CAJ | 학술논문

目的采用实时定量PCR技术,检测人外周血淋巴细胞DNA损伤反应相关基因表达对X射线全身照射的反应,为探索新型辐射生物标志物奠定基础.方法以吸收剂量为0、1、2、3、4、5 Gy X射线照射正常人外周血,在照射后4和24 h,应用实时定量PCR法,对淋巴细胞细胞周期素依赖性蛋白激酶抑制物蛋白1a(Cdknla)、生长阻滞和DNA损伤基因45a(Gadd45α)基因的表达变化进行检测.应用胞质分裂阻滞微核法(CB微核法),检测淋巴细胞微核率变化.结果 Cdknla基因在人外周血淋巴细胞受到1～5 Gy照射后4和24 h,其相对表达量均较对照组显著性升高,至4 Gy达到峰值,5 Gy后不再继续增加.Cdknla基因表达与照射剂量呈线性相关(r=0.946、0.975,P<0.05).Gadd45ct基因在1～5 Gy照后4和24 h,其相对表达量均呈剂量依赖性升高,且照射后4 h的表达高于24 h(r=0.936、0.797,P<0.05).CB微核法中,在1～5 Gy X射线照射后4和24 h,各剂量组淋巴细胞微核率均显著增多,呈现良好的线性关系(r=0.990、0.984,P<0.05).结论辐射使Cdknla基因和Gadd45α基因表达上调,表现出较好的剂量线性关系,有可能成为研制新型辐射生物剂量计的候选基因.
목적 채용실시정량PCR기술,검측인외주혈림파세포DNA손상반응상관기인표체대X사선전신조사적반응,위탐색신형복사생물표지물전정기출.방법 이흡수제량위0、1、2、3、4、5 Gy X사선조사정상인외주혈,재조사후4화24 h,응용실시정량PCR법,대림파세포세포주기소의뢰성단백격매억제물단백1a(Cdknla)、생장조체화DNA손상기인45a(Gadd45α)기인적표체변화진행검측.응용포질분렬조체미핵법(CB미핵법),검측림파세포미핵솔변화.결과 Cdknla기인재인외주혈림파세포수도1～5 Gy조사후4화24 h,기상대표체량균교대조조현저성승고,지4 Gy체도봉치,5 Gy후불재계속증가.Cdknla기인표체여조사제량정선성상관(r=0.946、0.975,P<0.05).Gadd45ct기인재1～5 Gy조후4화24 h,기상대표체량균정제량의뢰성승고,차조사후4 h적표체고우24 h(r=0.936、0.797,P<0.05).CB미핵법중,재1～5 Gy X사선조사후4화24 h,각제량조림파세포미핵솔균현저증다,정현량호적선성관계(r=0.990、0.984,P<0.05).결론 복사사Cdknla기인화Gadd45α기인표체상조,표현출교호적제량선성관계,유가능성위연제신형복사생물제량계적후선기인.
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.