CDKL2甲基化调控恶性胶质瘤细胞生长的作用机制研究

Study on the Mechanism of CDKL2 Methylation in Regulating the Growth of Malignant Glioma Cells

目的:探讨细胞周期蛋白依赖性激酶2(CDKL2)在脑胶质瘤中的表达情况和甲基化状态,并探索CDKL2甲基化对于恶性胶质瘤细胞生长的影响。方法:通过甲基化敏感限制酶偶联实时荧光定量PCR(MSRE-qPCR)分析正常组织和肿瘤组织中CDKL2的甲基化水平;实时荧光定量PCR检测CDKL2在正常组织和肿瘤组织的表达差异,评估CDKL2甲基化和mRNA表达水平的相关性;体外实验和体内实验分别验证CDKL2甲基化对于恶性胶质瘤细胞增殖迁移的影响并且通过甲基化抑制剂5-aza-2′-deoxycytidine进行检测。结果:受试者工作特征曲线(ROC)分析显示,非肿瘤组织和胶质瘤组织的CDKL2的甲基化具有较高的特异度和敏感度(P<0.001)。相较于正常组织,CDKL2在胶质瘤组织中呈现高甲基化而低表达(P<0.001),CDKL2的甲基化水平和表达水平成反比。5-aza-2′-deoxycytidine可以有效抑制肿瘤的形成和肿瘤细胞的生长(P<0.01)。结论:胶质瘤细胞CDKL2的甲基化水平高,去甲基化可以抑制肿瘤细胞的生长,可能成为治疗胶质瘤的潜在靶点。

Objective:To explore the effect of CDKL2 methylation on the growth of malignant glioma cells.Methods:Methylation levels of CDKL2 in normal and tumor tissues were analyzed by methylation-sensitive restriction enzyme-coupled real-time quantitative PCR(MSRE-qPCR).Quantitative real-time PCR(qPCR)was used to detect the difference of CDKL2 expression in normal and tumor tissues,and to evaluate the correlation between CDKL2 methylation and mRNA expression levels.The effect of CDKL2 methylation on the proliferation and migration of glioblastoma cells was verified in vitro and in vivo and tested by the methylation inhibitor 5-aza-2′-deoxycytidine.Results:Receiver characteristic curve(ROC)analysis showed high specificity and sensitivity of CDKL2 methylation in non-tumor and glioma tissues(P<0.001).CDKL2 showed high methylation and low expression in glioma tissues compared with normal tissues(P<0.001),and the methylation level of CDKL2 was inversely proportional to the expression level.5-aza-2′-deoxycytidine effectively inhibited tumor formation and tumor cell growth(P<0.01).Conclusion:CDKL2 shows a high methylation level in glioma cells,and demethylation can inhibit the growth of tumor cells,which may be a potential target for glioma treatment.