Isolation and Characterization of Human Colon Adenocarcinoma Stem-Like Cells Based on the Endogenous Expression of the Stem Markers

Abstract

Background: 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. Methods: 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). Results: 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. Conclusion: 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.

Polyploidy and Myc Proto-Oncogenes Promote Stress Adaptation via Epigenetic Plasticity and Gene Regulatory Network Rewiring

“…Under normoxia, polyploidy is usually accompanied by enhanced glycolysis, glutaminolysis, hypertranscription, active protein synthesis, and ribosome biogenesis, which are also manifestations of embryonality [ 2 , 6 , 71 , 79 , 186 , 187 ]. All these features of metabostemness have also been described in endopolyploid cells of Drosophila in development and wound healing, in cancer polyploid cells, and in cells of normal mammalian tissues [ 7 , 57 , 178 , 184 , 188 ].…”

“…Metabolic pathways of polyploid cells also demonstrate manifestations of stemness [ 11 , 71 , 72 ]. Similarly to pluripotent stem cells, polyploid cells can simultaneously derive energy from pathways that are incompatible with differentiated cells, promoting adaptation to stress associated with energy deprivation [ 49 , 72 , 176 , 177 , 178 , 179 , 180 , 181 , 182 ]. Recent data obtained from polyploid cells from normal tissues of animals and plants, various tumors, and yeasts indicate that these cells possess energetically flexible metabolism combining glycolysis glutaminolysis and oxidative phosphorylation [ 43 , 57 , 72 , 183 , 184 , 185 ].…”

Cancer Stem Cells and Their Vesicles, Together with Other Stem and Non-Stem Cells, Govern Critical Cancer Processes: Perspectives for Medical Development

“…Most often, however, they can be envisaged according to general criteria. CSCs share some properties with MSCs, including broad proliferation and activation of various signaling processes [ 18 , 19 , 20 , 34 ]. Relevant to CSCs are the cellular cross-talks also involving, together with stem cells, non-cancer cells accumulated at the niches [ 15 , 16 , 24 , 25 , 35 ] ( Table 1 ; Figure 1 ).…”

“…However, most key properties of CSCs are specific [ 17 , 34 ]; their modulation appears largely focused on self-renewal and multi-lineage differentiations [ 20 , 21 ], which is different from those of the MSC family. CSCs show multi-lineage properties, leading to the generation of distinct cancer subtypes [ 18 , 21 ].…”

“…In contrast, the non-small cell lung cancer appears reinforced [ 17 ]. Interestingly, the positive results induced by MSC in some tumors were not confirmed in their metastases where MSC responses were negative [ 15 , 34 ]. In this case, therefore, different mechanisms taking place in different areas of cancer pathology, appear to govern an apparently single process.…”