人疱疹病毒6A型感染神经胶质瘤细胞中RUNX3与miR-301a表达的相关性及意义  

Correlation and Significance of Expression of RUNX3 and miR-301a in Glioma Cells Infected with Human Herpesvirus 6A

作　　者：王年臻 余丹[1] 杨国帅[1] 周治平[1] 侯丹[1] 胡裕洁 WANG Nianzhen;YU Dan;YANG Guoshuai;ZHOU Zhiping;HOU Dan;HU Yujie(Haikou Hospital affiliated to Xiangya Medical College,Central South University,Haikou 570208,China)

机构地区：[1]中南大学湘雅医学院附属海口医院,海口570208

出　　处：《病毒学报》2023年第3期775-782,共8页Chinese Journal of Virology

摘　　要：研究HHV-6A感染神经胶质瘤细胞中RUNX3与miR-301a表达的相关性及意义。神经胶质瘤细胞株U251、U87、U373感染HHV-6A,检测RUNX3、miR-301a的相对表达水平;U87细胞进行分组,感染HHV-6A,转染阴性对照(NC)miR模拟物、miR-301a模拟物、NC miR抑制物或miR-301a抑制物,检测细胞增殖A450水平、迁移数目、侵袭数目、miR-301a、RUNX3、c-myc、E-cadherin、MMP2、MMP9的相对表达水平、RUNX3野生型和突变型报告基因的荧光活力。研究结果显示HHV-6A感染后U251、U87及U373细胞中RUNX3的mRNA相对表达水平均降低、miR-301a相对表达水平均增加,其中U87细胞中RUNX3、miR-301a表达变化最显著。miR-301a组U87细胞中RUNX3野生型报告基因的荧光活力均低于miR-NC组(P<0.05),RUNX3突变型报告基因的荧光活力与miR-NC组比较无差异(P>0.05);HHV-6A感染的同时转染miR抑制物,miR-NC抑制物+HHV-6A组U87细胞的A450水平、迁移数目、侵袭数目、miR-301a、c-myc、MMP2、MMP9的相对表达水平均高于miR-NC抑制物组,RUNX3、Ecadherin的相对表达水平均低于miR-NC抑制物组(P<0.05);miR-301a抑制物+HHV-6A组U87细胞的A450水平、迁移数目、侵袭数目、miR-301a、c-myc、MMP2、MMP9的相对表达水平均低于miR-NC抑制物+HHV-6A组,RUNX3、E-cadherin的相对表达水平均高于miR-NC抑制物+HHV-6A组(P<0.05)。综上,HHV-6A感染促进神经胶质瘤细胞的增殖、迁移、侵袭,这一促进作用与增加miR-301a表达、抑制RUNX3表达有关。The regulatory effect and mechanism of action of human herpesvirus type 6A(HHV-6A)on glioma cells are not clear.Glioma cell lines(U251,U87,U373)were cultured.Relative expression of RUNX family transcription factor 3(RUNX3)and microRNA(miR)-30la after HHV-6A infection was measured.U87 cells with the most significant changes in expression of RUNX3 and miR-30la after HHV-6A infection were selected and grouped,infected with HHV-6A,transfected with the negative control(NC)miR mimic,miR-30la mimic,NC miR inhibitor or miR-30la inhibitor.We measured the proliferation,migration,the invasion of A450 cells.We also measured the relative expression of miR-30la,RUNX3,c-myc,E-cadherin,matrix metalloproteinase(MMP)2,MMP9,as well as the fluorescence intensity of RUNX3 wild-type and mutant reporter genes.After HHV-6A infection,the relative mRNA expression of RUNX3 decreased and relative expression of miR-30la increased in U251,U87 and U373 cells,among which expression of RUNX3 and miR-30la changed most significantly in U87 cells.The fluorescence intensity of the RUNX3 wild-type reporter gene in U87 cells in the miR-30la group was lower than that in the miR-NC group(P<0.05),and there was no significant difference in expression of the RUNX3 mutant reporter gene compared with that in the miR-NC group(P>0.05).We undertook infection with HHV-6A and transfection with an miR inhibitor simultaneously.We discovered that the proliferation,migration,and invasion of A450 cells,and relative expression of miR-30la,c-myc,MMP2 and MMP9 in U87 cells of the miR-NC inhibitor+HHV-6A group were higher than those of the miR-NC inhibitor group,and relative mRNA expression of RUNX3 and E-cadherin was lower than that of the miR-NC inhibitor group(P<0.05).Also,the proliferation,migration,and invasion of A450 cells,and relative expression of miR-30la,c-myc,MMP2 and MMP9 in U87 cells of the miR-30la inhibitor+HHV-6A group were lower than those of the miR-NC inhibitor+HHV-6A group,and relative expression of RUNX3 and E-cadherin was higher than that of miR-NC

关 键 词：神经胶质瘤 miR-301a RUNX3 增殖 迁移 侵袭 

分 类 号：R373.9[医药卫生—病原生物学]