HNSCC cell lines positive for HPV and p16 possess higher cellular radiosensitivity due to an impaired DSB repair capacity

When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16INK4a possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enha...

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Vydané v:	Radiotherapy and oncology Ročník 107; číslo 2; s. 242 - 246
Hlavní autori:	Rieckmann, Thorsten, Tribius, Silke, Grob, Tobias J., Meyer, Felix, Busch, Chia-Jung, Petersen, Cordula, Dikomey, Ekkehard, Kriegs, Malte
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	Ireland Elsevier Ireland Ltd 01.05.2013
Predmet:	Apoptosis Carcinoma, Squamous Cell - chemistry Carcinoma, Squamous Cell - genetics Carcinoma, Squamous Cell - radiotherapy Carcinoma, Squamous Cell - virology Cell Line, Tumor Cyclin-Dependent Kinase Inhibitor p16 DNA Breaks, Double-Stranded DNA Repair G1 Phase Head and Neck Neoplasms - chemistry Head and Neck Neoplasms - genetics Head and Neck Neoplasms - radiotherapy Head and Neck Neoplasms - virology Hematology, Oncology, and Palliative Medicine HNSCC HPV Humans Neoplasm Proteins - analysis p16INK4a Papillomaviridae - isolation & purification Radiation Tolerance Radiosensitivity Squamous Cell Carcinoma of Head and Neck Radiosensitivity p16INK4a HNSCC HPV p16 INK4a
ISSN:	0167-8140, 1879-0887, 1879-0887
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Abstract	When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16INK4a possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enhanced cellular radiosensitivity. The radiation response of five HPV/p16INK4a-positive and five HPV-negative cell lines was characterized with regard to cellular radiosensitivity by colony formation assay. Furthermore G1- and G2-arrest, apoptosis and residual DNA double-strand breaks (DSB) were analyzed by the colcemid-based G1-efflux assay, propidium iodide staining, the detection of PARP cleavage, the fluorescence-based detection of caspase activity and the immunofluorescence staining of γH2AX and 53BP1 foci. On average, the cellular radiosensitivity of the HNSCC cell lines positive for HPV and p16INK4a was higher as compared to the sensitivity of a panel of five HPV-negative HNSCC cell lines (SF3=0.2827 vs. 0.4455). The higher sensitivity does not result from increased apoptosis or the execution of a permanent G1-arrest, but is rather associated with both, elevated levels of residual DSBs and extensive G2-arrest. Increased cellular radiosensitivity due to compromised DNA repair capacity is likely to contribute to the improved outcome of patients with HPV/p16INK4a-positive tumors when treated by radiotherapy.
AbstractList	When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16INK4a possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enhanced cellular radiosensitivity. The radiation response of five HPV/p16INK4a-positive and five HPV-negative cell lines was characterized with regard to cellular radiosensitivity by colony formation assay. Furthermore G1- and G2-arrest, apoptosis and residual DNA double-strand breaks (DSB) were analyzed by the colcemid-based G1-efflux assay, propidium iodide staining, the detection of PARP cleavage, the fluorescence-based detection of caspase activity and the immunofluorescence staining of γH2AX and 53BP1 foci. On average, the cellular radiosensitivity of the HNSCC cell lines positive for HPV and p16INK4a was higher as compared to the sensitivity of a panel of five HPV-negative HNSCC cell lines (SF3=0.2827 vs. 0.4455). The higher sensitivity does not result from increased apoptosis or the execution of a permanent G1-arrest, but is rather associated with both, elevated levels of residual DSBs and extensive G2-arrest. Increased cellular radiosensitivity due to compromised DNA repair capacity is likely to contribute to the improved outcome of patients with HPV/p16INK4a-positive tumors when treated by radiotherapy. AbstractBackground and purposeWhen treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16 INK4a possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enhanced cellular radiosensitivity. Materials and methodsThe radiation response of five HPV/p16 INK4a-positive and five HPV-negative cell lines was characterized with regard to cellular radiosensitivity by colony formation assay. Furthermore G1- and G2-arrest, apoptosis and residual DNA double-strand breaks (DSB) were analyzed by the colcemid-based G1-efflux assay, propidium iodide staining, the detection of PARP cleavage, the fluorescence-based detection of caspase activity and the immunofluorescence staining of