HHV-6A感染对神经胶质瘤细胞LncRNA MEG3表达及增殖凋亡侵袭转化的影响  

Human herpesvirus 6A subtype affects the expression of LncRNA MEG3,proliferation,apoptosis and EMT process in glioma cells

作　　者：万昕 胡月[2] 章菊 陈云[3] 金佩佩[1] WAN Xin;HU Yue;ZHANG Ju;CHEN Yun;JIN Peipei(Department of Laboratory Medicine, The First Affiliated Hospital of University of Science and Technology of China/Anhui Provincial Hospital, Hefei 230036;Prenatal Diagnosis Center, The First Affiliated Hospital of USTC/Anhui Provincial Hospital, Hefei 230036, Anhui;Department of Microbiology and Immunology, Nanjing Medical University, Nanjing 211100, Jiangsu, China)

机构地区：[1]中国科学技术大学附属第一医院/安徽省立医院检验科,安徽合肥230036 [2]中国科学技术大学附属第一医院/安徽省立医院妇产科产前诊断中心,安徽合肥230036 [3]南京医科大学微生物与免疫学系,江苏南京211100

出　　处：《癌变．畸变．突变》2019年第5期352-358,共7页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金青年科学基金项目(81701823)

摘　　要：目的:选取胶质瘤相关长链非编码RNA(LncRNA)MEG3,观察人类疱疹病毒6A亚型(HHV-6A)感染对神经胶质瘤细胞LncRNA MEG3表达及其增殖、凋亡、侵袭、转化的影响.方法:实时定量PCR检测人神经胶质瘤细胞和正常神经胶质细胞,以及37例胶质瘤组织、4例癌旁组织、18例正常脑组织中LncRNA MEG3的表达.细胞计数试剂盒检测HHV-6A感染后神经胶质瘤细胞吸光度D(450)值,流式细胞术检测细胞凋亡率,划痕实验观察细胞划痕愈合度,蛋白质印迹法检测细胞上皮间质转化(EMT)指标蛋白的表达水平.结果:在胶质瘤组织或细胞中,LncRNA MEG3的表达水平较正常对照组织或细胞低(P<0.05),而HHV-6A感染后的正常神经胶质细胞或胶质瘤细胞中LncRNA MEG3的表达也呈降低趋势(P<0.05).流式细胞术结果显示病毒感染后胶质瘤细胞的凋亡率为(6.05±0.40)%,对照组细胞凋亡率为(8.23±0.75)%,HHV-6A感染组凋亡率较对照组显著降低(P<0.05).划痕实验结果显示HHV-6A感染后细胞划痕愈合度[(72.33±6.90)%]大于对照组细胞划痕愈合度[(43.67±6.30)%],差异有统计学意义(P<0.05).Western blot结果显示,与对照组相比,病毒感染后细胞中Snail及Vimentin蛋白表达水平升高,E-cadherin蛋白的表达降低(P<0.05).结论:HHV-6A感染可下调神经胶质细胞及胶质瘤细胞中LncRNA MEG3的表达,促进胶质瘤细胞增殖侵袭和上皮间质转化,并抑制其凋亡.OBJECTIVE:The glioma-associated long non-coding RNA (LncRNA) MEG3 was preliminarily selected and detected in glioma tissues. Its expression in glioma cells together with proliferation, apoptosis, invasion and transformation of glioma cells were investigated after HHV-6A infection. METHODS:Expression of LncRNA MEG3 in 37 glioma, 4 para-tumor tissues and 18 normal brain tissues, or HHV-6A infected glioma cells were measured by Real-time quantitative PCR. The D(450) value of HHV-6A infected glioma cells were assayed by using cell counting kit-8 method. The frequencies of apoptotic cells were detected by flow cytometry. The cell healing ability was monitored by the wound-healing assay. The epithelial-mesenchymal transition (EMT)-related proteins were evaluated by Western blot. RESULTS: Expression of LncRNA MEG3 was significantly lower in glioma tissues than in para-tumor tissues (P<0.05). Expression of LncRNA MEG3 showed a down-regulation in glioma or normal glial cells after HHV-6A infection (P<0.05). The apoptotic rates of HHV-6A infected glioma cells were significantly lower than that in the control group [(6.05±0.40)% and (8.23±0.75)%, respectively, P<0.05]. The wound healing ability of glioma cells after HHV-6A infection was significantly better than that of the control group [(72.33±6.90)% and (43.67±6.30)%, respectively, P<0.05].Compared with the control group, expression of Snail and Vimentin were increased, while expression of Ecadherin was decreased in glioma cells after HHV-6A infection (P<0.05). CONCLUSION:HHV-6A infection down-regulated the expression of LncRNA MEG3 in glial cells and glioma cells. The expression was associated with proliferation,invasion,epithelial-mesenchymal transition of glioma cells and inhibition of apoptosis.

关 键 词：神经胶质瘤 长链非编码RNAMEG3 人类疱疹病毒6A亚型 上皮间质转化 

分 类 号：R730.23[医药卫生—肿瘤]