sincalide and Brain-Neoplasms

The prognosis for the WHO grade 4 IDH-mutant astrocytoma is better than IDH-wildtype glioblastoma (GBM) patients. The purpose of this study is to explore the potential mechanism of how IDH1 mutation can increase the efficacy of radiotherapy and to establish a risk-score model to predict the efficacy of radiotherapy in WHO grade 4 gliomas. First, we conducted experimental study on the effect of IDH1

Topics: Brain Neoplasms; Calmodulin-Binding Proteins; Glioblastoma; Glioma; Humans; Isocitrate Dehydrogenase; Mutation; Prognosis; Sincalide; World Health Organization

Previous studies have suggested an important role for N6-methyladenosine (m6A) modification in the proliferation of glioma cells. N6, 2'-O-dimethyladenosine (m6Am) is another methylated form affecting the fate and function of most RNA. PCIF1 has recently been identified as the sole m6Am methyltransferase in mammalian mRNA. However, it remains unknown about the role of PCIF1 in the growth and survival of glioma cells.. We constructed glioma cell lines that stably downregulated/upregulated PCIF1, established intracranial xenograft models using these cell lines, and employed the following methods for investigations: CCK-8, EdU, colony formation, flow cytometry, qRT-PCR, Western blot, and immunohistochemistry.. Downregulating PCIF1 promoted glioma cell proliferation, while overexpressing PCIF1 showed the opposite effects. Overexpression of PCIF1 blocked cell cycle progression and induced apoptosis in glioma cells, which was further confirmed by alterations in the expression of cell checkpoint proteins and apoptotic markers. Interestingly, disruption of PCIF1 methyltransferase activity slightly reversed the effect of PCIF1 overexpression on cell proliferation, but had no significant reversal effects on cell cycle progression or apoptosis. Knockdown of PCIF1 promoted the growth of gliomas, while overexpressing PCIF1 inhibited tumor growth and prolonged the survival time of tumor-bearing mice. In addition, the mRNA and protein levels of PCIF1 were gradually decreased with the increase of WHO grade in glioma tissues, but there was no significant correlation with patient survival.. These results indicated that PCIF1 played a suppressing role in glioma growth and survival, which may not entirely depend on its methyltransferase activity.

Topics: Adaptor Proteins, Signal Transducing; Animals; Brain Neoplasms; Cell Proliferation; Glioma; Humans; Mammals; Methyltransferases; Mice; Nuclear Proteins; RNA, Messenger; Sincalide

Echinacoside (ECH) is a phenylethanoid extracted from the stems of Cistanches salsa, an herb used in Chinese medicine formulations, and is effective against glioblastoma multiforme (GBM). Epithelial-mesenchymal transition (EMT) is the cornerstone of tumorigenesis and metastasis, and increases the malignant behavior of GBM cells. The S phase kinase-related protein 2 (skp2), an oncoprotein associated with EMT, is highly expressed in GBM and significantly associated with drug resistance, tumor grade and dismal prognosis. The aim of this study was to explore the inhibitory effects of ECH against GBM development and skp2-induced EMT.. CCK-8, EdU incorporation, transwell, colony formation and sphere formation assays were used to determine the effects of ECH on GBM cell viability, proliferation, migration and invasion in vitro. The in vivo anti-glioma effects of ECH were examined using a U87 xenograft model. The expression levels of skp2 protein, EMT-associated markers (vimentin and snail) and stemness markers (Nestin and sox2) were analyzed by immunofluorescence staining and western blotting experiments.. ECH suppressed the proliferation, invasiveness and migration of GBM cells in vitro, as well as the growth of U87 xenograft in vivo. In addition, ECH downregulated the skp2 protein, EMT-related markers (vimentin and snail) and stemness markers (sox2 and Nestin). The inhibitory effects of ECH were augmented in the skp2-knockdown GBM cells, and reversed in cells with ectopic expression of skp2.. ECH inhibits glioma development by suppressing skp2-induced EMT of GBM cells.

