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Objective To explore the expression of the CENP-W in gliomas and investigate the effects of its invasion. Methods The expression level of the CENP-W in gliomas with varied pathologic grade were detected by immunohistochemical analysis,RealTime PCR,and Western Blotting. U251 cells were transfected with the specific siRNA to repress the CENP-W expression level. The invasion ability of U251 cells were examined by Transwell Chamber assay ,while RAS mRNA and protein levels were detected at the same time. Results The expression levels of the CENP-W in glioma tissues were significantly high and the CENP-W gene could enhance the invasion of U251 cells . The expression of RAS was down-regulated when the expression of CENP-W was repressed. Conclusion The CENP-W has an oncogenic role in human brain gliomas and may regulate the invasion of gliomas by adjusting the RAS signaling pathways.
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AIM:To study the effect of centromere protein W ( CENP-W) down-regulation on human glioma U87 cells.METHODS:Small interfering RNA ( siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot .The proliferation of the cells was analyzed by MTT assay , BrdU staining and colony formation experiment .Transwell chamber assay was used to detect the invasion a-bility of the cells .The cell migration ability was measured by a scratch test .The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry .RESULTS:The results of MTT assay , colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group .The results of Transwell and scratch experiments showed that the migra-tion and invasion abilities in experimental group were weaker than those in blank control group and negative control group . The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G 0/G1 phase .The percentage of apoptotic cells in experimental group was higher than that in control group ( P<0.05 ) .CON-CLUSION:Down-regulation of CENP-W expression inhibits the proliferation , migration and invasion of human glioma cells and promotes the apoptosis of the cells , suggesting that CENP-W may be a potential target of gene therapy for human glioma.
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Objective To investigate the expression of CENP-W in gliomas and its relationship with clinicopathological parameters and prognosis and to explore the effects of centromere protein W (CENP-W)on the invasion of gliomas cells.Methods The expressions of CENP-W in high-grade glioma tissues,low-grade glioma tissues,and adjacent brain tissues were detected by real-time fluorescence quantitative PCR and Western blotting. The correlation of the expression of CENP-W with clinicopathological parameters and prognosis was analyzed statistically.Human gliomas U2 5 1 cells in vitro were transfected with small interfering RNA to downregulate the expression of CENP-W.The invasion and migration capabilities of gliomas cancer cells were assessed by Transwell assays.Results The expression level of CENP-W was significantly higher in glioma tissues than in normal tissues. There was a positive correlation between the three protein expression levels and the pathological grade of gliomas. CENP-W siRNA was successfully transfected into U2 5 1 cells.Compared with those of the cells transfected with the scramble siRNA and control cells,the invasive and migration activities were inhibited in the U2 5 1 cells transfected with CENP-W siRNA.The Kaplan-Meier analysis and Log-Rank test showed significant differences in progress free survival (PFS)between the CENP-W high-expression and low-expression groups.Conclusion The expression level of CENP-W was positively correlated with the pathological grade of gliomas and CENP-W can promote glioma cell invasion.It implicates that CENP-W can be a novel target in gliomas treatment.
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Objective To explore the expressions and correlation of RWD containing sumoylation enhancer (RSUME),small ubiquitin-like modifier (SUMO-1),hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor (VEGF)in gliomas of different pathologic grade.Methods We investigated the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA with reverse transcription-polymerase chain reaction (RT-PCR),and investigated the immunohistochemical staining to determine the expressions of SUMO-1,HIF-1α and VEGF in 63 cases of human gliomas of different pathologic grade and 9 cases of normal brain tissues.We studied its correlation with the pathologic grade and the relationship between the expression of RSUME promoter sumoylation and HIF-1α/VEGF pathway in gliomas.Results There were significant differences (P <0.01)in the expressions of RSUME mRNA,HIF-1αmRNA and VEGF mRNA in glioma tissues.With the increasing degree of pathologic grade in tumor specimens,the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA increased markedly (P <0.01 ).There was a positive correlation of the expression levels of RSUME mRNA with HIF-1αmRNA and VEGF mRNA.There were significant differences (P <0.01 )in the expressions of SUMO-1,HIF-1αand VEGF in glioma tissues by immunohistochemical staining.With the ascending of pathologic grade of tumor specimens,the expression levels of SUMO-1,HIF-1α and VEGF increased markedly (P < 0.01 ).There was a positive correlation between the expression level of SUMO-1 and HIF-1α(r =0.857,P <0.01).The Kaplan-Meier analysis and Log-rank test showed significant differences in progress free survival (PFS)between the RSUME high-expression and low-expression groups (χ2 =36.032,P <0.01).Conclusion RSUME may enhance HIF-1α/VEGF pathway through sumoylation in gliomas.It implicates that RSUME is related to angiogenesis in gliomas and can promote tumor invasion and progression,indicating that RSUME can be a novel target in gliomas treatment.