Role of p53 mutations, protein function and DNA damage for the radiosensitivity of human tumour cells.
OBJECTIVE
The tumour suppressor protein p53 is considered to have an impact on the radiosensitivity of tumour cells. However, this concept does not easily translate to the tumour sensitivity in the clinics. The aim of the present study was to determine whether a functional or dysfunctional p53 is associated with a sensitive or resistant phenotype. It was further studied whether DNA damage might be an additive factor by which p53 has impact on cell survival.


METHODS
Nine human tumour cell lines were studied for p53 mutation by direct sequencing of exons 4-9. Regulation of p53 and p21(cip1/waf1) protein was assessed by immunoblotting and cell cycle effects by combining 5-bromodeoxyuridine incorporation and flow cytometry.


CONCLUSIONS
Three strains (RT112, Du145, SCC4451) were found to have a missense-mutation in the core domain and one did not express p53 at all (HeLa), presumably due to HPV18 infection. Immunoblots of these cells showed neither a regulated p53 nor p21 expression. The cells did not arrest in G1 phase after X-irradiation but did arrest in G2/M. All cells expressing wild-type protein (LNCaP, T47D-B8, MCF-7 and sublines BB and Bus) showed an intact p53 and p21 regulation and a modest arrest in both G1 and G2/M. Thus, in contrast to other studies, all tumour cells investigated showed either a typical p53wt or mutant (mut) pattern. Protein function was compared with cell survival and DNA damage, as assessed previously. p53 wild-type cells were on average 1.3-times (n.s.) more radiosensitive than mutant cells, but there was a considerable overlap between both groups. Further, the 1.3-fold enhanced resistance of cells lacking wild-type p53 was paralleled by a 1.3-fold lower number of induced double-strand breaks. The results suggest that p53 could have impact on chromatin compaction and thus effect DNA damage induction and radiosensitivity of tumour cells.
Böhnke
A
A
University Hospital of Hamburg-Eppendorf, Department of Radiotherapy and Radiation Oncologyk Martinistr. 52k D-20246 Hamburgk Germany.
Westphal
F
F
Schmidt
A
A
El-Awady
R A
RA
Dahm-Daphi
J
J
D000222:Adaptation, Physiological; D002453:Cell Cycle / Q000528:radiation effects; D045744:Cell Line, Tumor / Q000201:enzymology / Q000528:radiation effects; D003001:Cloning, Molecular; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009369:Neoplasms / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D011829:Radiation Dosage; D011836:Radiation Tolerance / Q000235:genetics*; D013329:Structure-Activity Relationship; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D016159:Tumor Suppressor Protein p53; D004247:DNA