中文摘要：本文对大鼠全部或选定组织中特异性表达谱进行表征以探讨转基因玉米MON810的安全性。作者利用新一代测序（NGS）和RT-qPCR技术对90天饲喂转基因玉米的大鼠肠组织的基因表现进行评价。喂养大鼠的饲料中分别含有33%的转基因玉米MON810和近等基因玉米。本文利用RT-qPCR方法研究阵列靶向凋亡、DNA损伤修复、未折叠蛋白反应（UPR）；利用NGS进行大鼠全部回肠组织分析。研究发现，在雄性和雌性大鼠组织对转基因饮食无生物学反应。RNA-seq基因表达的全转录组分析证实了这些发现。通过肠道组织的转录组分析来预测转基因食品/饲料潜在的意外影响，是一种补充经典毒理学试验研究的新方法。检测毒性试验过程中信号通路的改变可能增强毒理学试验结果的可信度。另外，在饲喂转基因玉米MON810的大鼠肠道表达谱中没有与转基因相关的显著变化，且指向饮食肠毒性的与细胞通路相关的变化（细胞凋亡、DNA损伤修复，UPR）均未观察到。
外文摘要：Background: Global as well as specific expression profiles of selected rat tissues were characterized to assess the safety of genetically modified (GM) maize MON810 containing the insecticidal protein Cry1Ab. Gene expression was evaluated by use of Next Generation Sequencing (NGS) as well as RT-qPCR within rat intestinal tissues based on mandatory 90-day rodent feeding studies. In parallel to two 90-day feeding studies, the transcriptional response of rat tissues was assessed as another endpoint to enhance the mechanistic interpretation of GM feeding studies and/or to facilitate the generation of a targeted hypothesis. Rats received diets containing 33% GM maize (MON810) or near-isogenic control maize. As a site of massive exposure to ingested feed the transcriptomic response of ileal and colonic tissue was profiled via RT-qPCR arrays targeting apoptosis, DNA-damage/repair, unfolded protein response (UPR). For global RNA profiling of rat ileal tissue, we applied NGS. Results: No biological response to the GM-diet was observed in male and in female rat tissues. Transcriptome wide analysis of gene expression by RNA-seq confirmed these findings. Nevertheless, gene ontology (GO) analysis clearly associated a set of distinctly regulated transcripts with circadian rhythms. We confirmed differential expression of circadian clock genes using RT-qPCR and immunoassays for selected factors, thereby indicating physiological effects caused by the time point of sampling. Conclusion: Prediction of potential unintended effects of GM-food/feed by transcriptome based profiling of intestinal tissue presents a novel approach to complement classical toxicological testing procedures. Including the detection of alterations in signaling pathways in toxicity testing procedures may enhance the confidence in outcomes of toxicological trials. In this study, no significant GM-related changes in intestinal expression profiles were found in rats fed GM-maize MON810. Relevant alterations of selected cellular pathways (apoptosis, DNA damage and repair, UPR) pointing toward intestinal toxicity of the diets were not observed. Transcriptomic profiles did not reveal perturbations of pathways associated with toxicity, underlining the study results revealed by classical OECD endpoints.
外文关键词：transcriptomics profiles; pathway-analysis; rat feeding-trial; GM-plant; MON810-maize
作者：Sharbati, Jutta; Bohmer, Marc; Bohmer, Nils; 等.
作者单位：Free Univ Berlin
期刊名称：FRONTIERS IN GENETICS
期刊影响因子：3.789
出版年份：2017
出版刊次：12
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