Topics: Brain Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Glioblastoma; Glioma; Glycosides; Humans; Nestin; S-Phase Kinase-Associated Proteins; Sincalide; Vimentin

MicroRNAs (miRNAs) are a class of small noncoding RNAs that play important roles in tumor development and progression. miR-20a acts as an oncogene in many cancers; however, the underlying role of miR-20a in human glioma remains unknown.. Glioma tissue samples were obtained from 32 patients with primary glioma who had undergone surgery at the First Affiliated Hospital of Sun Yat-sen University (Guangzhou, China). Twenty-two normal brain tissue samples used as controls were obtained by internal decompression in patients who had undergone surgery for cerebral injury and cerebral hemorrhage at the same hospital.. Quantitative reverse transcription polymerase chain reaction showed upregulation of miR-20a in glioma tissues and cell lines compared with normal brain tissue and normal human astrocytes. Functional assays showed that miR-20a promotes proliferation and invasion and inhibits apoptosis in glioma cells. The bioinformatic analysis showed that CELF2 (CUGBP Elav-like family member 2) is a direct target gene of miR-20a, which was confirmed using a luciferase reporter assay. Downregulation of CELF2 reversed the effects of inhibiting miR-20a expression.. Collectively, these results suggest a critical role for miR-20a in glioma cell apoptosis, proliferation, and invasion via the direct targeting of CELF2 and indicate its potential application in cancer therapy.

Topics: Apoptosis; Brain Neoplasms; CELF Proteins; Cell Line, Tumor; Cell Proliferation; Female; Flow Cytometry; Gene Expression Regulation, Neoplastic; Glioma; Humans; In Situ Nick-End Labeling; Male; MicroRNAs; Neoplasm Invasiveness; Nerve Tissue Proteins; RNA, Messenger; RNA, Small Interfering; Sincalide; Transfection

Glioma is a highly aggressive and lethal brain tumor. Signal transducers and activators of transcription (STAT) pathway are widely implicated in glioma carcinogenesis. Our previous study found that the Fynrelated kinase (FRK) gene, plays as a tumor suppressor in the development and progression of glioma. This study aimed to investigate the role of FRK in the activation pathway of STATs and its effect on the growth of glioma.. The U251 and U87 cells with stable FRK overexpression were generated by lentivirus technique. The effects of FRK on the related proteins of STAT signaling pathway were detected by western blotting. Coimmunoprecipitation was used to detect the association of FRK and STAT1. The effects of STAT1 on the proliferation of glioma cells were detected by CCK8 or Edu cell proliferation assays. The expressions and correlation of FRK and p-STAT1 in glioma tissues were detectd by western blotting or immunohistochemistry. The effect of FRK on the growth of glioma was investigated in vivo mouse model.. The level of p-JAK2 and p-STAT1 increased after FRK overexpression, while they decreased after FRK downregulation both in U251 and U87 cells. However, FRK had no effect on STAT3 phosphorylation. FRK-induced STAT1 activation was not dependent on JAK2. FRK associated with STAT1, induced STAT1 nuclear translocation and regulated the expressions of STAT1-related target genes. STAT1 overexpression suppressed the proliferation of glioma cells. In contrast, STAT1 knockdown by siRNA promoted glioma cell growth. Importantly, down-regulation of STAT1 partially attenuated FRK-induced growth suppression. The clinical sample-based study indicated that the expression of FRK was significantly correlated with the expression of p-STAT1. FRK significantly inhibited glioma tumor growth in vivo.. Our findings highlighted a critical role of FRK in tumor suppression ability through promoting STAT1 activation, and provided a potential therapeutic target for glioma.

