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Whole transcriptome sequencing of gastric cancer stem cells and non-gastric cancer stem cells

ABSTRACT: As a subpopulation of tumor cells with stemness characteristics such as self-renewal ability and high oncogenicity, gastric cancer stem cells (GCSCs) are considered to be tumor-initiating cells and are the root cause of cancer recurrence and chemoresistance. Therefore, an in-depth study of the stemness characteristics and maintenance mechanism of GCSCs is of great significance for the treatment of gastric cancer (GC). In this study, GCSCs and non-gastric cancer stem cells (Non-GCSCs) derived from tumor tissues of GC patients were used for whole transcriptome sequencing. The goal of identifying key differentially expressed circRNAs, lncRNAs, miRNAs and mRNAs associated with GCSCs. In conclusion, we mapped the molecular feathers at RNA level closely related to GCSCs and revealed certain ncRNAs with great research potentia based on the whole transcriptome sequencing.

ORGANISM(S): Homo sapiens

PROVIDER: GSE274738 | GEO | 2025/11/14

REPOSITORIES: GEO

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Project description:Background: Helicobacter pylori (HP) infection may initiate and promote progression of gastric carcinogenesis. ONECUT2 shows promise for tumor diagnosis, prognosis, and treatment. This study explored ONECUT2's role and specific mechanism underlying HP infection-associated gastric carcinogenesis to suggest a basis for targeting ONECUT2 as a therapeutic strategy for gastric cancer (GC). Methods: Public data, single-cell RNA sequencing, spatial transcriptome analysis, RNA sequencing, GC tissue specimens, and clinical survival data were analyzed. Human GC organoids, HP infection, 3D sphere-forming, and extreme-dilution nude mouse models were constructed. Western blotting, qPCR, immunohistochemical staining, dual-luciferase reporter assays, phosphokinase microarray, and immunofluorescence were used. Results: Multidimensional data supported an association between ONECUT2, HP infection, and GC pathogenesis. HP infection upregulated ONECUT2 transcriptional activity via NFκB. In vitro and in vivo experiments demonstrated that ONECUT2 increases stemness in GC cells. ONECUT2 was also shown to inhibit PPP2R4 transcription, resulting in reduced PP2A activity, which in turn increased AKT/β-catenin phosphorylation. AKT/β-catenin phosphorylation facilitates β-catenin translocation to the nucleus, initiating transcription of downstream stemness-associated genes in GC cell. HP infection could upregulate the phosphorylation of AKT and β-catenin triggered by ONECUT2 downregulation via induction of ONECUT2. Clinical survival analysis indicated that high ONECUT2 expression might be an indicator of poor prognosis in GC. Conclusion: This study highlights a critical role played by ONECUT2 in promoting HP infection-associated GC by enhancing cell stemness through the PPP2R4/AKT/β-catenin signaling pathway. These findings suggest promising therapeutic strategies and potential therapeutic targets for GC treatment. Keywords: Helicobacter pylori; gastric cancer; prognosis; tumor stemness; ONECUT2

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Whole transcriptome sequencing of gastric cancer stem cells and non-gastric cancer stem cells

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