Increased G2 chromosomal radiosensitivity in cancer patients: the role of cdk1/cyclin-B activity level in the mechanisms involved.
OBJECTIVE
To test the hypothesis that deficient DNA repair as measured by increased G2 chromosomal radiosensitivity results from up-regulation of cdk1/cyclinB and cell cycle control mechanisms during the G2 to M transition.


METHODS
A total of 185 cancer patients and 25 normal individuals were tested for G2 chromosomal radiosensitivity. The chromatid breaks were analysed in metaphase using the G2 assay or directly in G0 and G2 phase using premature chromosome condensation (PCC). The activity of cdk1/cyclinB, a key regulator of the G2 to M-phase transition, was measured by histone H1 kinase activity and correlated with the development of chromatid breaks after irradiation of cell lines in vitro.


RESULTS
Based on the G2 assay, cancer patients on average showed increased chromosomal radiosensitivity above controls. When the analysis was carried out directly in G0 or G2 lymphocytes using PCC, no differences in the induction of chromosomal damage and its repair were observed between G2 assay-sensitive and G2-normal donors. Using the G2 assay to test G2 radiosensitivity in various cell lines, it was found that the higher the cdk1/cyclinB activity level of the cell line tested, the higher the yield of chromatid breaks scored. Furthermore, when mitotic cells from these cell lines were used for PCC induction in irradiated G2 lymphocytes it was observed that the higher the cdk1/cyclinB activity level of mitotic cells used, the higher was the induced yield of chromatid breaks.


CONCLUSIONS
The cdk1/cyclin-B activity levels during the G2 to M transition impair DNA repair processes and play a major role in the yield of chromatid breaks induced after G2-irradiation. Regulation of cdk1/cyclinB complex activity rather than deficient repair enzymes of DNA damage may underlie the mechanisms of G2 radiosensitivity.
Terzoudi
G I
GI
Institute of Nuclear Technology and Radiation Protection, National Centre for Scientific Research Demokritos, Athens, Greece.
Jung
T
T
Hain
J
J
Vrouvas
J
J
Margaritis
K
K
Donta-Bakoyianni
C
C
Makropoulos
V
V
Angelakis
P
P
Pantelias
G E
GE
D016203:CDC2 Protein Kinase / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D002875:Chromosomes / Q000528:radiation effects*; D019926:Cyclin B / Q000378:metabolism / Q000502:physiology*; D004249:DNA Damage / Q000528:radiation effects; D004260:DNA Repair / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D016195:G2 Phase / Q000235:genetics*; D020022:Genetic Predisposition to Disease; D006801:Humans; D008214:Lymphocytes / Q000528:radiation effects; D008938:Mitosis / Q000528:radiation effects; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism / Q000532:radiotherapy*; D013997:Time Factors; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
D019926:Cyclin B; D016203:CDC2 Protein Kinase