Topics: Active Transport, Cell Nucleus; Animals; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Disease Progression; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Glioma; HEK293 Cells; Humans; Janus Kinase 2; Male; Mice, Nude; Neoplasm Proteins; Neoplasm Transplantation; Phosphorylation; Protein-Tyrosine Kinases; Signal Transduction; Sincalide; STAT1 Transcription Factor; Tumor Burden

Temozolomide (TMZ) is the preferred chemotherapeutic drug approved for the Glioblastoma multiforme (GBM) treatment. However, resistance to TMZ is the most intractable challenge for treatment of GBM. Screening of miRNAs is becoming a novel strategy to reveal underlying mechanism of drug-resistance of human tumors.. We conducted RNA sequencing (RNA-seq) for GBM cells treated continuously with TMZ 1 or 2 week or not. Bioinformatic analysis was used to predict targets of these altered miRNAs. Subsequently, we studied the potential role of miR-1268a in TMZ-resistance of GBM cells.. Expression levels of 55 miRNAs were identified altering both after 1 and 2 weeks TMZ treatment. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to illuminate the biological implication and related pathways of predicted target genes. We showed that miR-1268a was downregulated after TMZ treatment and targeted ABCC1/MRP1, a membrane transporter contributing to drug resistance, using dual-luciferase assay. Furthermore, we confirmed overexpression of miR-1268a inhibited protein translation of ABCC1 and restored upregulated expression of ABCC1 due to TMZ. Inversely, knockdown of miR-1268a increased ABCC1 at protein level and enhanced upregulation of ABCC1 with TMZ treatment. In addition, our data indicated that miR-1268a enhanced TMZ sensitivity in GBM cells.. Through RNA-seq analysis, we discovered miR-1268a and elucidated its role in modulating TMZ-resistance of GBM cells by targeting ABCC1.

Topics: Animals; Antineoplastic Agents, Alkylating; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Glioblastoma; Humans; Kaplan-Meier Estimate; Mice, Nude; Microarray Analysis; MicroRNAs; Multidrug Resistance-Associated Proteins; RNA, Messenger; Sincalide; Temozolomide; Time Factors; Transfection; Xenograft Model Antitumor Assays

Gliomas are highly malignant tumors, the most common of which are astrocytomas. A growing number of studies suggest that dysregulation of miRNAs is a frequent event contributing to the pathogenesis of gliomas. In this study, we found that over-expression of miR-132 inhibited cell proliferation and migration and triggered apoptosis, while knockdown of miR-132 showed opposite effects. PEA-15 was identified as a direct target of miR-132. Reintroduction of PEA-15 without 3'UTR region reversed the inhibitory effects of miR-132 on cell proliferation, migration, and apoptosis. MiR-132 was inversely correlated with the PEA-15 expression. CREB (cAMP response element binding protein) and KLF (Krüppel-like factor 8) were conformed as transcription factors of miR-132, which bidirectionally regulate the expression of miR-132. Our study suggests that miR-132 is an important tumor suppressor of astrocytoma progression by targeting PEA-15, while CREB and KLF can modulate the expression of miR-132, thus providing new insight into the molecular mechanisms underlying astrocytoma progression in vitro.

Topics: Analysis of Variance; Apoptosis; Apoptosis Regulatory Proteins; Astrocytoma; Brain Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; CREB-Binding Protein; Disease Progression; Gene Expression Regulation, Neoplastic; Glioma; HEK293 Cells; Humans; Intracellular Signaling Peptides and Proteins; Kruppel-Like Transcription Factors; MicroRNAs; Phosphoproteins; Repressor Proteins; RNA, Messenger; Sincalide; Time Factors; Transfection

Drosophila lethal (2) giant larvae (lgl) has been reported as a tumor suppressor and could regulate the Drosophila hippo signaling. Human giant larvae-1(Hugl-1), one human homologue of Drosophila lgl, also has been reported to be involved in the development of some human cancers. However, whether Hugl-1 is associated with the pathogenesis of malignant gliomas remains poorly understood. In the present work, we examined the effect of Hugl-1 on glioma cell growth both in vitro and in vivo. Firstly, we found that Hugl-1 protein levels decreased in the human glioma tissues, suggesting that Hugl-1 is involved in glioma progression. Unfortunately, either stably or transiently over-expressing Hugl-1 did not affect glioma cell proliferation in vitro. In addition, Hugl-1 over-expression did not regulate hippo signaling pathway. Interestingly, over-expression of Hugl-1 not only inhibited gliomagenesis but also markedly inhibited cell proliferation and promoted the apoptosis of U251 cells in an orthotopic model of nude mice. Taken together, this study provides the evidence that Hugl-1 inhibits glioma cell growth in intracranial model of nude mice, suggesting that Hugl-1 might be a potential tumor target for glioma therapy.

