Na Ban

Nucleostemin promotes the proliferation of human glioma via Wnt/β-Catenin pathway

Nucleostemin, nucleolar guanosine triphosphate (GTP)‐binding protein 3, is a member of the MMR1/HSR1 GTP‐binding protein family. The important roles of nucleostemin in self‐renewal, cell cycle regulation, apoptosis, and cell proliferation of various cancer types as been shown. Nevertheless, its expression and potential functions in human glioma is still unclear. In the present study, we demonstrated that up‐regulation of nucleostemin was tightly related to poor 5‐year‐survival ratios. In serum‐starved and re‐feeding models of U251 and U373MG, we observed the rising expression of nucleostemin and p‐β‐Catenin (p‐Tyr645) were accompanied with cell proliferation markers (cyclin D1 and proliferating cell nuclear antigen (PCNA)). Employing nucleostemin‐depletion models, we found down‐regulated nucleostemin and p‐β‐Catenin. The flow cytometry analysis proved the weakened cell proliferation. Moreover, we detected the translocation of β‐Catenin into the nucleus was impaired, meaning the inhibition of the Wnt/β‐Catenin pathway. Taken together, we identified a positive correlation between up‐regulation of nucleostemin and human glioma cell proliferation and that knocking‐down nucleostemin alleviated glioma proliferation by reducing β‐Catenin transportation into the nucleus. All results suggested that nucleostemin might accelerate human glioma proliferation via the Wnt/β‐Catenin pathway.

Knockdown of PFTK1 Inhibits the Migration of Glioma Cells

The prognosis of glioma patients is generally poor, so it is urgent to find out the underlying molecular mechanisms. PFTK1 is a member of cyclin-dependent kinases (Cdks) family and has been reported to contribute to tumor migration and invasion. In this study, we aimed to explore the expression and function in human glioma. Western blot and immunohistochemistry were used to evaluate the expression of PFTK1. PFTK1 expression was higher in glioma tissues compared with normal brain tissues, and its level was associated with the WHO grade in Western blot analysis. The suppression of PFTK1 expression by RNA interference was shown to inhibit the migration of glioma cells. Knockdown of PFTK1 increases E-cadherin expression and decreases vimentin expression. These data show that PFTK1 may participate in the pathogenic process of glioma, suggesting that PFTK1 can become a potential therapeutic strategy for gastric cancer.

Overexpression of CCT8 and its significance for tumor cell proliferation, migration and invasion in glioma

Overexpression of chaperonin containing t-complex polypeptide 1 (TCP1), or CCT, has been reported in various classes of malignancies. However, little is known about the expression of t-complex protein subunits TCP1theta (CCT8) in gliomas. In this study, the expression of CCT8 protein was detected using blotting analysis and immunohistochemistry. CCT8 was found to be overexpressed in gliomas and to correlate with the WHO grade of gliomas. To further investigate the biological function of CCT8 in gliomas, CCT8-silenced U87 glioblastoma multiforme (GBM) and U251MG cells were constructed using a small interference RNA (siRNA) sequence. The knockdown effect of CCT8 on proliferation and invasion in these cells was analyzed using the CCK8, flow cytometry cycle, scratch, transwell invasion and fluorescence assays. Compared with the controls, the glioma cells expressing CCT8-siRNA exhibited a significantly decreased proliferation and invasion capacity, as well as a dysregulated cell cytoskeleton. This study showed that high CCT8 protein expression might be related to poor outcome of glioma, and that CCT8 regulates the proliferation and invasion of glioblastomas.

The role of FoxJ2 in the migration of human glioma cells.

Previous studies have demonstrated that FoxJ2 (forkhead box J2) is a member of Forkhead Box transcription factors and acts as an important prognostic indicator in human breast cancer. Our study aimed to assess the expression and function in human glioma. Western blot analysis and immunohistochemistry were performed in human glioma tissues. Low FoxJ2 expression was observed in 80 samples and its level was correlated with the grade of malignancy. A strongly positive correlation was observed between FoxJ2 and E-cadherin. Overexpression of FoxJ2 increased E-cadherin expression and decreased vimentin expression. The wound healing and transwell assays showed that overexpression of FoxJ2 significantly inhibited their migration in U87 cells. Consistent with this, knockdown of FoxJ2 promoted cellular motility. In a word, FoxJ2 suppressed cell migration and invasion in glioma, which might be a potential novel molecular targeted therapy for surgery and immune treatment.

Expression of CDC5L is associated with tumor progression in gliomas

Cell division cycle 5-like (CDC5L) protein is a cell cycle regulator of the G2/M transition and has been reported to participate in the catalytic step of pre-messenger RNA (mRNA) splicing and DNA damage repair. Recently, it was also found to act as a candidate oncogene in osteosarcoma and cervical tumors. However, the role of CDC5L expression in tumor biology was still unclear. Here, we analyzed the expression and clinical significance of CDC5L in gliomas. The expression of CDC5L in fresh glioma tissues and paraffin-embedded slices was evaluated by western blot and immunohistochemistry, respectively. We found that CDC5L was highly expressed in glioma tissues. The expression of CDC5L was significantly associated with glioma pathology grade and Ki-67 expression. Univariate and multivariate analyses showed that high CDC5L expression was an independent prognostic factor for glioma patients’ survival. To determine whether CDC5L could regulate the proliferation of glioma cells, we transfected glioma cells with interfering RNA target CDC5L, then investigated cell proliferation with cell counting kit (CCK)-8, flow cytometry assays and colony formation analyses. Our results indicated that knockdown of CDC5L would inhibit proliferation of glioma cells. Besides, reduced expression of CDC5L could induce the apoptosis of glioma cells. These findings suggested that CDC5L might play an important role in glioma and thus be a promising therapeutic target of glioma.

