BACKGROUND Pancreatic cancer is a fatal disease associated with resistance to conventional therapies. This study aimed to determine changes in gene expression patterns associated with infection and susceptibility of pancreatic cancer cells to an oncolyticvaccinia virus, GLV-1h153, carrying the human sodium iodide symporter for deep tissue imaging of virotherapy. METHODS Replication and susceptibility of pancreatic adenocarcinoma PANC-1 cells to GLV-1h153 was confirmed with replication and cytotoxicity assays. PANC-1 cells were then infected with GLV-1h153 and near-synchronous infection confirmed via flow cytometry of viral-induced green fluorescent protein (GFP) expression. Six and 24 hours after infection, three samples of each time point were harvested, and gene expression patterns assessed using HG-U133A cDNA microarray chips as compared to uninfected control. Differentially expressed genes were identified using Bioconductor LIMMA statistical analysis package. A fold change of 2.0 or above was used as a cutoff, with a P value of 0.01. The gene list was then analyzed using Ingenuity Pathways Analysis software. RESULTS Differential gene analysis revealed a total of 12,412 up- and 11,065 downregulated genes at 6 and 24 hours postinfection with GLV-1h153 as compared to control. At 6 hours postinfection. A total of 139 genes were either up or downregulated >twofold (false discovery rate < 0.05), of which 124 were mapped by Ingenuity Pathway Analysis (IPA). By 24 hours postinfection, a total of 5,698 genes were identified and 5,563 mapped by IPA. Microarray revealed gene expression changes, with gene networks demonstrating downregulation of processes such as cell death, cell cycle, and DNA repair, and upregulation of infection mechanisms (P < 0.01). Six hours after infection, gene changes involved pathways such as HMGB-1, interleukin (IL)-2, IL-6, IL-8, janus kinase/signal tranducer and activator of transcription (JAK/STAT), interferon, and ERK 5 signaling (P < 0.01). By 24 hours, prominent pathways included P53- and Myc-induced apoptotic processes, pancreatic adenocarcinoma signaling, and phosphoinositide 3-kinase/v-akt murine thymoma vial oncogene homolog 1 (PI3/AKT) pathways. CONCLUSIONS Our study reveals the ability to assess time-dependent changes in gene expression patterns in pancreatic cancer cells associated with infection and susceptibility to vaccinia viruses. This suggests that molecular assays may be useful to develop safer and more efficacious oncolyticvirotherapies and support the idea that these treatments may target pathways implicated in pancreatic cancer resistance to conventional therapies.

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与胰腺癌对溶瘤牛痘病毒疗法的敏感性相关的时间依赖性基因变化的分子网络，途径和功能分析。

背景技术胰腺癌是与常规疗法抗性相关的致命疾病。这项研究旨在确定与胰腺癌细胞对溶瘤牛痘病毒GLV-1h153的感染和易感性相关的基因表达模式的变化，该病毒携带人类碘化钠同向转运蛋白用于病毒疗​​法的深层组织成像。方法通过复制和细胞毒性试验确定胰腺腺癌PANC-1细胞对GLV-1h153的复制和敏感性。然后，用GLV-1h153感染PANC-1细胞，并通过流式细胞术确认病毒诱导的绿色荧光蛋白（GFP）表达，证实接近同步感染。感染后六个小时和24小时，在每个时间点采集了三个样本，与未感染对照相比，使用HG-U133A cDNA微阵列芯片评估基因表达模式。使用Bioconductor LIMMA统计分析工具包鉴定差异表达的基因。倍数变化为2.0或更高用作临界值，P值为0.01。然后使用Ingenuity Pathways Analysis软件分析基因清单。结果差异基因分析显示，与对照组相比，GLV-1h153感染后6和24小时共有12,412个上调基因和11,065个下调基因。感染后6小时。共有139个基因的表达上调或下调>两倍（错误发现率<0.05），其中124个通过Ingenuity Pathway Analysis（IPA）进行了定位。感染后24小时，通过IPA鉴定出总共5,698个基因并定位了5,563个基因。芯片揭示了基因表达的变化，基因网络显示出细胞死亡，细胞周期和DNA修复等过程的下调，以及感染机制的上调（P <0.01）。感染后六小时，基因变化涉及诸如HMGB-1，白介素（IL）-2，IL-6，IL-8，janus激酶/信号转导子和转录激活子（JAK / STAT），干扰素和ERK 5等途径。信号（P <0.01）。到24小时，突出的途径包括P53和Myc诱导的凋亡过程，胰腺腺癌信号传导和磷酸肌醇3-激酶/ v-akt鼠胸腺瘤小瓶癌基因同源物1（PI3 / AKT）途径。结论我们的研究揭示了评估与感染和牛痘病毒易感性相关的胰腺癌细胞基因表达模式随时间变化的能力。