TP53 Knockout Research Articles

Induction and loss of a TP53-dependent radioadaptive response in the human lymphoblastoid cell model TK6 and its abrogation by BCL2 over-expression

Purpose: To characterize the radioadaptive response in the human lymphoblastoid cell model TK6, and determine: (i) Whether repeated low dose exposures are more effective than single acute exposures in inducing resistance, (ii) the time-course for induction and loss of resistance following chronic exposures, and (iii) the effect of TP53 deletion or BCL2 over-expression on the induction of an adaptive response.Materials and methods: TK6, a human B-lymphoblastoid cell line, TK6-BCL2, a TK6 line that over-expresses BCL2 and is resistant to radiation-induced apoptosis, and NH32, a TP53 knockout of TK6 that is also resistant to apoptosis were studied. Cells were exposed to chronic, daily doses of 10 cGy given over 1 – 21 days before being challenged with 1 – 5 Gy exposures. Cell survival and chromatid break induction following high dose challenge were used to evaluate adaptive radiation responses.Results: Exposure to 10 cGy gamma rays induced resistance to killing and chromosome break induction in TK6 cells, but not in either TK6-BCL2 or NH32 cells. Resistance in TK6 was observed 4 h after exposure, and cells remained resistant for about 48 h. Maximal resistance was induced by a single 10 cGy dose. Repeated 10 cGy exposures had no additional effect on radiation sensitivity, except to maintain the induced radioresistance.Conclusion: An adaptive response is maximally and rapidly induced by a single low dose exposure in TK6 cells, and it has a limited lifespan. Induction of an adaptive response in TK6 cells can be abrogated by either TP53 loss or BCL2 over-expression. The characteristics of induced resistance in TK6 cells suggest that alterations in TP53-dependent apoptotic responses may be one mechanism for resistance.

Study of the gonocyte cell cycle in irradiated TP53 knockout mouse foetuses and newborns

Purpose : To investigate the role of p53 in gonocyte cell-cycle arrest in rodents with or without radiation exposure. Materials and methods : Pregnant p53 (+/-) mice, mated with p53 (-/-) males, were exposed to 137 Cs γ-rays at day 18 postcoitum (p.c.) at doses ranging from 1.5 Gy to 3 Gy. The gonocyte cell cycle was studied in p53 (-/-) male foetuses and newborns after BrdU incorporation and DAPI staining, and compared with those of p53 wild-type animals. Results : The proliferation of gonocytes in wild-type mice is normally arrested between day 16 p.c. and birth, a period when p53 is strongly expressed in gonocytes; p53 high expression is prolonged in all gonocytes after 3 Gy irradiation. In p53 (-/-) mice, this period of gonocyte cell-cycle arrest is not modified, compared with wild-type mice. It is also prolonged after a 3 Gy exposure. Conclusion : Two hypotheses are proposed. Either p53 is not involved in the control of gonocyte cell-cycle arrest in control and irradiated mice, or its role is redundant in this process.