^(32)P-磷酸铬体外诱导人非小细胞肺癌A549细胞凋亡

Study on apoptosis of human non-small cell pulmonary carcinoma A549 cells induced by ^(32)P-chromium phosphates in vitro

目的 观察32 P 磷酸铬 ( 32 P胶体 )体外诱导人非小细胞肺癌 (NSCLC)A5 4 9细胞周期变化和发生细胞凋亡的现象 ,建立剂量 效应和时间 效应关系。方法 将32 P胶体加入A5 4 9细胞培养体系进行内照射 ,初始放射性浓度分别为 0 ,93,180 ,2 78,370和 4 6 3MBq L ,采用Giemsa染色、透射电镜、末端原位标记 (TUNEL)等方法进行凋亡细胞的形态学、超微病理、生化特征检测 ,应用流式细胞术定量检测细胞周期变化和细胞凋亡率。结果 A5 4 9细胞受照射后S期 +G2 M期细胞比率在 96h以内呈上升趋势 ,之后逐渐下降。A5 4 9细胞受照射后 72h左右出现兴奋性表现 ,96h起各受照组均检出凋亡 ,各剂量组在照射后 12 0h达到凋亡高峰。结论 在较低剂量范围内 ,32 P胶体内照射可以体外诱导人NSCLCA5 4 9发生 72h以后的迟发相凋亡 。

Objective To observe the phasic change and apoptosis of A549 non-small cell lung cancer cells induced by 32 P chromium phosphate in vitro ,and establish the dose-response and time-response relationship. Methods Internal irradiation was conducted by adding 32 P colloid into A549 cell culture system.The initial radioactivities were 0,93,180,278,370,463 MBq/L,respectively.Giemsa stain,transmission electron microscopy and TUNEL were used in assessing morphologic,ultra structural pathologic and biochemical characteristics,and the phasic changes and apoptotic rates of cells were studied by flow cytometry. Results After irradiation of A549 cells,the cell ratio of S+G 2 M phase tended to increase within 96 h,then decreased gradually.At 72 h after irradiation the A549 cells showed excited manifestation,and in each irradiation group apoptosis began from 96 h of irradiation,and attained the peak at 120 h. Conclusion In the lower dosagerange, 32 P internal irradiation may induce human NSCLC A549 cells to present delayed onset of apoptosis,and the rate of cell apoptosis is positively correlated to the initial radioactivity concentration.