Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells

Girolamo Spagnoletti, Valeria Li Bergolis, Annamaria Piscazzi, Francesca Giannelli, Valentina Condelli, Lorenza Sisinni, Giuseppe Bove, Giovanni Storto, Matteo Landriscina

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)1010428318770957
JournalTumor Biology
Volume40
Issue number4
DOIs
Publication statusPublished - Apr 2018

Fingerprint

CDC2 Protein Kinase
Radiation Tolerance
Colorectal Neoplasms
HT29 Cells
Radiation
Carcinoma
Colon
HCT116 Cells
Cell Line
Standard of Care
Rectal Neoplasms
Ionizing Radiation
Fluorouracil
Quality Control
Mitochondria
Up-Regulation
Biomarkers

Keywords

  • CDC2 Protein Kinase/antagonists & inhibitors
  • Cell Line, Tumor
  • Chemoradiotherapy/methods
  • Colorectal Neoplasms/pathology
  • Fluorouracil/pharmacology
  • HCT116 Cells
  • HSP90 Heat-Shock Proteins/antagonists & inhibitors
  • HT29 Cells
  • Humans
  • Mitochondria/metabolism
  • Proto-Oncogene Proteins B-raf/antagonists & inhibitors
  • Radiation Tolerance/drug effects
  • Radiation, Ionizing
  • Radiation-Sensitizing Agents/pharmacology

Cite this

Spagnoletti, G., Li Bergolis, V., Piscazzi, A., Giannelli, F., Condelli, V., Sisinni, L., ... Landriscina, M. (2018). Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells. Tumor Biology, 40(4), 1010428318770957. https://doi.org/10.1177/1010428318770957

Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells. / Spagnoletti, Girolamo; Li Bergolis, Valeria; Piscazzi, Annamaria; Giannelli, Francesca; Condelli, Valentina; Sisinni, Lorenza; Bove, Giuseppe; Storto, Giovanni; Landriscina, Matteo.

In: Tumor Biology, Vol. 40, No. 4, 04.2018, p. 1010428318770957.

Research output: Contribution to journalArticle

Spagnoletti, G, Li Bergolis, V, Piscazzi, A, Giannelli, F, Condelli, V, Sisinni, L, Bove, G, Storto, G & Landriscina, M 2018, 'Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells', Tumor Biology, vol. 40, no. 4, pp. 1010428318770957. https://doi.org/10.1177/1010428318770957
Spagnoletti G, Li Bergolis V, Piscazzi A, Giannelli F, Condelli V, Sisinni L et al. Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells. Tumor Biology. 2018 Apr;40(4):1010428318770957. https://doi.org/10.1177/1010428318770957
Spagnoletti, Girolamo ; Li Bergolis, Valeria ; Piscazzi, Annamaria ; Giannelli, Francesca ; Condelli, Valentina ; Sisinni, Lorenza ; Bove, Giuseppe ; Storto, Giovanni ; Landriscina, Matteo. / Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells. In: Tumor Biology. 2018 ; Vol. 40, No. 4. pp. 1010428318770957.
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abstract = "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.",
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T1 - Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells

AU - Spagnoletti, Girolamo

AU - Li Bergolis, Valeria

AU - Piscazzi, Annamaria

AU - Giannelli, Francesca

AU - Condelli, Valentina

AU - Sisinni, Lorenza

AU - Bove, Giuseppe

AU - Storto, Giovanni

AU - Landriscina, Matteo

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N2 - 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.

AB - 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.

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KW - HSP90 Heat-Shock Proteins/antagonists & inhibitors

KW - HT29 Cells

KW - Humans

KW - Mitochondria/metabolism

KW - Proto-Oncogene Proteins B-raf/antagonists & inhibitors

KW - Radiation Tolerance/drug effects

KW - Radiation, Ionizing

KW - Radiation-Sensitizing Agents/pharmacology

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DO - 10.1177/1010428318770957

M3 - Article

C2 - 29663854

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JO - Tumor Biology

JF - Tumor Biology

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