Integrative analysis of the microRNA-mRNA response to radiochemotherapy in primary head and neck squamous cell carcinoma cells.
BACKGROUND
Head and neck squamous cell carcinoma (HNSCC) is a very heterogeneous disease resulting in huge differences in the treatment response. New individualized therapy strategies including molecular targeting might help to improve treatment success. In order to identify potential targets, we developed a HNSCC radiochemotherapy cell culture model of primary HNSCC cells derived from two different patients (HN1957 and HN2092) and applied an integrative microRNA (miRNA) and mRNA analysis in order to gain information on the biological networks and processes of the cellular therapy response. We further identified potential target genes of four therapy-responsive miRNAs detected previously in the circulation of HNSCC patients by pathway enrichment analysis.


RESULTS
The two primary cell cultures differ in global copy number alterations and P53 mutational status, thus reflecting heterogeneity of HNSCC. However, they also share many copy number alterations and chromosomal rearrangements as well as deregulated therapy-responsive miRNAs and mRNAs. Accordingly, six common therapy-responsive pathways (direct P53 effectors, apoptotic execution phase, DNA damage/telomere stress induced senescence, cholesterol biosynthesis, unfolded protein response, dissolution of fibrin clot) were identified in both cell cultures based on deregulated mRNAs. However, inflammatory pathways represented an important part of the treatment response only in HN1957, pointing to differences in the treatment responses of the two primary cultures. Focused analysis of target genes of four therapy-responsive circulating miRNAs, identified in a previous study on HNSCC patients, revealed a major impact on the pathways direct P53 effectors, the E2F transcription factor network and pathways in cancer (mainly represented by the PTEN/AKT signaling pathway).


CONCLUSIONS
The integrative analysis combining miRNA expression, mRNA expression and the related cellular pathways revealed that the majority of radiochemotherapy-responsive pathways in primary HNSCC cells are related to cell cycle, proliferation, cell death and stress response (including inflammation). Despite the heterogeneity of HNSCC, the two primary cell cultures exhibited strong similarities in the treatment response. The findings of our study suggest potential therapeutic targets in the E2F transcription factor network and the PTEN/AKT signaling pathway.
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. isolde.summerer@helmholtz-muenchen.de.
Isolde
I
Summerer
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. julia.hess@helmholtz-muenchen.de.|Clinical Cooperation Group 'Personalized Radiotherapy of Head and Neck Cancer', Helmholtz Center Munich, Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany. julia.hess@helmholtz-muenchen.de.
Julia
J
Hess
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. adriana.pitea@helmholtz-muenchen.de.
Adriana
A
Pitea
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. unger@helmholtz-muenchen.de.|Clinical Cooperation Group 'Personalized Radiotherapy of Head and Neck Cancer', Helmholtz Center Munich, Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany. unger@helmholtz-muenchen.de.
Kristian
K
Unger
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. ludwig.hieber@helmholtz-muenchen.de.|Clinical Cooperation Group 'Personalized Radiotherapy of Head and Neck Cancer', Helmholtz Center Munich, Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany. ludwig.hieber@helmholtz-muenchen.de.
Ludwig
L
Hieber
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. martin.selmansberger@helmholtz-muenchen.de.
Martin
M
Selmansberger
Clinical Cooperation Group 'Personalized Radiotherapy of Head and Neck Cancer', Helmholtz Center Munich, Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany. kirsten.lauber@med.uni-muenchen.de.|Department of Radiation Oncology, University of Munich, Marchioninistr. 15, 81377, Munich, Germany. kirsten.lauber@med.uni-muenchen.de.
Kirsten
K
Lauber
Research Unit Radiation Cytogenetics, Helmholtz Center Munich, Ingolstaedter Landstr.1, 85764, Neuherberg, Germany. zitzelsberger@helmholtz-muenchen.de.|Clinical Cooperation Group 'Personalized Radiotherapy of Head and Neck Cancer', Helmholtz Center Munich, Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany. zitzelsberger@helmholtz-muenchen.de.
Horst
H
Zitzelsberger
D000368:Aged; D000369:Aged, 80 and over; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000628:therapy*; D045744:Cell Line, Tumor; D059248:Chemoradiotherapy; D016000:Cluster Analysis; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D006258:Head and Neck Neoplasms / Q000235:genetics* / Q000628:therapy*; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000235:genetics; D000077195:Squamous Cell Carcinoma of Head and Neck
D035683:MicroRNAs; D012333:RNA, Messenger