LAP3 promotes glioma progression by regulating proliferation, migration and invasion of glioma cells

Leucine aminopeptidase 3 (LAP3), belonging to the M1 family, has been proved to catalyze the hydrolysis of leucine residues. Leucine aminopeptidases are involved in many pathological disorders and regulate cell proliferation, invasion and/or angiogenesis of tumor. Recent study showed that LAP3 is highly expressed in several malignant and affects tumor angiogenesis. We aimed at investigating the clinical significance of LAP3 expression in human gliomas and its biological function in glioma cells. RT-PCR, Western blot and immunohistochemistry analysis were used to detect the expression levels of LAP3 in 121 glioma tissues, high LAP3 expression was correlated with the grade of malignancy and poor prognosis of glioma patients. In vitro, after increasing of LAP3 by myc-LAP3 transfection and knockdown of LAP3 by siRNA transfection in glioma cells, cell viability, cell cycle, migration and invasion were determined with CCK8 assay, flow cytometry, wound healing and transwell invasion assays. The results indicated that increasing LAP3 could promte cell viability, cell cycle, migration and invasion, knockdown LAP3 could decrease cell viability, suppress cell proliferation, migration and invasion. Our findings uncover that LAP3 might be a new prognostic factor and be close correlation with glioma cell growth, migration, invasion.

High expression of adenylate cyclase-associated protein 1 accelerates the proliferation, migration and invasion of neural glioma cells

Adenylate cyclase-associated protein 1 (CAP1), a conserved member of cyclase-associated proteins was reported to be associated with the proliferation, migration or invasion of the tumors of pancreas, breast and liver, and was involved in astrocyte proliferation after acute Traumatic Brain Injury (TBI). In this study, we sought to investigate the character of CAP1 in the pathological process of human glioma by detecting human glioma specimens and cell lines. 43 of 100 specimens showed high expression of CAP1 via immunohistochemistry. With statistics analysis, we found out the expression level of CAP1 was correlated with the WHO grades of human glioma and was great positively related to Ki-67 (p<0.01). In vitro, silencing CAP1 in U251 and U87MG, the glioma cell lines with the relatively higher expression of CAP1, induced the proliferation of the cells significantly retarded, migration and invasion as well. Obviously, our results indicated that CAP1 participated in the molecular pathological process of glioma indeed, and in a certain sense, CAP1 might be a potential and promising molecular target for glioma diagnosis and therapies in the future.

CHD1L Regulates Cell Cycle, Apoptosis, and Migration in Glioma

Chromodomain helicase/ATPase DNA binding protein 1-like (CHD1L) gene is a newly identified oncogene located at Chr1q21 and it is amplified in many solid tumors. In this study, we intended to investigate the clinical significance of CHD1L expression in human glioma and its biological function in glioma cells. Western blot and immunohistochemistry analysis showed that CHD1L was overexpressed in glioma tissues and glioma cell lines. In addition, the expression level of CHD1L was positively correlated with glioma pathological grade and Ki-67 expression. Kaplan–Meier curve indicated that high expression of CHD1L may result in poor prognosis of glioma patients. Accordingly, suppression of CHD1L in glioma cells was shown to induce cell cycle arrest and increase apoptosis. In addition, knockdown of CHD1L significantly accelerated migration and invasion ability of glioma cells. Together our findings suggest that CHD1L is involved in the progression of glioma and may be a novel target for further therapy.

DTX3L is upregulated in glioma and is associated with glioma progression

Gliomas are the most common primary brain tumors of the central nervous system (CNS). Due to the poor prognosis of glioma patients, it is urgent to develop more effective therapies. Deltex-3-like (DTX3L), also known as B-lymphoma and BAL-associated protein (BBAP), has been reported to play an important role in the progression of many tumors. This study aimed to investigate the clinical significance and biological function of DTX3L in human glioma. Clinically, the protein expression level of DTX3L is increased in glioma tissues compared with that observed in normal brain tissues. Immunohistochemical analysis demonstrated that DTX3L was highly expressed in the glioma tissues and its level was correlated with the grade of malignancy. Multivariate analysis revealed the association between high expression of DTX3L and the poor prognosis of glioma patients. In addition, knockdown of DTX3L by siRNA transfection increased glioma cell apoptosis. Moreover, suppression of DTX3L expression was shown to significantly inhibit the migration and invasion of glioma cells. These data indicate that DTX3L plays an important role in the pathogenic process of glioma, suggesting that DTX3L could be a potential prognostic biomarker for glioma.

The role of Alix in the proliferation of human glioma cells

Apoptosis-linked-gene-2-interacting protein 1 (Alix) is involved in the endosome-lysosome system in the cytoplasm. The normal function of Alix may be altered by ALG-2 toward a destructive role during active cell death. Alix also may play a role in regulation of cell proliferation. However, the role of Alix in human glioma has not been elucidated yet. This study intended to clarify the relationship between Alix and glioma pathologic grades and its role in the proliferation of glioma cells. Our findings showed that Alix protein concentrations were significantly elevated in high-grade glioma tissue compared with low-grade glioma (P < .0001). Immunohistochemical study revealed that Alix was overexpressed in 75 resected glioma tissues and may forecast poor survival. Alix expression was increased in resting serum-stimulated glioma cells. Additionally, we reduced Alix expression in U251MG cells and then found that cell viability was decreased significantly when p21 expression increased. Colony formation assay and flow cytometry analysis demonstrated that reduced Alix expression may lead to growth inhibition and cell cycle arrest. In summary, our findings suggest that Alix plays an important role in the proliferation of glioma cells and may be a novel therapeutic target.