摘要
[目的]探讨良性脑膜瘤中炎症因子IL-1β在NF2甲基化中的作用及调控机制。[方法]利用2个脑膜瘤患者手术后的标本做原代细胞培养,不同浓度IL-1β(0,0.2,0.4,0.8,1.0,10ng/ml)处理细胞,利用RT-PCR和Western blot检测NF2、merlin和DNMTs的表达,利用甲基化特异性PCR检测NF2启动子甲基化水平。通过共同添加IL-1β和DNMT抑制剂或siRNA,观察IL-1β与NF2甲基化的关系。[结果] IL-1β能促进脑膜瘤细胞的增殖。RT-PCR及Western blot结果显示,IL-1β能降低脑膜瘤细胞NF2和merlin的表达。IL-1β能诱导脑膜瘤细胞NF2启动子甲基化;同时IL-1β能增加脑膜瘤细胞中DNMT1的表达,而IL-1β诱导脑膜瘤细胞的NF2启动子甲基化可以被DNMT抑制剂和siRNA抑制。[结论 ]IL-1β通过DNMT1诱导NF2基因启动子的甲基化,这种甲基化可以下调NF2基因的转录。抑制DNMTs可以考虑作为调控NF2表达和和抑制脑膜瘤发展的治疗靶点。
[Objective] To investigate the role of IL-1β in regulating the NF2 gene methylation in benign meningioma. [Methods] The primary meningioma cells obtained from 2 patients were cultured,and the cells were treated with IL-1β at different concentrations(0,0.2,0.4,0.8,1.0,10ng/ml).The m RNA and protein expressions of NF2,merlin and DNMT were detected by RT-PCR and Western blot,respectively. The methylation level of NF2 promoter was detected by MS-PCR. The relationship between IL-1β and NF2 methylation was observed by co-treatment with IL-1β and DNMT inhibitors or siRNA. [Results] IL-1β promoted the proliferation of meningioma cells. RTPCR and Western blot results showed that IL-1β reduced the expression of NF2 and Merlin in meningioma cells. IL-1β induced methylation of NF2 gene promoter in meningioma cells,and increased the expression of DNMT1 in meningioma cells,while this effect was inhibited by DNMT inhibitors and siRNA. [Conclusion] IL-1β induces methylation of NF2 gene promoter through DNMT1,which can downregulate the transcription of NF2 gene. Inhibition of DNMTs might be used as a therapeutic approach for regulating the expression of NF2 and inhibiting the development of meningioma.
作者
李峰
张超
LI Feng;ZHANG Chao(No.215 Hospital of Shaanxi Nuclear Industry,Xianyang 712000,China;Hanzhong Central Hospital,Hanzhong 723000,China)
出处
《肿瘤学杂志》
CAS
2019年第2期116-120,共5页
Journal of Chinese Oncology
