verified with dual-luciferase reporter gene assay. The proliferative ability of glioma cells was detected with CCK-8 assay, while the migration and invasion of glioma cells were detected with Transwell assay. RESULTS: We found that the expression of SMC4 was significantly up-regulated in glioma, showing that SMC4 was an unfavorable factor for prognosis and could promote the progression of cancer cells. Its upstream regulator miR-433-3p was significantly down-regulated in glioma, which inhibited the development of cancer cells. Moreover, miR-433-3p could target to inhibit the expression of SMC4. Rescue assay showed that miR-433-3p could affect the development of glioma by regulating the expression of SMC4. CONCLUSION: Our data demonstrate for the first time that SMC4 is a direct target of miR-433-3p, and elucidate the molecular mechanism by which miR-433-3p inhibits the malignant progression of glioma by targeting and down-regulating the expression of SMC4.

HAS2-AS1 Acts as a Molecular Sponge for miR-137 and Promotes the Invasion and Migration of Glioma Cells by Targeting EZH2.

This study aims to explore the molecular mechanism by which HAS2-AS1 acts as a ceRNA to promote the invasion and migration of glioma cells, which will provide a novel potential target for the targeted therapy of glioma. Gene expression profiles and corresponding clinical data were accessed from the TCGA_LGG and TCGA_GBM databases and then differential analysis was conducted using the "edgeR" package. miRDB, miRTarBase and TargetScan databases were employed to predict target genes and sequentially a ceRNA network was constructed. Quantitative real-time PCR was performed to detect gene expression in glioma cells. Transwell assay was operated to assess cell migratory and invasive abilities. Western blot was conducted to evaluate the protein expression. Dual-luciferase reporter assay and RNA immunoprecipitation experiment were performed to validate the targeting relationship between genes. HAS2-AS1 was markedly upregulated in glioma, and the overall survival time of patients with high HAS2-AS1 expression was significantly shorter than that of patients with low one. Silencing HAS2-AS1 inhibited the migration and invasion of glioma cells, while overexpressing HAS2-AS1 produced opposite results. miR-137 was validated as a direct target of and negatively regulated by HAS2-AS1. Further exploration of the downstream target gene indicated that EZH2 competed with HAS2-AS1 to interact with miR-137. Suppressing miR-137 or up-regulating EZH2 reversed the impact of HAS2-AS1 knockdown on glioma cell invasion and migration. HAS2-AS1 regulates EZH2 by sponging miR-137 for the migratory and invasive abilities of glioma cells, which provides a new idea for exploring metastasis mechanism of glioma.

The transcription factor USF1 promotes glioma cell invasion and migration by activating lncRNA HAS2-AS1.

OBJECTIVE: The role of lncRNAs in tumor has been widely concerned. The present study took HAS2-AS1 (the antisense RNA 1 of HAS2) as a starting point to explore its expression in glioma and its role in the process of migration and invasion, providing a strong theoretical basis for mining potential therapeutic targets of glioma. METHODS: Clinical data of glioma were obtained from The Cancer Genome Atlas (TCGA) database and differentially expressed lncRNAs were analyzed by edgeR. The hTFtarget database was used to predict the upstream transcription factors of HAS2-AS1 and the JASPAR website was used to predict the binding sites of human upstream transcription factor 1 (USF1) and HAS2-AS1. qRT-PCR was used to detect the expressions of HAS2-AS1 and USF1 in glioma tissues and cell lines. The effects of silencing HAS2-AS1 on the migration and invasion of cancer cells were verified by wound healing and Transwell invasion assays. The chromatin immunoprecipitation (ChIP) and dual luciferase reporter assays were applied to demonstrate the binding of USF1 and HAS2-AS1 promoter region. Western blot was used to detect the expressions of epithelial-mesenchymal transition (EMT)-related proteins. RESULTS: HAS2-AS1 was highly expressed in glioma tissues and cells, and was significantly associated with poor prognosis. Silencing HAS2-AS1 expression inhibited glioma cell migration, invasion and EMT. USF1 was highly expressed in glioma and positively correlated with HAS2-AS1. The transcription of HAS2-AS1 was activated by USF1 via binding to HAS2-AS1 promoter region, consequently potentiating the invasion and migration abilities of glioma cells. CONCLUSION: These results suggested that the transcription factor USF1 induced up-regulation of lncRNA HAS2-AS1 and promoted glioma cell invasion and migration.