ABSTRACT
Objective To investigate the effects of ionizing radiation on the expression of P21 protein in Jurkat cell line and p21 gene in thymocytes and splenocytes of mice.Methods Flow cytometry (FCM)was used to analyze the expression of P21 protein in Jurkat cells at 12 and 24 h after irradiation to 0,0.5,1.0,2.0,4.0,and 6.0 Gy.Real-time PCR was used to detect the expression of p21 gene in thymocytes and splenocytes of mice at4 and 24 h after irradiation to 0,0.5,1.0,2.0,4.0,and 6.0 Gy.Multi-staining was used to analyze the micronucleus rates of Rct in bone marrow.Results The expressions of P21 protein were increased in a dose-dependent manner during 0.5-4.0 Gy(t=-24.23--3.96,P<0.05),but decreased at 6.0 Gy at 12 and 24 h post-irradiation(t=-11.19,-14.50,P<0.05).The expressions of p2 1 gene in both thymocytes and splenocytes of mice were increased in dose-dependent manner in the range of 0-6.0 Gy(including 6.0 Gy)(t=-29.96-8.80,P<0.05),and reached to the peak at 6.0 Gy at 4 and 24 h post-irradiation(t=-11.84--3.42,P<0.05),except thymocytes at 4 h and 1.0 Gy post-irradiation(t=-3.42,P>0.05).Conclusions The expressions of P21 protein and p21 gene could be increased by X-ray irradiation.which shows good dosedependent manners in certain range of dose.
ABSTRACT
Objective To detect the expression of DNA damage response genes induced by radiation in human peripheral blood lymphocyte,and to explore the new biomarkers of radiation.Methods The human peripheral blood cells were irradiated to X-rays at different doses of 0,1,2,3,4,and 5 Gy.The quantitative real.time qPCR wag used to detect the expressions of cyclin-dependent kinase inhibitor l a gene(Cdknl a)and growth arrest and DNA damage inducible gene(Gadd45a)in lymphoeytes at 4 and 24 h post-irradiation,respectively.The method of CB mieronucleus was used to determine the change of micronucleus ratio.Results The expression of Cdknl a in peripheral blood lymphocytes wag increased significantly at 4 and 24 h post-irradiation to 0-5 Gy.reached the peak at 4 Gy and began to decrease at 5 Gy,which showed a dose-dependent manner(r=0.946,0.975,P<0.05).Similarly,the expression of Gadd45α in human peripheral blood lymphocytes was also increased significantly at 4 and 24 h post-irradiation to 0-5 Gy in a dose-dependent manner,while the expression of Gadd45a at 4 h wag higher than that at 24 h(r=0.936,0.797,P<0.05).The ratio of micronuclei wag increased significantly at 4 and 24 h post-irradiation to 0-5 Gy(r=0.990,0.984,P<0.05).Conciusions Cdknl a and Gadd45α expression could be increaged significandy at 4 and 24 h post-irradiation to 0-5 Gy,showing a good linear relationship.which might be candidate for radiation biological dosimeter.
ABSTRACT
Objective To observe the changes in expressions of spleen regulatory T cells (Tregs)and the related factor forkhead box protein-3 (Foxp3) after irradiation with different doses of X-ray in mice at different times,and to elaborate the effects of X-rays on regulatory T cells and Foxp3.Methods 112male ICR mice were randomly divided into 2 groups and irradiated by X-rays at the doses of 0.075 and 2 Gy,respectively.The mice were killed at 0,4,8,16,24,48,and 72 h post-irradiation and the spleens removed.Flow cytometry was used to detect the percentage of CD4 + CD25 + Treg and protein expression of (Foxp3),and RT-PCR was used to exmiamine the mRNA expression of Fox3.Results Compared with those before irradiation,the CD4 + CD25 + Treg positive rates began to increase and peaked at 8 h post-irradiation with 0.075 Gy at 8,16,24,72 h(t = 8.73,10.55,4.21,4.65 ,P ＜ 0.05) and 2 Gy at 8,16,48,72 h(t = 4.65,4.28,3.71,2.88,P ＜ 0.05),and then slightly decreased,but still remained at high levels.The mRNA protein levels of Fox3 did not change significantly after exposure to the dose of 0.075 Gy,but began to significantly increase at 8 h after exposure to the dose of 2 Gy.However,the Foxp3 protein level began to increase 4 h post-irradiation,peaked at 16 h,and then slightly decreased,but still ramained at high levels (t =2.59,3.37,3.70,3.20,P＜0.05).Conclusions The changes in expressions of Tregs and Foxp3 after high- and low-dose X-ray irradiation may be used to explain the differences in immune effects induced by ionizing radiation at different doses.