The Role Of CDKN2A Gene In High-Grade Glioma And Its Relationship With Drug Resistance

Objective: To explore the role of CDKN2 A gene in the pathogenesis of High-Grade glioma.Methods: i TRAQ technology was used to investigate the differential expression of proteins in tumor tissues of patients with high grade gliomas compared with non-tumor patients.The target gene of this study was determined according to the enrichment of go function and the enrichment of KEGG pathway;the enrichment of go was mainly the metabolism process of nucleic acid,and the enrichment of KEGG pathway was mainly the metabolism pathway containing nucleotide;then according to the results of i TRAQ,cas9 base was used CDKN2 A gene was targeted by knockout system to establish knockout cell line,and CDKN2 A gene stable expression cell line was established by lentivirus combined with drug screening.WST-1 was used to draw the cell proliferation activity of knockout group,untreated group and stable expression group;flow cytometry was used to detect the cell cycle of knockout group,untreated group and stable expression group;ELISA was used to detect the content of Cyclin D1 and p Rb protein in knockout group,untreated group and stable expression group;scratch test was used to detect the content of Cyclin D1 and p Rb protein in knockout group,untreated group and stable expression group after adding the same drugs 24-hour closure rate.Results:(1)1074 differential proteins were found in i TRAQ,616 of which were up regulated and 458 of which were down regulated.The enrichment of differential protein go and KEGG mainly focused on the synthesis and metabolism of nucleic acid;(2)with the increase of cell number,the growth rate of knockout group was higher than that of untreated group and stable expression group,and the difference was statistically significant.The growth rate of stable expression group was lower than that of knockout group and untreated group,and the difference was statisticallysignificant.(3)The cell cycle of knockout group was longer than that of stable expression group in G0-G1 phase,which showed that CDKN2 A gene mainly inhibited cell growth from G0-G1 phase.(4)The protein contents of cyclin D1 and p RB in U251 and U87 cells which stably expressed CDKN2 A gene were lower than those in knockout group and untreated group.The knockout group had the highest content,and the difference was statistically significant(P< 0.05).(4)The closure rate of CDKN2 A gene in U251 cell lines decreased 24 hours after overexpression.Conclusion:(1)There is abnormal expression of CDKN2 A gene in glioma cells.CDKN2 A gene is involved in the occurrence and development of glioma.(2)CDKN2A stable overexpression can effectively inhibit the proliferation of glioma cells.If CDKN2 A gene expression is inhibited,glioma cells will increase.(3)CDKN2A gene mainly acts on cell cycle.The stable overexpression of CDKN2 A gene reduces the number of cells in go phase,indicating that the gene has the effect of stagnating cell cycle in go,and its action pathway may be CDKN2A-Cyclin D1-PRB pathway.If the pathway is inactivated or mutated,it may cause cell proliferation to accelerate and apoptosis to be blocked.CDKN2 A gene plays an important role in the occurrence and development of glioma,which may be a potential therapeutic target for glioma.(4)When CDKN2 A gene is over expressed,glioma cells are more sensitive to the antitumor effect of carmustine.CDKN2 A gene may be a potential site to overcome the drug resistance of glioma.