Topics: Animals; Astrocytoma; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Cytoskeletal Proteins; Disease Models, Animal; Female; Gene Expression Regulation, Neoplastic; Green Fluorescent Proteins; Humans; Ki-67 Antigen; Male; Mice; Mice, Nude; Phenylurea Compounds; Sincalide; Transfection

RBQ3, also known as RBBP5 (RB-binding protein 5), was an RB-binding protein. Besides, it was one of core components of MLL1 (mixed lineage leukemia 1), which were required for H3K4 methyltransferase activity. MLL1 dysfunction was found to be associated with the progression of some cancers such as acute leukemias. However, the precise role of RBQ3 in tumor progression remains obscure. In this study, we explored the expression and clinical role of RBQ3 in gliomas. Our results showed that RBQ3 was significantly upregulated in clinical glioma specimens by Western blot and immunohistochemistry. Moreover, its level was significantly associated with the pathology grades. High RBQ3 expression was suggested to be an independent prognostic factor for glioma patients' survival by univariate and multivariate analyses. Serum starvation and refeeding assay indicated that the expression of RBQ3 increased 8h after serum-stimulation, together with percentage of cells at S phase. In addition, knockdown of RBQ inhibited U87-MG cell proliferation with CCK8 kit, flow cytometry assays and colony formation analyses; while the depletion of RBQ3 induced the apoptosis of U87-MG cells. All the findings suggested that RBQ3 might play an important role in glioma, and RBQ3 inhibitors might be novel anti-tumor agents.

Topics: Adult; Apoptosis; Brain; Brain Neoplasms; Cell Line, Tumor; Chi-Square Distribution; Colony-Forming Units Assay; Culture Media, Serum-Free; DNA-Binding Proteins; Female; Gene Expression Regulation, Neoplastic; Glioma; Humans; Ki-67 Antigen; Male; Middle Aged; Nuclear Proteins; RNA, Small Interfering; Sincalide; Transfection

Studies have shown that several miRNAs play important roles in regulating a variety of cellular processes in gliomas. In these reports, upregulation of miR-193b has been found to be associated with a poor prognosis for glioma, but its functional mechanism in glioma remains unclear. This study investigates the roles of miR-193b in glioma tumor growth. We first showed that the expression of miR-193b was elevated in both glioma samples and glioma cells. Furthermore, downregulation of miR-193b by inhibitors was statistically correlated with a decrease in cell growth and a restored G1 accumulation. Luciferase assay and Western blot analysis revealed that Smad3 is a direct target of miR-193b. To prove that miR-193b regulated cell growth through the transforming growth factor-β (TGF-β) pathway in glioma cells by regulating Smad3, we tested endogenous targets of the TGF-β pathway by measuring the accumulation of p21 mRNAs after downregulation of miR-193b. The results confirmed that induction of p21 was promoted by miR-193b inhibitors in glioma cells, although this induction disappeared when Smad3 was knocked down with siRNA. Moreover, downregulation of Smad3 mitigates the miR-193b suppression of glioma proliferation. In conclusion, these results suggest that miR-193b regulated cell growth in glioma through the TGF-β pathway by regulating Smad3. Thus, our study indicates that miR-193b promotes cell proliferation by targeting Smad3 in human glioma, which may serve as a potentially useful target for development of miRNA-based therapies in the future.

