Institute of Cytology of the Russian Academy of Science, 194064 St-Petersburg, Russia.
Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, 1015 Walnut Street, Ste. 1024, Philadelphia, PA 19107, USA.
Int J Mol Sci. 2021 Apr 28;22(9):4682. doi: 10.3390/ijms22094682.
Cancer stem cells' (CSCs) self-maintenance is regulated via the pluripotency pathways promoting the most aggressive tumor phenotype. This study aimed to use the activity of these pathways for the CSCs' subpopulation enrichment and separating cells characterized by the OCT4 and SOX2 expression.
To select and analyze CSCs, we used the SORE6x lentiviral reporter plasmid for viral transduction of colon adenocarcinoma cells. Additionally, we assessed cell chemoresistance, clonogenic, invasive and migratory activity and the data of mRNA-seq and intrinsic disorder predisposition protein analysis (IDPPA).
We obtained the line of CSC-like cells selected on the basis of the expression of the OCT4 and SOX2 stem cell factors. The enriched CSC-like subpopulation had increased chemoresistance as well as clonogenic and migration activities. The bioinformatic analysis of mRNA seq data identified the up-regulation of pluripotency, development, drug resistance and phototransduction pathways, and the downregulation of pathways related to proliferation, cell cycle, aging, and differentiation. IDPPA indicated that CSC-like cells are predisposed to increased intrinsic protein disorder.
The use of the SORE6x reporter construct for CSCs enrichment allows us to obtain CSC-like population that can be used as a model to search for the new prognostic factors and potential therapeutic targets for colon cancer treatment.
癌症干细胞（CSCs）的自我维持是通过促进最具侵袭性肿瘤表型的多能性途径来调节的。本研究旨在利用这些途径的活性来富集 CSCs 亚群，并分离出具有 OCT4 和 SOX2 表达特征的细胞。
为了选择和分析 CSCs，我们使用了 SORE6x 慢病毒报告质粒对结肠腺癌细胞进行病毒转导。此外，我们评估了细胞的化学抗性、克隆形成、侵袭和迁移活性以及 mRNA-seq 和固有无序倾向蛋白分析（IDPPA）的数据。
我们获得了基于 OCT4 和 SOX2 干细胞因子表达选择的 CSC 样细胞系。富集的 CSC 样亚群具有更高的化学抗性以及克隆形成和迁移活性。mRNA-seq 数据的生物信息学分析表明，多能性、发育、耐药性和光转导途径上调，而与增殖、细胞周期、衰老和分化相关的途径下调。IDPPA 表明 CSC 样细胞易发生内在蛋白无序增加。
使用 SORE6x 报告构建体富集 CSCs 可使我们获得 CSC 样群体，可将其用作搜索结肠癌治疗新的预后因素和潜在治疗靶点的模型。
