人疱疹病毒6型感染神经胶质瘤细胞中miR-301a表达的变化及生物学意义  

Expression and Biological Significance of miR-301a in Glioma Cells Infected with Human Herpesvirus 6

作　　者：王锋存[1] 刘军莉[1] 双杰[2] WANG Fengcun;LIU Junli;SHUANG Jie(Qinghai University Affiliated Hospital,Xining,810001,China;Teaching and Research Section Department of Pathogenic Biology,Medical College of Qinghai University,Xining,810001,China)

机构地区：[1]青海大学附属医院,西宁810001 [2]青海大学医学院病原生物学教研室,西宁810001

出　　处：《病毒学报》2022年第1期129-138,共10页Chinese Journal of Virology

摘　　要：人疱疹病毒6A型(Human Herpesvirus 6A,HHV6A)感染与神经胶质瘤的发病有关,但具体机制尚不清楚。HHV6A感染能够增加miR-301a的表达,而miR-301a表达在胶质瘤发病中起促进作用。为了观察HHV6A感染神经胶质瘤细胞中miR-301a表达的变化及生物学意义,本研究培养了神经胶质瘤细胞株U87、U251、U373,HHV6A感染后检测了miR-301a及RUNX3(Runt-Related Transcription Factor 3,RUNX3)的表达;U87细胞在感染HHV6A的同时分别转染阴性对照(Negative Control,NC)miR、miR301a抑制物、NC siRNA、RUNX3 siRNA,检测细胞增殖、迁移、侵袭;采用双荧光素酶报告基因实验验证miR-301a靶向RUNX3。结果显示,HHV6感染的U87、U251、U373细胞中miR-301a表达增加、RUNX3表达降低且HHV6感染的U87中miR-301a上调、RUNX3下调幅度最大。在U87细胞中,与对照组比较,HHV6组增殖、迁移、侵袭增强,RUNX3表达降低;与miR-NC+HHV6组比较,miR-301a敲低+HHV6组增殖、迁移、侵袭减弱,RUNX3表达增加;与HHV6+miR-301a敲低+si-NC组比较,HHV6+miR-301a敲低+si-RUNX3组增殖、迁移、侵袭增强,RUNX3表达降低;经双荧光素酶报告基因验证,miR-301a靶向RUNX3基因mRNA 3’UTR。结果表明,HHV6A感染神经胶质瘤细胞中miR-301a表达增加,进而通过靶向抑制RUNX3促进细胞增殖、迁移、侵袭。本研究初步探究了HHV6A感染促进神经胶质瘤细胞增殖、迁移、侵袭的作用及分子机制,进而为深入认识神经胶质瘤恶性生物学行为的调控机制及发病机制提供了实验依据,也为今后发现神经胶质瘤新的防治靶点提供思路。Human herpesvirus 6 A(HHV6 A)infection is associated with glioma pathogenesis,but the specific mechanism is not known.HHV6 A infection can increase microRNA(mir)-301 a expression,which plays an important part in glioma pathogenesis.To measure the expression and biological significance of miR-301 a in glioma cells infected with HHV6 A,three glioma cell lines(U87,U251,and U373)were cultured.Expression of miR-301 a and Runt-related transcription factor(RUNX)3 after HHV6 A infection was measured.Simultaneously with infection by HHV6 A,U87 cells were transfected with negative control(NC)miR,a miR-301 a inhibitor,NC small interfering(si)RNA,and RUNX3 siRNA.Then,the proliferation,migration,and invasion of glioma cells were detected.A double-luciferase reporter gene assay was used to verify that miR-301 a was targeting RUNX3.We discovered that miR-301 a expression was increased and RUNX3 expression was decreased in U87,U251 and U373 cells infected with HHV6.Upregulation of miR-301 a expression and downregulation of RUNX3 expression in U87 cells infected with HHV6 were the most significant.Compared with the control group,the proliferation,migration,and invasion of glioma cells were enhanced,and RUNX3 expression decreased,in the HHV6 group.Compared with the miR-NC+HHV6 group,the proliferation,migration,and invasion of glioma cells were weakened,and RUNX3 expression increased in the miR-301 a knockdown+HHV6 group.Compared with the HHV6+miR-301 a knockdown+si-NC group,the proliferation,migration,and invasion of glioma cells were enhanced,and RUNX3 expression decreased in the HHV6+miR-301 a knockdown+si-RUNX3 group.The double-luciferase reporter gene showed that miR-301 a targeted the mRNA 3’UTR of RUNX3.These results indicate that miR-301 a expression increases in glioma cells infected with HHV6 A,which promotes the proliferation,migration,and invasion of cells by targeting RUNX3.In a preliminarily fashion,we explored the role and molecular mechanism of HHV6 A infection in promoting the proliferation,migration,and invasion

关 键 词：神经胶质瘤 人疱疹病毒6A型(HHV6A) miR-301a RUNX3 增殖 迁移 侵袭 

分 类 号：R373.9[医药卫生—病原生物学]