γH2AX and 53BP1 foci. ResultsOn average, the cellular radiosensitivity of the HNSCC cell lines positive for HPV and p16 INK4a was higher as compared to the sensitivity of a panel of five HPV-negative HNSCC cell lines (SF3 = 0.2827 vs. 0.4455). The higher sensitivity does not result from increased apoptosis or the execution of a permanent G1-arrest, but is rather associated with both, elevated levels of residual DSBs and extensive G2-arrest. ConclusionsIncreased cellular radiosensitivity due to compromised DNA repair capacity is likely to contribute to the improved outcome of patients with HPV/p16 INK4a-positive tumors when treated by radiotherapy. When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16(INK4a) possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enhanced cellular radiosensitivity.BACKGROUND AND PURPOSEWhen treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16(INK4a) possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enhanced cellular radiosensitivity.The radiation response of five HPV/p16(INK4a)-positive and five HPV-negative cell lines was characterized with regard to cellular radiosensitivity by colony formation assay. Furthermore G1- and G2-arrest, apoptosis and residual DNA double-strand breaks (DSB) were analyzed by the colcemid-based G1-efflux assay, propidium iodide staining, the detection of PARP cleavage, the fluorescence-based detection of caspase activity and the immunofluorescence staining of γH2AX and 53BP1 foci.MATERIALS AND METHODSThe radiation response of five HPV/p16(INK4a)-positive and five HPV-negative cell lines was characterized with regard to cellular radiosensitivity by colony formation assay. Furthermore G1- and G2-arrest, apoptosis and residual DNA double-strand breaks (DSB) were analyzed by the colcemid-based G1-efflux assay, propidium iodide staining, the detection of PARP cleavage, the fluorescence-based detection of caspase activity and the immunofluorescence staining of γH2AX and 53BP1 foci.On average, the cellular radiosensitivity of the HNSCC cell lines positive for HPV and p16(INK4a) was higher as compared to the sensitivity of a panel of five HPV-negative HNSCC cell lines (SF3=0.2827 vs. 0.4455). The higher sensitivity does not result from increased apoptosis or the execution of a permanent G1-arrest, but is rather associated with both, elevated levels of residual DSBs and extensive G2-arrest.RESULTSOn average, the cellular radiosensitivity of the HNSCC cell lines positive for HPV and p16(INK4a) was higher as compared to the sensitivity of a panel of five HPV-negative HNSCC cell lines (SF3=0.2827 vs. 0.4455). The higher sensitivity does not result from increased apoptosis or the execution of a permanent G1-arrest, but is rather associated with both, elevated levels of residual DSBs and extensive G2-arrest.Increased cellular radiosensitivity due to compromised DNA repair capacity is likely to contribute to the improved outcome of patients with HPV/p16(INK4a)-positive tumors when treated by radiotherapy.CONCLUSIONSIncreased cellular radiosensitivity due to compromised DNA repair capacity is likely to contribute to the improved outcome of patients with HPV/p16(INK4a)-positive tumors when treated by radiotherapy. When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16(INK4a) possess a clearly favorable prognosis as compared to those with HPV-negative HNSCC. The aim of this work was to study whether the better outcomes might be caused by an enhanced cellular radiosensitivity. The radiation response of five HPV/p16(INK4a)-positive and five HPV-negative cell lines was characterized with regard to cellular radiosensitivity by colony formation assay. Furthermore G1- and G2-arrest, apoptosis and residual DNA double-strand breaks (DSB) were analyzed by the colcemid-based G1-efflux assay, propidium iodide staining, the detection of PARP cleavage, the fluorescence-based detection of caspase activity and the immunofluorescence staining of γH2AX and 53BP1 foci. On average, the cellular radiosensitivity of the HNSCC cell lines positive for HPV and p16(INK4a) was higher as compared to the sensitivity of a panel of five HPV-negative HNSCC cell lines (SF3=0.2827 vs. 0.4455). The higher sensitivity does not result from increased apoptosis or the execution of a permanent G1-arrest, but is rather associated with both, elevated levels of residual DSBs and extensive G2-arrest. Increased cellular radiosensitivity due to compromised DNA repair capacity is likely to contribute to the improved outcome of patients with HPV/p16(INK4a)-positive tumors when treated by radiotherapy.
