神经胶质瘤细胞 ATF5表达水平对 HCMV 复制的影响

摘要

The replication levels of human cytomegalovirus(HCMV)in gliomata are different,and the mechanism is still unknown. In this study the changes of replication levels of the virus after its infection of gliomata were detected through down-regulation of the expression of activation transcription factor 5(ATF5)in gliomata. Firstly,cells U87, SY5Y,and A172 were infected with HCMV respectively,and observe the changes of their cellular morphology,and collect each of their supernatants at the time points of 24,48,72,96,and 120 h to test their viral titers,and detect the expression situation of incontinent incipient gene IE2,incipient gene UL44,terminal gene UL99,and ATF5 with real-time PCR,and detect the expression situation of the HCMV coded protein and ATF5 with Western-blot. The re-sults showed that as compared the replication level of HCMV in cells A172 with the ones in cells U87 and SY5Y,the group of U87 and SY5Y cells were significantly higher than that of the replication level in the group of A172 cells (P < 0. 05);and as compared the expression of ATF5 in the groups of U87 cell and SY5Y cell were significantly high-er than the group of A172(P < 0. 05);Using lentivirus mediation RNA interfering technique to down-regulate the ex-pression of ATF5 in U87 and SY5Y,using HCMV infected cells to detect the expression of viral gene and protein,the results showed that the down-regulation of the expression of ATF5 could inhibit the replication of HCMV(P < 0. 05). The above results showed that down-regulate the expression level of ATF5 in glioma cell could inhibit the replication level of HCMV.%人巨细胞病毒（human cytomegalovirus，HCMV）在神经胶质瘤细胞中的复制水平不一，其机制尚不清楚。本研究通过下调转录激活因子5（ATF5）在神经胶质瘤细胞中的表达，检测 HCMV 感染神经胶质瘤细胞后病毒复制水平的变化。首先用 HCMV AD169（MOI =5）分别感染 U87、SY5Y 及 A172细胞，观察细胞形态变化，分别在24、48、72、96、120 h 取各时间点上清液检测病毒滴度；Real-time PCR 检测 HCMV 即刻早期基因（IE2）、早期基因（UL44）、晚期基因（UL99）及 ATF5的表达情况；Western-blot 检测病毒基因编码蛋白及 ATF5表达的情况。结果显示 HCMV 在 U87、SY5Y 细胞中复制水平与病毒在 A172细胞中复制水平相比，U87、SY5Y细胞组明显高于 A172细胞组（P 约0.05），ATF5表达在 U87、SY5Y 细胞组与 A172细胞组相比，U87、SY5Y 细胞组 ATF5表达明显高于 A172组（P 约0.05）；利用慢病毒介导的 RNA 干扰技术下调 ATF5在 U87、SY5Y 细胞的表达，用 HCMV 感染细胞检测病毒基因及蛋白的表达，结果 ATF5表达下调可抑制 HCMV 的复制（P 约0.05）。以上结果表明，在胶质瘤细胞中下调 ATF5表达水平可以抑制 HCMV 的复制水平。