CAJ | 학술논문

目的探讨正常人淋巴母细胞AHH1电离辐射作用后PIG3基因mRNA表达变化的剂量-效应规律,建立一种基于基因表达变化的新型辐射生物剂量计的可能性.方法激光共聚焦检测DNA双链断裂分子标记γ-H2AX集簇点,1、2、4、6、8和10 Gy 60Coγ射线照射AHHI细胞后4、10和24h收集细胞,应用实时荧光定量PCR技术对PIG3基因mRNA表达水平进行相对定量检测.结果 γ射线照射后30 min检测发现PIG3蛋白与γ-H2AX在细胞核内有部分共定位,形成集簇点(foci),表明其参与电离辐射致DNA双链断裂损伤信号识别反应.γ射线诱导PIG3基因mRNA表达水平随时间推移不断提高,持续时间可达24 h.PIG3基因mRNA表达水平具有显著的剂量依赖性,其表达丰度变化的剂量依赖范围在照射后4h为0～6Gy、照射后10和24 h均为0～10 Gy.结论 PIG3基因参与DNA双链断裂损伤信号识别反应,其mRNA的辐射诱导表达具有良好的剂量-效应关系,具有发展成为新的放射生物剂量计的价值.
목적 탐토정상인림파모세포AHH1전리복사작용후PIG3기인mRNA표체변화적제량-효응규률,건립일충기우기인표체변화적신형복사생물제량계적가능성.방법 격광공취초검측DNA쌍련단렬분자표기γ-H2AX집족점,1、2、4、6、8화10 Gy 60Coγ사선조사AHHI세포후4、10화24h수집세포,응용실시형광정량PCR기술대PIG3기인mRNA표체수평진행상대정량검측.결과 γ사선조사후30 min검측발현PIG3단백여γ-H2AX재세포핵내유부분공정위,형성집족점(foci),표명기삼여전리복사치DNA쌍련단렬손상신호식별반응.γ사선유도PIG3기인mRNA표체수평수시간추이불단제고,지속시간가체24 h.PIG3기인mRNA표체수평구유현저적제량의뢰성,기표체봉도변화적제량의뢰범위재조사후4h위0～6Gy、조사후10화24 h균위0～10 Gy.결론 PIG3기인삼여DNA쌍련단렬손상신호식별반응,기mRNA적복사유도표체구유량호적제량-효응관계,구유발전성위신적방사생물제량계적개치.
Objective To investigate the dose-response pattern on the inducible expression of PIG3 mRNA in normal human lymphoblastoid AHHI cells by 60Co γ-rays,and its possibility for developing novel radiation biodosimeter.Methods Laser confocal fluorescent microscopy was used to detect the γ-H2AX foci,a biomarker of DNA double-strand break.After irradiation with 0,1,2,4,6,8 and 10 Gy of 60Coγ- rays,AHH-1 cells were harvested at 4,10 and 24 h post-irradiation,and subjected to total RNA extraction and detection of PIG3 mRNA expression by quantitative real-time PCR.Results PIG3 protein foci were formed in the nuclei at 30 min after irradiation,and a part of these PIG3 foci were colocalized with γH2AX foci.After irradiation,PIG3 mRNA level was enhanced with the increased time of postirradiation,and remained an increased level at least till 24 h.The radiation-inducible expression of PIG3 mRNA was demonstrated in a dose-dependent manner.The dose-dependent range at 4 h post-irradiation was 0 - 6 Gy,and at 10 h and 24 h was 0 - 10 Gy.Conclusions PIG3 involves in the cellular response to DNA double-strand break.The dose-dependent inducible response of PIG3 mRNA expression might provide a valuable candidate for developing a novel radiation biodosimeter.