NF2基因在丙肝病毒非结构蛋白4B调节细胞周期中的作用

Role of NF2 gene in the regulation of cell cycle mediated by HCV NS4B

目的在HEPG2细胞内观察丙肝病毒(HCV)非结构蛋白4B(NS4B)对神经纤维瘤病Ⅱ型(NF2)基因的影响及NF2在NS4B调节细胞周期中的作用。方法将细胞分为多组,其中3组对照,分别为无处理组、pCDNA3.1空质粒组和si-NF2 control组;3组作用组,分别为NS4B质粒组、NF2质粒组和si-NF2沉默基因组。每组经过转染、培养后收获细胞;RT-qPCR和Western blot检测细胞内相应基因转录及表达;流式细胞仪检测细胞周期。结果NS4B可降低NF2转录水平(P<0.01),同时NS4B的高表达和NF2的低表达对细胞周期具有相似的作用,及促进细胞周期向S期的转变;而NF2的高表达则使细胞周期停滞在G 0/G 1。结论NS4B对NF2具有一定的调控作用,NF2可能参与NS4B调控细胞周期机制。

Objective To investigate the effect of HCV nonstructural protein 4 B(NS4 B)on NF2 in HEPG2 cells and the role of NF2 in NS4 B regulation of cell cycle.Methods Cells were divided into blank control pCDNA3.1 plasmid group,NS4 B plasmid group,NF2 plasmid group and si-NF2 silencing gene group and si-control gene were transfected into other groups by cell transfection separately.RT-qPCR and Western blot were used to detect the transcription and expression of the corresponding genes.Flow cytometry was used to detect the cell cycle.ResultsThe expression of NF2 was decreased by NS4 B.The high expression of NS4 B and the low expression of NF2 promoted S phase change in cell cycle,while high expression of NF2 arrested cell cycle in G0/G1.ConclusionsNS4 B can regulate NF2 and NF2 might be involved in the mechanism of regulating cell cycle by NS4 B.