View in EDS

Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells

Saved in:
Bibliographic Details
Title: Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells
Authors: Martins, Flávia, Machado, Ana L., Carvalho, Joana, Almeida, Catarina R., Beck, Hans C., Carvalho, Ana S., Backman, Vadim, Matthiesen, Rune, Velho, Sérgia
Source: Sci Rep
Scientific Reports, Vol 15, Iss 1, Pp 1-15 (2025)
Publisher Information: Springer Science and Business Media LLC, 2025.
Publication Year: 2025
Subject Terms: Proteomics, Science, Drug Resistance, Apoptosis, Article, Cell Line, Proto-Oncogene Proteins p21(ras)/genetics, Proto-Oncogene Proteins p21(ras), Apoptosis/genetics, Cell Line, Tumor, Humans, Gene Silencing, Wnt Signaling Pathway, Neoplastic, Tumor, Neoplasm/genetics, Unfolded Protein Response/genetics, HCT116 Cells, Colorectal Neoplasms/genetics, Gene Expression Regulation, Neoplastic, Gene Expression Regulation, Drug Resistance, Neoplasm, Unfolded Protein Response, Medicine, Colorectal Neoplasms
Description: Despite the development of mutant-selective KRAS inhibitors, colorectal cancer (CRC) responses remain limited, with stable disease and rapid recurrence being common outcomes. The molecular mechanisms enabling CRC cells to tolerate KRAS inhibition and ultimately develop resistance remain poorly understood. Here, we investigated early transcriptional and proteomic responses to KRAS silencing in 3D CRC cell line spheroid models, aiming to identify pathways associated with sensitivity or resistance to KRAS blockade. Cell lines were stratified into KRAS silencing-sensitive (HCT116 and SW480) and -resistant (LS174T and SW837) groups based on spheroid growth, cell cycle progression, and apoptosis induction. Transcriptional profiling revealed the unfolded protein response (UPR) and WNT/β-catenin signaling as pathways specifically upregulated in KRAS silencing-sensitive cells and downregulated in resistant cells. Proteomic analysis of membrane-enriched fractions further supported UPR deregulation, showing a pronounced downregulation of translation-related proteins in sensitive cells. Functional assays validated that the sensitive cell line HCT116 exhibits reduced protein aggregation and lower translational capacity upon KRAS knockdown, consistent with UPR activation. Pharmacological inhibition of IRE1α-mediated UPR signaling did not revert KRAS silencing-induced cell cycle arrest or apoptosis in this cell line. Collectively, our results highlight the UPR activation as an early adaptive response of KRAS-dependent CRC cells to KRAS silencing.
Document Type: Article
Other literature type
Language: English
ISSN: 2045-2322
DOI: 10.1038/s41598-025-94549-2
Access URL: https://pubmed.ncbi.nlm.nih.gov/40274922
https://doaj.org/article/2ac9e7d15b51492a9e9add08427f3361
Rights: CC BY NC ND
Accession Number: edsair.doi.dedup.....72f1c9264a15acfa1d6e08abd661e473
Database: OpenAIRE
Description
Abstract:Despite the development of mutant-selective KRAS inhibitors, colorectal cancer (CRC) responses remain limited, with stable disease and rapid recurrence being common outcomes. The molecular mechanisms enabling CRC cells to tolerate KRAS inhibition and ultimately develop resistance remain poorly understood. Here, we investigated early transcriptional and proteomic responses to KRAS silencing in 3D CRC cell line spheroid models, aiming to identify pathways associated with sensitivity or resistance to KRAS blockade. Cell lines were stratified into KRAS silencing-sensitive (HCT116 and SW480) and -resistant (LS174T and SW837) groups based on spheroid growth, cell cycle progression, and apoptosis induction. Transcriptional profiling revealed the unfolded protein response (UPR) and WNT/β-catenin signaling as pathways specifically upregulated in KRAS silencing-sensitive cells and downregulated in resistant cells. Proteomic analysis of membrane-enriched fractions further supported UPR deregulation, showing a pronounced downregulation of translation-related proteins in sensitive cells. Functional assays validated that the sensitive cell line HCT116 exhibits reduced protein aggregation and lower translational capacity upon KRAS knockdown, consistent with UPR activation. Pharmacological inhibition of IRE1α-mediated UPR signaling did not revert KRAS silencing-induced cell cycle arrest or apoptosis in this cell line. Collectively, our results highlight the UPR activation as an early adaptive response of KRAS-dependent CRC cells to KRAS silencing.
ISSN:20452322
DOI:10.1038/s41598-025-94549-2