Effect of radiation on Ewing tumour subpopulations characterized on a single-cell level: intracellular cytokine, immunophenotypic, DNA and apoptotic profile.
OBJECTIVE
Adhesion molecules, cytokines and their corresponding cell-surface receptors are involved in intercellular signalling pathways, radioresistance and metastasis-mediating mechanisms of malignant cells. The aim was the characterization of changes in the marker profile of Ewing tumour cell subpopulations under the influence of radiation.


METHODS
Three Ewing tumours were characterized in vitro and in vivo in a xenograft model before and after radiation by five-parameter flow cytometry. Antibodies directed against cell surface and intracellular antigens, apoptosis-associated markers and the DNA dye 7-aminoactinomycin D were used.


RESULTS
Tumour cell subpopulations were identified by expression of adhesion molecules and cytokine receptors, intracellular cytokines, apoptotic markers and DNA content. Heterogeneous changes of flow cytometric profile were identified on tumour cell subpopulations after radiation.


CONCLUSIONS
The changed profile of tumour cells under radiation might be associated with biological changes of tumour subpopulations in view of radioresistance and metastatic potential and might be useful to identify intercellular regulation mechanisms and to define parameters being predictive for a response to therapy.
University Hospital Münster, Department of Radiotherapy, Albert-Schweitzer-Str 33, D-48129 Münster, Germany. stkoene@uni-muenster.de
S
S
Könemann
T
T
Bölling
A
A
Schuck
J
J
Malath
A
A
Kolkmeyer
K
K
Horn
D
D
Riesenbeck
S
S
Hesselmann
R
R
Diallo
J
J
Vormoor
N A
NA
Willich
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D019002:CD56 Antigen / Q000096:biosynthesis; D002448:Cell Adhesion; D016207:Cytokines / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D003609:Dactinomycin / Q000031:analogs & derivatives* / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D012512:Sarcoma, Ewing / Q000473:pathology*; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation
D017304:Annexin A5; D019002:CD56 Antigen; D016207:Cytokines; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; D003609:Dactinomycin; C025942:7-aminoactinomycin D; D004247:DNA