^(32)P胶体体外诱导颅咽管瘤细胞凋亡

^(32)P colloid induced apoptosis of craniopharyngioma cells in vitro

目的:探讨32P胶体对颅咽管瘤(craniopharyngioma,CP)体外培养细胞诱导凋亡作用及剂量效应和时间效应关系。方法:通过原代细胞培养获得CP有限传代细胞系,经不同浓度32P胶体处理不同时间后,应用MTT比色法绘制细胞存活率曲线。流式细胞仪定量检测细胞凋亡率。Hoechst 33342荧光染色检测凋亡细胞形态。TUNEL荧光染色检测DNA特征改变。透射电子显微镜(TEM)检测细胞超微结构。结果:Hoechst 33342荧光染色、TUNEL荧光染色、TEM均证实32P胶体确能引起CP细胞产生凋亡。随着32P胶体处理浓度(0~14.80 MBq/mL)及时间(1~14 d)的增加,CP细胞存活率减少、凋亡率增高。结论:32P胶体能明显抑制CP体外培养细胞生长并诱导凋亡,剂量越大及处理时间越长其杀伤作用越强。

Objective：This study aimed to investigate the possible mechanism of 32P colloid induced apoptosis of craniopharyngi-oma （CP） cells in vitro and the relationship between dose effect and time effect. Methods：This study established a primary cell culture of CP limited subculture cell line. Methyl thiazolyl tetrazolium （MTT） assay was performed to plot the cell survival curve after the CP cells were treated with 32P colloid at different concentrations and time. Apoptotic rate was detected by flow cytometry（FCM）. Apoptosis related DNA was investigated by TUNEL fluorescent staining. The morphological characteristics of apoptotic cells were determined by Hoechst33342 fluorescence staining. The ultrastructure of apoptotic cells was investigated by transmission electron microscopy （TEM）. Results：Hoechst33342 fluorescence staining, TUNEL fluorescence staining, and TEM revealed that 32P colloid induced the apoptosis of CP cells. 32P colloid reduced the survival rate and increased the apoptotic rate of CP cells as concentration （0 MBq/mL to 14.80 MBq/mL） and time （1 d to 14 d） were increased. Conclusion： 32P colloid could effectively inhibit the growth of CP cells and induce apoptosis in vitro. High concentrations and prolonged time could induce a remarkable effect.