CAJ | 학술논문

目的研究低剂量X射线对人树突状细胞(DC)体外迁移能力影响并探讨其机制.方法分离人外周血单个核细胞(PBMC),以人GM-CSF和IL-4共同诱导分化为DC,于培养的第5天加入TNF-α促进成熟,在培养的第6天用X射线照射DC,受照剂量分别是0、0.05、0.1、0.2、0.5 Gy,照射后48 h收获DC;采用RT-PCR法及Western blot法分别检测CCR7 mRNA及蛋白表达水平;Transwell迁移实验法检测DC的体外迁移能力.结果与0 Gy相比,0.2和0.5 Gy照射后,CCR7 mRNA的相对表达量显著高于其他剂量(t=14.72、4.72,P＜0.05);0.2 Gy照射后,CCR7蛋白表达量高于其他剂量(t=4.46,P＜0.05),DC迁移能力显著高于其他剂量(t=2.95,P＜0.05);以抗CCR7单克隆抗体封闭CCR7蛋白活性,在接受同等剂量照射时,DC细胞迁移能力显著降低(t=4.63～8.96,P＜0.05).结论受到0.2 Gy X射线照射的DC,体外迁移能力显著增强,其机制可能与受照后DC表达CCR7 mRNA及蛋白水平升高有关.
목적 연구저제량X사선대인수돌상세포(DC)체외천이능력영향병탐토기궤제.방법 분리인외주혈단개핵세포(PBMC),이인GM-CSF화IL-4공동유도분화위DC,우배양적제5천가입TNF-α촉진성숙,재배양적제6천용X사선조사DC,수조제량분별시0、0.05、0.1、0.2、0.5 Gy,조사후48 h수획DC;채용RT-PCR법급Western blot법분별검측CCR7 mRNA급단백표체수평;Transwell천이실험법검측DC적체외천이능력.결과 여0 Gy상비,0.2화0.5 Gy조사후,CCR7 mRNA적상대표체량현저고우기타제량(t=14.72、4.72,P＜0.05);0.2 Gy조사후,CCR7단백표체량고우기타제량(t=4.46,P＜0.05),DC천이능력현저고우기타제량(t=2.95,P＜0.05);이항CCR7단극륭항체봉폐CCR7단백활성,재접수동등제량조사시,DC세포천이능력현저강저(t=4.63～8.96,P＜0.05).결론 수도0.2 Gy X사선조사적DC,체외천이능력현저증강,기궤제가능여수조후DC표체CCR7 mRNA급단백수평승고유관.
Objective To study the effect of low dose X-ray irradiation on the migration of human dendritic cells and its mechanism in vitro.Methods The human peripheral blood mononuclear cells (PBMC)were separated and treated with rhGM-CSF and rhIL-4 in order to differentiate them to dendritic cells(DC)in vitro.To enhance cell maturation,50 mg/L TNF-α was added into the medium at 5 d of culture.At 6 d of cell culture,DCs were radiated with X-rays of 0.05,0.1,0.2 and 0.5 Gy,respectively.At 8 d,the dendrite cells were collected for further analysis.The expressions of CC-Chemokine Receptor 7(CCR7)mRNA and its protein were detected by RT-PCR and Western blot,respectively.Transwell culture inserts were used to measure the amount of migrated cells.Results After 0.2 and 0.5 Gy radiation,the expression of CCR7 mRNA of DCs was remarkably increased(t=14.72,4.72,P＜0.05),but the expression of CCR7 protein in the DCs irradiated with 0.2 Gy was higher than that irradiated with other doses(t=4.46,P＜0.05),meanwhile the amount of migrated DCs was obviously increased(t=2.95,P＜O.05).Furthermore,DCs treated with anti-CCR7 monoclone antibody decreased the ability of radiation-induced migration(t=4.63-8.96,P＜0.05).Conclusions 0.2 Gy X-ray irradiation significantly can enhance the migration ability of DCs in vitro,which may be correlated with the increase of CCR7 expression.