Topics: Adult; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Glioma; HEK293 Cells; Humans; Male; MicroRNAs; Oligonucleotides; RNA, Small Interfering; Sincalide; Smad3 Protein; Transfection; Transforming Growth Factor beta; Tumor Cells, Cultured; Young Adult

MicroRNAs(miR) play an important role in cell growth, differentiation, proliferation and apoptosis, which can function either as oncogenes or as tumor suppressors in their effect on tumor growth. Smad3 is often underexpressed in very diverse types of malignant tumors and has an important tumor suppressive function; however, the underlying mechanism in solid cancer including glioblastomas(GBM) is not fully explored. The aim of this study is to explore the role of miR-92b in regulation of smad3 in GBM. In our study, we found that miR-92b expression was significantly increased in GBM tissues compared with normal brain tissues by Q-RT-PCR and in situ hybridization (P<0.01). However, expression of smad3 in GBM samples was significantly reduced compared with normal brain tissues by western blot and immunohistochemistry (P<0.05). Using 3'UTR luciferase reporter gene assay, we found that miR-92b directly affected smad3 expression in GBM cells by targeting the 3'-untranslated region. Silencing of miR-92b was able to significantly inhibit the viability of GBM cells in three GBM cell lines through up-regulating the TGF-beta/smad3/p21 signaling pathway in vitro. Furthermore, the tumor growth and the weight of U87 cells in the miR-92b inhibitor group were significantly inhibited when compared with that of the control group in vivo. Our data demonstrated that miR-92b may be considered as a tumor oncogene to promote GBM cell proliferation, and thus may serve as a potentially useful target for development of miRNA-based therapies in the future.

Topics: Animals; Apoptosis; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p21; Female; Gene Expression Regulation, Neoplastic; Glioblastoma; Humans; Luciferases; Mice; Mice, Nude; MicroRNAs; Oligonucleotides; RNA, Messenger; RNA, Small Interfering; Sincalide; Smad3 Protein; Time Factors; Transfection; Xenograft Model Antitumor Assays

The purposes of our study were to elucidate the role of BRG1 in the development of human glioma and to determine the effect of BRG1 on glioma cell growth, migration and invasion.. Using tissue microarray and immunohistochemistry, we evaluated BRG1 staining in 190 glioma tissues, 8 normal brain tissues and 8 tumor adjacent normal brain tissues. We studied glioma cell proliferative ability with reduced BRG1 expression by siRNA using CCK-8 cell proliferation assay and cell cycle analysis. We studied the role of BRG1 in glioma cell migration and invasion by cell migration assay and matrigel invasion assay. We performed western blot to detect cyclin D1, cyclin B1 and MMP-2 protein expression. We also detected MMP-2 enzyme activity by gelatin zymography.. Our results showed that BRG1 expression was increased in benign tumor and malignant tumor compared with tumor adjacent normal brain tissue (P < 0.01 for both). We did not find any correlation between BRG1 expression and clinicopathological parameters. In addition, we found that knockdown of BRG1 in glioma cell lines inhibits cell growth due to the G1 phase arrest by downregulating cyclin D1. We further demonstrated that silencing of BRG1 in glioma cells inhibited the cell migration and invasion abilities, and downregulation of MMP-2 expression greatly contributed to the reduced cell invasion and migration abilities.. Our data indicated that BRG1 expression is significantly increased in human glioma and it may be involved in the process of glioma cell proliferation, migration and invasion.

Topics: Brain Neoplasms; Cell Cycle; Cell Growth Processes; Cell Line, Tumor; Cell Movement; Cyclin B1; Cyclin D1; DNA Helicases; Female; G1 Phase Cell Cycle Checkpoints; Gene Knockdown Techniques; Gene Silencing; Glioma; Humans; Immunohistochemistry; Male; Matrix Metalloproteinase 2; Middle Aged; Neoplasm Invasiveness; Nuclear Proteins; RNA, Small Interfering; Sincalide; Transcription Factors

Human astrocytic tumors grow into the normal brain parenchyma either as localized tumors, or as highly diffuse neoplasms. The diffuse phenotype relates to a specific sub-type of neoplastic astrocytes with a high motility and invasion capacity. Motility features refer to locomotion while invasion features refer to protease secretion. Our data reveal that several peptides belonging to the gastrin/cholecystokinin peptide class are able to significantly (and in certain cases very significantly) modify the level of tumor growth (at the level of cell proliferation and/or cell death), of motility and of invasion in various experimental models of human astrocytic tumors. We are synthesizing various gastrin/cholecystokinin-related peptides in order to develop clinical applications with which we want to inhibit astrocytic tumor growth, individual neoplastic astrocytic motility and the invasion of the normal brain parenchyma.

