人肺癌细胞H1299放射敏感性与细胞周期研究

目的通过对人肺癌细胞H1299照射前、后细胞周期分布及其周期相关基因状态等研究,初步探讨其放射敏感性与照射后细胞周期阻滞的关系。方法利用克隆形成法比较H1299与H1299细胞照射后细胞存活份数及流式细胞术研究其细胞周期分布的差异;用功能基因芯片技术对周期相关的421基因进行分析研究;流式细胞术测定细胞周期及凋亡;苔盼蓝染色研究细胞死亡方式;用克隆形成率检测细胞的存活曲线。结果H1299细胞对γ射线高度敏感,照射后G1期阻滞消失,但仍存在G2期阻滞;基因芯片分析,其中周期相关基因有13个基因表达改变超过对照组2倍以上;苔盼蓝染色检测到的细胞死亡比例与对照组比较差异无显著性。结论结果表明射线对细胞的杀伤绝大部分不是由于照射后细胞凋亡所致,而是由于基因表达改变使其细胞周期阻滞,失去增殖能力引起增殖死亡的结果。

Objective To explore the relationship of radiosensitivity and postirradiational cell cycle blockage of human lung carcinoma cell H1299 . Methods Clone formation was used to compare the different counts of the survival cells between H1299 cells pre - irradiation and post - irradiation , and the difference of cell cycle distribution between them was analyzed by flow cytometry ; the cell cycle related gene - 421 gene was analyzed with gone array; cell cycle and apoptosis were determined by flow cytometry ; manners of cell death were studied with trypan blue staining ; rate of clone formation was used to test the cells＇ survival curve . Results H1299 was highly sensitive to 7 rays , the G1 phase blockage disappear after irradiation yet G2 phase blockage still existed ; the result of gone array showed there was more than twice in change of gene expression in 13 cell cycle related genes in the experiment group than control ; there was no significant difference in cell death ratio tested by trypan blue staining between the experiment group and control group . Conclusion The cell reduction after irradiation is not mostly due to the apoptosis after irradiation but due to the change of gone expression ,which block the cell cycle therefore depriving the cells＇ of their proliferation activity.