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暴露于 2,3,7,8-四氯二苯并对二恶英的猪颗粒细胞的转录谱分析。

Transcriptional profiling of porcine granulosa cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

机构信息

Department of Animal Physiology, Faculty of Biology and Biotechnology, University of Warmia and Mazury in Olsztyn, Poland.

Laboratory of Molecular Diagnostics, Faculty of Biology and Biotechnology, University of Warmia and Mazury in Olsztyn, Poland.

出版信息

Chemosphere. 2017 Jul;178:368-377. doi: 10.1016/j.chemosphere.2017.03.055. Epub 2017 Mar 16.

Abstract

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a toxic man-made chemical compound contaminating the environment. An exposure of living organisms to TCDD may result in numerous disorders, including reproductive pathologies. The aim of the current study was to examine the effects of TCDD on the transcriptome of porcine granulosa cell line AVG-16. By employing next-generation sequencing (NGS) we aimed to identify genes potentially involved in the mechanism of TCDD action and toxicity in porcine granulosa cells. The AVG-16 cells were treated with TCDD (100 nM) for 3, 12 or 24 h, and afterwards total cellular RNA was isolated and sequenced. In TCDD-treated cells we identified 141 differentially expressed genes (DEGs; p < 0.05 and log2 fold change ≥1.0). The DEGs were assigned to GO term, covering biological processes, molecular functions and cellular components. Due to the large number of genes with altered expression, in the current study we analyzed only the genes involved in follicular growth, development and functioning. The obtained results showed that TCDD may affect ovarian follicle fate by influencing granulosa cell cycle, proliferation and DNA repair. The demonstrated over-time changes in the quantity and quality of genes being affected by TCDD treatment showed that the effects of TCDD on granulosa cells changed dramatically between 3-, 12- and 24-h of cell culture. This finding indicate that timing of gene expression measurement is critical for drawing correct conclusions on detailed relationships between the TCDD-affected genes and resulting intracellular processes. These conclusions have to be confirmed and extended by research involving proteomic and functional studies.

摘要

2,3,7,8-四氯二苯并对二恶英(TCDD)是一种有毒的人造化学化合物,污染环境。生物体暴露于 TCDD 中可能导致多种疾病,包括生殖系统病理学。本研究的目的是研究 TCDD 对猪颗粒细胞系 AVG-16 转录组的影响。通过采用下一代测序(NGS),我们旨在鉴定可能参与 TCDD 作用机制和猪颗粒细胞毒性的基因。将 AVG-16 细胞用 TCDD(100 nM)处理 3、12 或 24 小时,然后分离并测序总细胞 RNA。在 TCDD 处理的细胞中,我们鉴定了 141 个差异表达基因(DEGs;p < 0.05,log2 倍数变化≥1.0)。这些 DEGs 被分配到 GO 术语,涵盖了生物过程、分子功能和细胞成分。由于表达改变的基因数量众多,在当前研究中,我们仅分析了参与卵泡生长、发育和功能的基因。研究结果表明,TCDD 可能通过影响颗粒细胞周期、增殖和 DNA 修复来影响卵巢卵泡命运。TCDD 处理后基因数量和质量的变化表明,TCDD 对颗粒细胞的影响在细胞培养 3、12 和 24 小时之间发生了巨大变化。这一发现表明,在得出关于 TCDD 影响的基因与细胞内过程之间的详细关系的正确结论时,测量基因表达的时间非常关键。这些结论必须通过涉及蛋白质组学和功能研究的研究来证实和扩展。

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