人肺腺癌细胞A549放射治疗前后Dystrobrevin蛋白表达的变化

Analysis of the Changes of Dystrobrevin Expression in Human Lung Adenocarcinoma A549 Cells Before and After Radiotherapy

目的射线诱导肺腺癌A549细胞凋亡,于凋亡高峰时检测放疗前后dystrobrevin的表达,探讨其与放射剂量及凋亡的关系。方法人肺腺癌A549细胞株培养24h后用10GyX线照射,分别培养6h、14h、23h、40h,应用Hoechest33258荧光染色在荧光显微镜下观察细胞形态变化——细胞的凋亡,并计算凋亡率。采用Westernblot检测0Gy、6Gy、10Gy照射A549细胞14h后dystrobrevin蛋白表达水平的变化。结果10GyX射线照射A549细胞株后14h凋亡率达到高峰。Westernblot检测显示,随着放射剂量的增加,α-dystrobrevin有下降的趋势,而ε-dystrobrevin有升高的趋势。结论人肺腺癌A549细胞株在能量为6-MV,剂量率为300cGy/分,接受10GyX线照射后14h出现明显的凋亡;并且在此时间点上随着放射剂量的增加α-dystrobrevin和ε-dystrobrevin变化趋势不同。

Objective In order to investigate the relationship between irradiation dose and cell apoptosis,the dystrobrevin was detected by Western Blot when apoptosis induced by X-ray irradiation reached the peak in lung adenocarcinoma A549 cells in vitro. Methods Being cultured for 24 hours,the human lung adenocarcinoma A549 cells were irradiated with 10Gy X-ray. The cells irradi? ated were stained by Hoechest 33258 fluorescence in 6h,14 h,23 h,40 h,while the cellmorphology changes was observed in the fluorescence microscope and the apoptosis rate was calculated. The dystrobrevin protein weredetected by Western Blot after 14 hours when the cells were irradiated by X-ray with 0Gy,6Gy and 10Gy. Results The apoptosis rate reached the peak in 14 hours after the A549 cells irradiated with 10Gy X-ray. Western blot revealed that α-dystrobrevin had a down-regulated trend and ε-dystrobrevin indicated an up-regulated trend with the doses of X-ray increase. Conclusion After human lung adenocarcinoma A549 cell was induced by 10Gy X-ray irradiation(6-MV energy,300cGy/min dose rate),apoptosis index significantly increases in 14 hours,while α-dystrobrevin and ε-dystrobrevin have different trends with the radiation doses increase.