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在 3D 胶原中进行高通量转录组和蛋白质组学分析以研究间充质-阿米巴样转化。

High-throughput transcriptomic and proteomic profiling of mesenchymal-amoeboid transition in 3D collagen.

机构信息

Department of Cell Biology, Charles University, Viničná 7, Prague, Czech Republic.

Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic.

出版信息

Sci Data. 2020 May 27;7(1):160. doi: 10.1038/s41597-020-0499-2.

Abstract

The plasticity of cancer cell invasion represents substantial hindrance for effective anti-metastatic therapy. To better understand the cancer cells' plasticity, we performed complex transcriptomic and proteomic profiling of HT1080 fibrosarcoma cells undergoing mesenchymal-amoeboid transition (MAT). As amoeboid migratory phenotype can fully manifest only in 3D conditions, all experiments were performed with 3D collagen-based cultures. Two previously described approaches to induce MAT were used: doxycycline-inducible constitutively active RhoA expression and dasatinib treatment. RNA sequencing was performed with ribo-depleted total RNA. Protein samples were analysed with tandem mass tag (TMT)-based mass spectrometry. The data provide unprecedented insight into transcriptome and proteome changes accompanying MAT in true 3D conditions.

摘要

肿瘤细胞侵袭的可塑性是有效抗转移治疗的主要障碍。为了更好地了解肿瘤细胞的可塑性,我们对 HT1080 纤维肉瘤细胞进行了复杂的转录组和蛋白质组分析,这些细胞经历了间质-阿米巴样过渡(MAT)。由于阿米巴样迁移表型只能在 3D 条件下完全表现出来,因此所有实验都是在基于 3D 胶原的培养物中进行的。使用了两种先前描述的方法来诱导 MAT:诱导型组成性激活 RhoA 表达和 dasatinib 处理。用核糖体耗尽的总 RNA 进行 RNA 测序。用串联质量标签(TMT)-基于质谱法分析蛋白质样品。这些数据提供了前所未有的见解,揭示了真正 3D 条件下 MAT 伴随的转录组和蛋白质组变化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0a6e/7253430/1216de8bbea8/41597_2020_499_Fig1_HTML.jpg

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