Author	Grob, Tobias J. Meyer, Felix Busch, Chia-Jung Rieckmann, Thorsten Tribius, Silke Kriegs, Malte Dikomey, Ekkehard Petersen, Cordula
Author_xml	– sequence: 1 givenname: Thorsten surname: Rieckmann fullname: Rieckmann, Thorsten email: t.rieckmann@uke.uni-hamburg.de organization: Laboratory of Radiobiology & Experimental Radiooncology, University Medical Center Hamburg Eppendorf, Germany – sequence: 2 givenname: Silke surname: Tribius fullname: Tribius, Silke organization: Department of Radiotherapy and Radiooncology, University Medical Center Hamburg Eppendorf, Germany – sequence: 3 givenname: Tobias J. surname: Grob fullname: Grob, Tobias J. organization: Department of Pathology, University Medical Center Hamburg Eppendorf, Germany – sequence: 4 givenname: Felix surname: Meyer fullname: Meyer, Felix organization: Laboratory of Radiobiology & Experimental Radiooncology, University Medical Center Hamburg Eppendorf, Germany – sequence: 5 givenname: Chia-Jung surname: Busch fullname: Busch, Chia-Jung organization: Department of Otolaryngology and Head and Neck Surgery, University Medical Center Hamburg Eppendorf, Hamburg, Germany – sequence: 6 givenname: Cordula surname: Petersen fullname: Petersen, Cordula organization: Department of Radiotherapy and Radiooncology, University Medical Center Hamburg Eppendorf, Germany – sequence: 7 givenname: Ekkehard surname: Dikomey fullname: Dikomey, Ekkehard organization: Laboratory of Radiobiology & Experimental Radiooncology, University Medical Center Hamburg Eppendorf, Germany – sequence: 8 givenname: Malte surname: Kriegs fullname: Kriegs, Malte organization: Laboratory of Radiobiology & Experimental Radiooncology, University Medical Center Hamburg Eppendorf, Germany
BackLink	https://www.ncbi.nlm.nih.gov/pubmed/23602369$$D View this record in MEDLINE/PubMed
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Snippet	When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16INK4a possess a clearly favorable... AbstractBackground and purposeWhen treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16 INK4a... When treated by radiotherapy, patients with squamous cell carcinomas of the head and neck (HNSCC) positive for HPV and p16(INK4a) possess a clearly favorable...
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SubjectTerms	Apoptosis Carcinoma, Squamous Cell - chemistry Carcinoma, Squamous Cell - genetics Carcinoma, Squamous Cell - radiotherapy Carcinoma, Squamous Cell - virology Cell Line, Tumor Cyclin-Dependent Kinase Inhibitor p16 DNA Breaks, Double-Stranded DNA Repair G1 Phase Head and Neck Neoplasms - chemistry Head and Neck Neoplasms - genetics Head and Neck Neoplasms - radiotherapy Head and Neck Neoplasms - virology Hematology, Oncology, and Palliative Medicine HNSCC HPV Humans Neoplasm Proteins - analysis p16INK4a Papillomaviridae - isolation & purification Radiation Tolerance Radiosensitivity Squamous Cell Carcinoma of Head and Neck
Title	HNSCC cell lines positive for HPV and p16 possess higher cellular radiosensitivity due to an impaired DSB repair capacity
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