Publicação
Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells
| Resumo: | 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. |
|---|---|
| Autores principais: | Martins, Flávia |
| Outros Autores: | Machado, Ana L.; Carvalho, Joana; Almeida, Catarina R.; Beck, Hans C.; Carvalho, Ana S.; Backman, Vadim; Matthiesen, Rune; Matthiesen, Rune; Velho, Sérgia |
| Assunto: | General SDG 3 - Good Health and Well-being |
| Ano: | 2025 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| _version_ | 1868983064494866432 |
|---|---|
| author | Martins, Flávia |
| author2 | Machado, Ana L. Carvalho, Joana Almeida, Catarina R. Beck, Hans C. Carvalho, Ana S. Backman, Vadim Matthiesen, Rune Matthiesen, Rune Velho, Sérgia |
| author2_role | author author author author author author author author author |
| author_facet | Martins, Flávia Machado, Ana L. Carvalho, Joana Almeida, Catarina R. Beck, Hans C. Carvalho, Ana S. Backman, Vadim Matthiesen, Rune Matthiesen, Rune Velho, Sérgia |
| author_role | author |
| contributor_name_str_mv | NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM) iNOVA4Health - pólo NMS Nature Publishing Group RUN |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Martins, Flávia\"},{\"Person.name\":\"Machado, Ana L.\"},{\"Person.name\":\"Carvalho, Joana\"},{\"Person.name\":\"Almeida, Catarina R.\"},{\"Person.name\":\"Beck, Hans C.\"},{\"Person.name\":\"Carvalho, Ana S.\"},{\"Person.name\":\"Backman, Vadim\"},{\"Person.name\":\"Matthiesen, Rune\"},{\"Person.name\":\"Matthiesen, Rune\",\"Person.identifier.orcid\":\"0000-0002-6353-2616\"},{\"Person.name\":\"Velho, Sérgia\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM) iNOVA4Health - pólo NMS Nature Publishing Group RUN |
| datacite.creators.creator.creatorName.fl_str_mv | Martins, Flávia Machado, Ana L. Carvalho, Joana Almeida, Catarina R. Beck, Hans C. Carvalho, Ana S. Backman, Vadim Matthiesen, Rune Matthiesen, Rune Velho, Sérgia |
| datacite.date.Accepted.fl_str_mv | 2025-04-01T00:00:00Z |
| datacite.date.available.fl_str_mv | 2025-05-07T21:35:28Z |
| datacite.date.embargoed.fl_str_mv | 2025-05-07T21:35:28Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | General SDG 3 - Good Health and Well-being |
| datacite.titles.title.fl_str_mv | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| dc.contributor.none.fl_str_mv | NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM) iNOVA4Health - pólo NMS Nature Publishing Group RUN |
| dc.creator.none.fl_str_mv | Martins, Flávia Machado, Ana L. Carvalho, Joana Almeida, Catarina R. Beck, Hans C. Carvalho, Ana S. Backman, Vadim Matthiesen, Rune Matthiesen, Rune Velho, Sérgia |
| dc.date.Accepted.fl_str_mv | 2025-04-01T00:00:00Z |
| dc.date.available.fl_str_mv | 2025-05-07T21:35:28Z |
| dc.date.embargoed.fl_str_mv | 2025-05-07T21:35:28Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10362/182833 |
| dc.language.none.fl_str_mv | eng |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | General SDG 3 - Good Health and Well-being |
| dc.title.fl_str_mv | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_6501 |
| 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. |
| dirty | 0 |
| eu_rights_str_mv | openAccess |
| format | article |
| fulltext.url.fl_str_mv | https://run.unl.pt/bitstreams/9bac49d9-8ad9-447a-9b1e-1dd29d94a151/download |
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| identifier.url.fl_str_mv | http://hdl.handle.net/10362/182833 |
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| language | eng |
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| oai_identifier_str | oai:run.unl.pt:10362/182833 |
| organization_str_mv | urn:organizationAcronym:unl |
| person_str_mv | Martins, Flávia Machado, Ana L. Carvalho, Joana Almeida, Catarina R. Beck, Hans C. Carvalho, Ana S. Backman, Vadim Matthiesen, Rune Matthiesen, Rune Matthiesen, Rune https://www.ciencia-id.pt/BB1A-A606-EE77 BB1A-A606-EE77 http://orcid.org/0000-0002-6353-2616 0000-0002-6353-2616 Velho, Sérgia |
| publishDate | 2025 |
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| reponame_str | Repositório Institucional da UNL |
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| spelling | engenDespite 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.application/pdfenDifferential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cellsMartins, FláviaMachado, Ana L.Carvalho, JoanaAlmeida, Catarina R.Beck, Hans C.Carvalho, Ana S.Backman, VadimMatthiesen, RunePersonalMatthiesen, RuneDSpacehttp://dspace.org/items/67181d3c-b83e-4151-9a74-0ea14aeb7f05DSpacehttp://dspace.org/items/67181d3c-b83e-4151-9a74-0ea14aeb7f05MatthiesenRuneCiência IDhttps://www.ciencia-id.ptBB1A-A606-EE77ORCIDhttp://orcid.org0000-0002-6353-2616Scopus Author IDhttps://www.scopus.com9741897800Velho, SérgiaNOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)iNOVA4Health - pólo NMSNature Publishing GroupHostingInstitutionOrganizationalRUNe-mailmailto:run@unl.ptrun@unl.ptISSNIsPartOf2045-2322URNIsPartOfPURE: 115798878URNIsPartOfPURE UUID: 8c38c8a4-12e2-4ffa-a389-c748f3daed07URNIsPartOfScopus: 105003450752DOIIsPartOf10.1038/s41598-025-94549-22025-05-07T21:35:28Z2025-042025-04-01T00:00:00ZHandlehttp://hdl.handle.net/10362/182833http://purl.org/coar/access_right/c_abf2open accessGeneralSDG 3 - Good Health and Well-being8378194 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://run.unl.pt/bitstreams/9bac49d9-8ad9-447a-9b1e-1dd29d94a151/download |
| spellingShingle | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells Martins, Flávia General SDG 3 - Good Health and Well-being |
| status | SINGLETON |
| subject.fl_str_mv | General SDG 3 - Good Health and Well-being |
| title | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| title_full | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| title_fullStr | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| title_full_unstemmed | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| title_short | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| title_sort | Differential unfolded protein response regulation in KRAS silencing sensitive and innately resistant colorectal cancer cells |
| topic | General SDG 3 - Good Health and Well-being |
| topic_facet | General SDG 3 - Good Health and Well-being |
| url | http://hdl.handle.net/10362/182833 |
| visible | 1 |