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

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。

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

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.