Topics: Animals; Astrocytoma; Biomarkers, Tumor; Brain Neoplasms; Disease Models, Animal; Drug Evaluation, Preclinical; Gastrins; Humans; Mice; Sincalide

Gastrin and cholecystokinin (CCK) mediate their effects through at least two types of receptors (CCK receptors A and B). While it has been hypothesized that gastrin, a stimulator of gastric acid secretion, is also a neurotransmitter and a stimulator of cell proliferation in various normal and neoplastic tissues, its effect on astrocytic brain tumors has not been actively investigated.. Our goal was to determine the effects of gastrin and gastin and/or CCK antagonists on the proliferation in vitro of astrocytic tumor cells by use of both established cell lines and primary cell cultures of tumor tissue.. Ten established astrocytic tumor cell lines, SW1088, SW1783, Hs683, H4, U87, U118, U138, U373, T98G, and A172, were studied. The effects of added gastrin (at 0.01, 0.1, and microM) and the gastrin/CCK antagonists L-365,260, CI-988, L-364,718, and JMV 234 (each at 0.01, 0.1, and 1 microM) on the cellular proliferation rates of the 10 cell lines were indirectly measured by use of the colorimetric tetrazolium assay. The influence of gastrin (at 0.01 microM) on the cellular proliferation of primary cultures from nine freshly explanted astrocytic tumors was assessed by means of tritiated thymidine uptake and autoradiography.. At specific concentrations, added gastrin increased the cellular proliferation of three established astrocytic cell lines (A172, Hs683, and SW1088), decreased it in two (U373 and T98G), and was without effect on the remaining five. Gastrin decreased cellular proliferation in one primary astrocytic tumor cell culture, stimulated it in five, and had no apparent effect in the remaining three. L-365,260, a CCK receptor B antagonist used at 0.01 microM, increased cellular proliferation in seven cell lines (A172, H4, Hs683, SW1783, T98G, U118, and U138), decreased it in one (U87), and had no effect in the remaining two. CI-988, another CCK receptor B antagonist used at 0.01 microM, inhibited cellular proliferation in five cell lines (A172, H4, SW1783, U373, and U87), stimulated it in two (T98G and U138), and had no effect in three. The CCK receptor A antagonists L-364,718 and JMV 234, both used at 0.01 microM, affected the cellular proliferation of only three of the 10 cell lines.. These results suggest that gastrin (and perhaps CCK that belongs to the same peptide family) may play a role in the growth of a substantial proportion of human astrocytic tumors.

Topics: Amino Acid Sequence; Astrocytoma; Autoradiography; Benzodiazepinones; Brain Neoplasms; Cell Division; Cholecystokinin; Devazepide; Gastrins; Hormone Antagonists; Humans; Indoles; Meglumine; Molecular Sequence Data; Peptide Fragments; Phenylurea Compounds; Receptors, Cholecystokinin; Sincalide; Thymidine; Tritium; Tumor Cells, Cultured

We have investigated the potential role of gastrin in the regulation of cell growth in human astrocytic tumors. To this end we have used synthetic analogs of gastrin and cholecystokinin (CCK) which behave as gastrin and/or CCK antagonists, e.g. compounds JMV-97, JMV-209 and JMV-179. Their effects on astrocytic tumor cell proliferation was investigated by the use of the colorimetric MTT assay. The in vitro biological models used in the present study included the SW1088, U87 and U373 astrocytic tumor cell lines. The results demonstrated marked influence of gastrin and CCK antagonists in the regulation of astrocytic tumor growth. This suggests that gastrin and/or CCK antagonists might be tested in experimental glioblastoma.

Topics: Astrocytes; Astrocytoma; Brain Neoplasms; Cholecystokinin; Gastrins; Glioblastoma; Humans; Peptide Fragments; Sincalide; Tumor Cells, Cultured