Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells
Objectives: Preoperative chemoradiation is currently the standard of care in locally advanced rectal carcinoma, even though a subset of rectal tumors does not achieve major clinically meaningful responses upon neoadjuvant chemoradiation. At present, no molecular biomarkers are available to predict r...
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2018-04-01
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| Series: | Tumor Biology |
| Online Access: | https://doi.org/10.1177/1010428318770957 |
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| author | Girolamo Spagnoletti Valeria Li Bergolis Annamaria Piscazzi Francesca Giannelli Valentina Condelli Lorenza Sisinni Giuseppe Bove Giovanni Storto Matteo Landriscina |
| author_facet | Girolamo Spagnoletti Valeria Li Bergolis Annamaria Piscazzi Francesca Giannelli Valentina Condelli Lorenza Sisinni Giuseppe Bove Giovanni Storto Matteo Landriscina |
| author_sort | Girolamo Spagnoletti |
| collection | DOAJ |
| description | Objectives: Preoperative chemoradiation is currently the standard of care in locally advanced rectal carcinoma, even though a subset of rectal tumors does not achieve major clinically meaningful responses upon neoadjuvant chemoradiation. At present, no molecular biomarkers are available to predict response to neoadjuvant chemoradiation and select resistant tumors willing more intense therapeutic strategies. Thus, BRAF mutational status was investigated for its role in favoring resistance to radiation in colorectal carcinoma cell lines and cyclin-dependent kinase 1 as a target to improve radiosensitivity in BRAF V600E colorectal tumor cells. Methods: Colony-forming assay and apoptotic rates were evaluated to compare the sensitivity of different colon carcinoma cell lines to ionizing radiation and their radiosensitivity upon exposure to BRAF and/or cyclin-dependent kinase 1 inhibitory/silencing strategies. Cyclin-dependent kinase 1 expression/subcellular distribution was studied by immunoblot analysis. Results: Colon carcinoma BRAF V600E HT29 cells exhibited poor response to radiation compared to BRAF wild-type COLO320 and HCT116 cells. Interestingly, neither radiosensitizing doses of 5-fluoruracil nor BRAF inhibition/silencing significantly improved radiosensitivity in HT29 cells. Of note, poor response to radiation correlated with upregulation/relocation of cyclin-dependent kinase 1 in mitochondria. Consistently, cyclin-dependent kinase 1 inhibition/silencing as well as its targeting, through inhibition of HSP90 quality control pathway, significantly inhibited the clonogenic ability and increased apoptotic rates in HT29 cells upon exposure to radiation. Conclusion: These data suggest that BRAF V600E colorectal carcinoma cells are poorly responsive to radiation, and cyclin-dependent kinase 1 represents a target to improve radiosensitivity in BRAF V600E colorectal tumor cells. |
| format | Article |
| id | doaj-art-233c916fc10e47e5b1426dac20e99b80 |
| institution | DOAJ |
| issn | 1423-0380 |
| language | English |
| publishDate | 2018-04-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Tumor Biology |
| spelling | doaj-art-233c916fc10e47e5b1426dac20e99b802025-08-20T03:15:13ZengSAGE PublishingTumor Biology1423-03802018-04-014010.1177/1010428318770957Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cellsGirolamo Spagnoletti0Valeria Li Bergolis1Annamaria Piscazzi2Francesca Giannelli3Valentina Condelli4Lorenza Sisinni5Giuseppe Bove6Giovanni Storto7Matteo Landriscina8Radiotherapy Unit, Ospedali Riuniti, Foggia, ItalyMedical Oncology Unit, Department of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyMedical Oncology Unit, Department of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyMedical Oncology Unit, Department of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyLaboratory of Pre-Clinical and Translational Research, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture, Pz, ItalyLaboratory of Pre-Clinical and Translational Research, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture, Pz, ItalyRadiotherapy Unit, Ospedali Riuniti, Foggia, ItalyNuclear Medicine Unit, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture, Pz, ItalyLaboratory of Pre-Clinical and Translational Research, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture, Pz, ItalyObjectives: Preoperative chemoradiation is currently the standard of care in locally advanced rectal carcinoma, even though a subset of rectal tumors does not achieve major clinically meaningful responses upon neoadjuvant chemoradiation. At present, no molecular biomarkers are available to predict response to neoadjuvant chemoradiation and select resistant tumors willing more intense therapeutic strategies. Thus, BRAF mutational status was investigated for its role in favoring resistance to radiation in colorectal carcinoma cell lines and cyclin-dependent kinase 1 as a target to improve radiosensitivity in BRAF V600E colorectal tumor cells. Methods: Colony-forming assay and apoptotic rates were evaluated to compare the sensitivity of different colon carcinoma cell lines to ionizing radiation and their radiosensitivity upon exposure to BRAF and/or cyclin-dependent kinase 1 inhibitory/silencing strategies. Cyclin-dependent kinase 1 expression/subcellular distribution was studied by immunoblot analysis. Results: Colon carcinoma BRAF V600E HT29 cells exhibited poor response to radiation compared to BRAF wild-type COLO320 and HCT116 cells. Interestingly, neither radiosensitizing doses of 5-fluoruracil nor BRAF inhibition/silencing significantly improved radiosensitivity in HT29 cells. Of note, poor response to radiation correlated with upregulation/relocation of cyclin-dependent kinase 1 in mitochondria. Consistently, cyclin-dependent kinase 1 inhibition/silencing as well as its targeting, through inhibition of HSP90 quality control pathway, significantly inhibited the clonogenic ability and increased apoptotic rates in HT29 cells upon exposure to radiation. Conclusion: These data suggest that BRAF V600E colorectal carcinoma cells are poorly responsive to radiation, and cyclin-dependent kinase 1 represents a target to improve radiosensitivity in BRAF V600E colorectal tumor cells.https://doi.org/10.1177/1010428318770957 |
| spellingShingle | Girolamo Spagnoletti Valeria Li Bergolis Annamaria Piscazzi Francesca Giannelli Valentina Condelli Lorenza Sisinni Giuseppe Bove Giovanni Storto Matteo Landriscina Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells Tumor Biology |
| title | Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells |
| title_full | Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells |
| title_fullStr | Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells |
| title_full_unstemmed | Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells |
| title_short | Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells |
| title_sort | cyclin dependent kinase 1 targeting improves sensitivity to radiation in braf v600e colorectal carcinoma cells |
| url | https://doi.org/10.1177/1010428318770957 |
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