The Expression Of DKC1 In Glioma And Its Effect On Proliferation,Migration And Invasion Of Glioma Cells And Its Mechanism

Objective Glioma is still one of the most difficult problems in neurosurgery because of its high invasiveness in brain tissue.DKC1 is currently considered to be an oncogene,which is highly expressed in many types of tumors,It is closely related to the prognosis of patients,and its molecular mechanism in tumorigenesis and development is also actively explored.This study will initially explore the expression of DKC1 gene in human glioma and its correlation with the degree of malignancy of glioma.And further study the effect of knockdown of DKC1 gene expression on the proliferation,migration and invasion of glioma U251 and U87 cells and its related mechanisms,providing a theoretical basis for the molecular mechanism of glioma and the exploration of new therapeutic targets.Methods Tissue microarray combined with immunohistochemical was used to observe the expression of DKC1 in glioma tissues.A total of 70 glioma tissue specimens were included in the study,34 cases of low-grade(WHOI-II)human glioma tissues and 36 cases of high-grade(WHOIII-IV)glioma tissues,8 cases of normal control(taken from traumatic decompression brain tissue),The relationship between DKC1 and glioma malignancy and its significance in prognosis were explored.The glioma cell lines U251 and U87 cells were selected according to the expression of DKC1 and divided into three groups: negative control group(si-Con),small interference group1(si-1),and small interference group2(si-2),respectively,through si Lent Fect Negative control si RNA,DKC1 si RNA-1,DKC1 si RNA-2 were transfected into glioma cells;Western blot was used to detect the expression of DKC1 protein in glioma cells before and after transfection,The proliferation ability of glioma cells in the control group and the small interference group was detected by the cell proliferation assay and cell cycle experiment;The migration ability of glioma cells in the control group and the small interference group was detected by cell scratching test and Transwell cell migration assay;The invasion ability of glioma cells was detected by transwell cell invasion assay.The effect of DKC1 expression on Cyclin E2,CDK2 and P27 was detected by Western blot to investigate the mechanism of DKC1 on glioma cell proliferation.And the effect of DKC1 expression on migration-related proteins in N-cadherin,MMP2 and HIF-1α cells was also detected by Western blot to investigate the mechanism of DKC1 on migration and invasion of glioma cells.Results 1.DKC1 expression level in glioma tissues was significantly higher than that in normal brain tissues(p<0.05),and was significantly positively correlated with WHO pathological grade(p<0.01).The expression level of DKC1 in high-grade(WHOIII-IV)glioma was significantly higher than that in the low-grade(WHOI-II)glioma group.However,there was no significant difference in DKC1 expression between glioma tissues of different ages and genders(p>0.05).The Kaplan-Meier survival curve showed that the survival time of DKC1 high expression group was significantly lower in the high-grade glioma patients than in the low-expression group,suggesting that DKC1 expression is closely related to the clinical prognosis of glioma patients.2 In the glioma U251 and U87 cell lines,the relative expression of DKC1 protein in the small interference group(si-1,si-2)was significantly lower than that in the si-Con group,the difference was statistically significant(p<0.01),suggesting that small interfering RNA successfully knocked down DKC1 expression.After using small interfering RNA to knock down DKC1 expression,the proliferation,migration and invasion ability of glioma cells were significantly lower than those of the control group,and the difference was statistically significant(p<0.05).3 The results of Western blot showed that the expression of Cyclin E2 and CDK2 in the small interference group was significantly decreased and the expression of P27 was up-regulated in the small interference group,the difference was statistically significant(p<0.05).The expression levels of N-cadherin,MMP2 and HIF-1α were also significantly decreased,and the difference was statistically significant(P<0.05).Conclusion 1.DKC1 is highly expressed in glioma tissues and glioma cells,and its expression level is positively correlated with glioma WHO pathological grade,and is closely related to the clinical prognosis of high-grade glioma patients.The survival time of DKC1 high expression group was significantly shorter than that of low expression group,and DKC1 could be used as an independent prognostic factor for glioma.2 Down-regulation of DKC1 can effectively inhibit the proliferation,migration and invasion of glioma U251 and U87 cells.3 Down-regulation of DKC1 may inhibit the proliferation of glioma cells by regulating the expression of Cyclin-E2,CDK2,and P27 proteins.4 Down-regulation of DKC1 may inhibit the migration and invasion of glioma cells by regulating the expression of N-cadherin,MMP-2 and HIF-1α proteins.