CAJ | 학술논문

目的探讨在石英刺激的人胚肺成纤维细胞(human embryo lung fibroblasts,HELF)中Ku80蛋白表达的变化及Ku80/p53通路在石英诱导的细胞周期改变中的作用.方法 RNAi技术抑制Ku80蛋白表达,流式细胞技术检测细胞周期,免疫印迹技术检测Ku80、p53、p21蛋白的表达及p53-ser15磷酸化水平,并用Image-Pro plus 6.0软件对条带光强度进行半定量分析.结果 Ku80蛋白表达与石英刺激呈剂量-反应和时间-反应关系;石英刺激阴性对照H-NC细胞,G1期细胞所占比例从89.28%±2.19%下降到68.93%±3.79%;抑制Ku80蛋白表达的HELF细胞(H-Ku80)的G1期细胞所占比例进一步减少,从85.16%±3.73%下降到59.92%±3131%,差异均有统计学意义(P<0.05);抑制Ku80蛋白表达后,石英引起的p53、p21蛋白及p53-ser15磷酸化水平增高受抑制.结论 Ku80对p53、p21蛋白表达及p53-ser15磷酸化水平起调节作用,Ku80/p53通路可能参与了石英诱导的细胞周期改变.
목적 탐토재석영자격적인배폐성섬유세포(human embryo lung fibroblasts,HELF)중Ku80단백표체적변화급Ku80/p53통로재석영유도적세포주기개변중적작용.방법 RNAi기술억제Ku80단백표체,류식세포기술검측세포주기,면역인적기술검측Ku80、p53、p21단백적표체급p53-ser15린산화수평,병용Image-Pro plus 6.0연건대조대광강도진행반정량분석.결과 Ku80단백표체여석영자격정제량-반응화시간-반응관계;석영자격음성대조H-NC세포,G1기세포소점비례종89.28%±2.19%하강도68.93%±3.79%;억제Ku80단백표체적HELF세포(H-Ku80)적G1기세포소점비례진일보감소,종85.16%±3.73%하강도59.92%±3131%,차이균유통계학의의(P<0.05);억제Ku80단백표체후,석영인기적p53、p21단백급p53-ser15린산화수평증고수억제.결론 Ku80대p53、p21단백표체급p53-ser15린산화수평기조절작용,Ku80/p53통로가능삼여료석영유도적세포주기개변.
Objective To study the roles of Ku80/p53 pathway in silica-induced cell cycle changes in human embryo lung fibroblasts (HELF). Methods Ku80 siRNA expression vectors were transfected into HELF by lipofectamine. Flow cytometry was used to detect the distributions of cell cycle and western blot assay was used to determine the expression level of Ku80, p53 and p21 proteins or the phosphorylation levels of p53-serl5 after cells were exposed to silica. Results The expression levels of Ku80 protein increased in concentration-dependent and time-dependent manners after cells were exposed to silica. The proportion of G1 phases in H-NC cells (controls) decreased from 89.28%±2.19% to 68.93%±3.79% after exposure to silica, and the proportion of G1 phases in HELF cells (H-Ku80) decreased from 85.16%±3.73% to 59.92%±3.31% after exposure to silica (P<0.05). The expression levels of Ku80, p53 protecns or p21 proteins or phosphorylation level of p53-serl5 were obviously suppressed in H-Ku80, as compared with H-NC. Conclusion Ku80/p53 pathway plays a role in the cell cycle charges induced by silica in human embryo lung fibroblasts.