Expression And Significance Of Centromere Protein W On Glioma

Objective:Gliomas are the most common malignant brain tumors, which is a group ofnerve cells in the tumor epithelial function can be characterized by a general term toglial cell differentiation, the incidence rate of all intracranial tumors nearly50%-60%.In recent years, despite the comprehensive application of various treatments, clinicalefficacy achieved some progress, but the survival time is short and quality of life isnot still high, most patients will eventually died of tumor recurrence and metastasis.Existing studies show that overexpression of oncogenic genes closely related to tumorsuppressor inactivation, and how the molecular mechanisms of cancer is unknown.CENP-W was first reported in2007, it belongs to CCAN (constitutivecentromere associated network) family, initially found to result in many tumor tissuesand mouse fibroblast cells with high tumorigenic as a putative oncogene therefore benamed CUG2(Cancer-upregulated gene2); while CENP-W plays an important rolein the central nervous system development and function of neurons involved in themaintenance of CENP-W knockout zebrafish embryos may lead to neurodegeneration.domestic and foreign CENP-W brain gliomas. In domestic and international researchdid not investigate the relationship between the expression of CENP-W and survivalprognosis in glioma.This study was to preliminary discussed the expression of CENP-W in glioma,the relationship of the expression and pathological grade (WHO classification)，whileanalyzed the relationship of glioma survival prognosis and CENP-W expression。Forfurther study of CENP-W，which may be involved in regulating the process ofgliomas occurrence and development, which can provided a new direction for thefurther treatment of glioma.Methods: The experimental design is divided into three steps1、 the first step by Real-Time PCR、Western blotting、immunohistochemistrydetected the CENP-W mRNA and protein expression differences control group ofnormal brain tissue (non-tumor brain tissue), the experimental group was four different pathological grade gliomas；data collected and applied statistics methods foranalysis.2、The second step was detected to the comparison of CENP-W and GAPDHexpression in A172, LN308, U118, U87, U251, LN229, SNB19, TJ899, TJ905gliomacell lines by Western blotting; data collected and applied statistics methods foranalysis.3、The third step：postoperative follow-up data were collected to statisticalanalysis the relation of all the relevant factors, which collected from the patient oftumor specimens, and to analyze the relationship between patient survival and theexpression of CENP-W in tissue.Results:1. RT-PCR melting curve analysis to be able to CENP-Α and β-actin were only asingle peak, with satisfactory results. mRNA expression levels: normal tissue （1.0±0.19）, level I （3.3±0.31）, level II (5.4±0.43), level III (3.5±0.42), level IV (8.2±0.49). CENP-W have different between pathological levels in glioma tissues andnormal brain tissue, there are significant differences in contrast with statisticallysignificant. CENP-W mRNA expression in high level of glioma was significantlyhigher than I, II, III.2. The relative expression of CENP-W protein tissue results: normal (0.46±0.062), level I (0.66±0.099), level II (0.85±0.069), level III (0.70±0.05), levelIV (1.02±0.058), The relative expression of the protein in glioma is relatively high.The relative expression of the protein in histological grade IV was normal controlgroup two times.3. Total protein was extracted from the cell lines to detect the expressing ofCENP-W protein by Western blotting，which compared to GAPDH, A172（0.89±0.11）、U87（0.71±0.08）、U251（0.71±0.07）、TJ899（0.83±0.09）、TJ905（1.08±0.11）compared with normal nerve cell group (0.24±0.08) relative proteinexpression was significantly increased, LN308(0.46±0.07), LN229(0.39±0.09) issoftly different, The relative expression of the protein is small in U118(0.21±0.07),SNB19(0.22±0.12) compared to normal nerve cells. Use grayscale software toscan, P <0.05considered statistically significant. It showed a medium-high expression in majority of glioma cell lines.4. Expression of CENP-W in human gliomas by immunohistochemistry methodfurther validated. The results showed that CENP-W has in medium-high expression,the nucleus was positive; whereas normal brain tissue without color. To analyzed therelationship between CENP-W and Two groups of gliomas using the chi-square test,P=0.014<0.05, and it have statistically meaningful differences between the twogroups. Statistical results are shown the positive of CENP-W rate of74.0%in glioma,which expression in highly malignant group accounted for81.4%,18.6%of lowmalignant group.5. Statistical analysis of prognostic factors in patients with glioma, from themultivariate analysis, the values of Grade and CENP-W were0.00and0.005.Therefore Grade and CENP-W correlated to survival prognosis in patients withglioma, others’ in patients with glioma have ittle effect. The high malignant group ispoor prognosis compare with the low malignant group. Patients with glioma survivelonger which CENP-W expression is negative（HR=0.325，P=0.005）, the positive issurvive short.Conclusions:1. CENP-W is upregulated in glioma and it is oncogenes.2. The expression of CENP-W mRNA, protein have a trend of increase withPathological level of glioma.3.The positive expression of CENP-W in glioma tissue by immunohistochemicalprompt for malignant gliomas which have a certain significance.