Research Of MORC2 Expression And Effect On Human Glioma

Part ? Investigate the MORC2 function in glioma with Bioinformatics analysisObjective:To investigate the MORC2 function in gliomaMethods:T test were used to analysis the MORC2 expression level in glioma data set GSEA4290, to compare the MORC2 expression difference between glioma and normal tissue. Gene set enrichment analysis (GSEA) were used to explore the gene set enriched by MORC2 in glioma.Results:In sample GSEA4290, in contrast to normal brain tissue (23 cases), MORC2 expression is significantly increased in gliomas (157 cases), P<0.05. GSEA indicate the high expression of MORC2 enriches the gene set of cell cycle, mitotic, DNA repair, DNA damage response. Their enrichment scores are 0.660,0.739,0.565,0.546, respectively, P<0.01.Conclusion:The expression of MORC2 is increased in glioma than normal brain tissue. The high expression of MORC2 is involved in cell cycle regulation, mitogenic, enhance DNA damage response and repair.Part II Expression of MORC2 on glioma tissuesObjective:To investigate the expression in human glioma tissues and its significance.Methods:Collect glioma and normal brain tissues in clinic. Use immunohistochemistry to detect the expression level of MORC2 in low-grade glioma, glioblastoma and normal brain tissues. The glioma patients were divided into MORC2 low expression group and MORC2 high expression group, to compare the prognostic difference.Results:It didn't show MORC2 high expression in all 10 cases of normal brain tissue. It showed MORC2 high expression in 7 cases of WHO ??? grade glioma tissue of all 24 cases, and MORC2 high expression in 15 cases of WHO ? grade glioma tissue of all 24 cases, and MORC2 high expression in 20 cases of WHO IV grade glioma tissue of all 22 cases. The positive expression rates were 0%?29.2%?62.5%?90.9%, respectively. Survival analysis showed that, MORC2 low expression group were significantly longer than MORC2 high expression group on progression-free survival and overall survival.Conclusion:MORC2 expression was significantly increased in glioma., and MORC2 expression is increased combined with the increase of glioma pathological grade.Part ? The effects of MORC2 on glioma proliferation and apoptosis in vitroObjective:To investigate the effects of MORC2 downregulation on cell proliferation and apoptosis in human glioma cell line U251.Methods:To design two kinds of MORC2-shRNA to transfected into U251 cells, the group that transfected with empty vector shRNA was control group. RT-PCR and Western blot was used to detect the interference efficiency, CCK-8 kit assay was used to detect the cell growth. Cell cycle was analyzed by flow cytometry, Annexin PE double staining was used to detect the apoptosis.Results:RT-PCR and Western blot showed MORC2 expression is significantly decreased on shRNAl group and shRNA2 group. CCK-8 kit assay showed that cell proliferation on shRNAl group and shRNA2 group was significantly lower than the control group; In the analysis of cell cycle by flow cytometry, we found the proportion of G2/M phase is decreased on shRNAl group and shRNA2 group, compared to the control group. Annexin PE staining results showed significant increase in early apoptosis on shRNA1 group and shRNA2 group than the control group.Conclusion:successfully using siRNA silencing technique to inhibit the expression MTORC2 in glioma cells, downregulation of MORC2 can inhibit the proliferation of glioma cells and promote early apoptosis of glioma cells, and make G2/M phase cells proportion decreased.Part ? The effects of MORC2 on glioma invasion and migration in vitroObjective:To investigate the effects of MORC2 downregulation on cell invasion and migration in human glioma cell line U251.Methods:Design two kinds of MORC2-shRNA to transfected into U251 cells, the group that transfected with empty vector shRNA was control group. Soft agar colony formation assay was used to detect the single cell proliferation ability. Transwell assay was used to detect the ability of cell invasion and migration.Results:Soft agar colony formation assay showed that the colony-forming number of shRNA1 group and shRNA2 group was significantly lower than the control group. Transwell assay showed that compared with the control group, the number of cells that across the membrane was significantly reduced on shRNAl group and shRNA2 group.Conclusion:MORC2 downregulation inhibits the single cell proliferation ability and the ability of cell invasion and migration on glioma.