Department of Medical Biology, University of Pécs Medical School, Pécs, Hungary; Signal Transduction Research Group, János Szentágothai Research Centre, Pécs H-7624, Hungary.
Experimental and Clinical Research Center, Charité Medical Faculty and Max-Delbrück Center for Molecular Medicine, Berlin 13125, Germany.
Virus Res. 2014 Jun 24;185:10-22. doi: 10.1016/j.virusres.2014.03.003. Epub 2014 Mar 15.
Although the oncolytic potential of natural, non-engineered Newcastle disease virus (NDV) isolates are well-known, cellular mechanisms determining NDV sensitivity of tumor cells are poorly understood. The aim of the present study was to look for gene expression changes in PC12 pheochromocytoma cells infected with an attenuated NDV strain that may be related to NDV susceptibility. PC12 cells were infected with the NDV strain MTH-68/H for 12h at a titer corresponding to the IC₅₀ value. Total cytoplasmic RNA samples isolated from control and MTH-68/H-infected cells were analyzed using a rat specific Affymetrix exon chip. Genes with at least 2-fold increase or decrease in their expression were identified. MTH-68/H-induced gene expression changes of 9 genes were validated using quantitative reverse transcriptase PCR. A total of 729 genes were up- and 612 genes were down-regulated in PC12 cells infected with MTH-68/H. Using the DAVID functional annotation clustering tool, the up- and down-regulated genes can be categorized into 176 and 146 overlapping functional gene clusters, respectively. Gene expression changes affecting the most important signaling mechanisms (Toll-like receptor signaling, RIG-I-like receptor signaling, interferon signaling, interferon effector pathways, apoptosis pathways, endoplasmic reticulum stress pathways, cell cycle regulation) are analyzed and discussed in detail in this paper. NDV-induced gene expression changes described in this paper affect several regulatory mechanisms and dozens of putative key proteins that may determine the NDV susceptibility of various tumors. Further characterization of these proteins may identify susceptibility markers to predict the chances of virotherapeutic treatment of human tumors.
虽然天然、非工程化的新城疫病毒（NDV）分离株的溶瘤潜力是众所周知的，但决定肿瘤细胞对 NDV 敏感性的细胞机制还知之甚少。本研究旨在寻找感染弱毒 NDV 株的 PC12 嗜铬细胞瘤细胞中可能与 NDV 易感性相关的基因表达变化。PC12 细胞以相当于 IC₅₀ 值的滴度感染 MTH-68/H NDV 株 12 小时。从对照和 MTH-68/H 感染的细胞中分离的总细胞质 RNA 样品使用大鼠特异性 Affymetrix 外显子芯片进行分析。鉴定出表达至少增加或减少 2 倍的基因。使用定量逆转录 PCR 验证了 MTH-68/H 诱导的 9 个基因的表达变化。MTH-68/H 感染的 PC12 细胞中，共有 729 个基因上调，612 个基因下调。使用 DAVID 功能注释聚类工具，上调和下调的基因可分别归类为 176 和 146 个重叠的功能基因簇。本文详细分析和讨论了影响最重要的信号转导机制（Toll 样受体信号转导、RIG-I 样受体信号转导、干扰素信号转导、干扰素效应途径、细胞凋亡途径、内质网应激途径、细胞周期调控）的基因表达变化。本文描述的 NDV 诱导的基因表达变化影响了几十个可能决定各种肿瘤对 NDV 敏感性的调节机制和假定的关键蛋白。对这些蛋白质的进一步表征可能会确定易感性标志物，以预测人类肿瘤病毒治疗的机会。
