目的观察不同加热温度及不同加热时间对人胶质瘤U251细胞凋亡的影响。方法采用恒温加热(43、44、45℃各1、2、3、4h)处理细胞,后继续培养12h。应用Annexin V-FITC/PI双染、瑞氏-姬姆萨染色、PI单染检测细胞凋亡率、凋亡细胞形态学、细胞周期生化学特征的变化情况。结果随温度升高和诱导时间延长U251细胞凋亡率增加,44℃3h凋亡率最高达40.10%(P〈0.01);光镜下可见凋亡细胞出现膜小泡,凋亡小体;PI单染有凋亡峰及细胞周期改变。结论44℃热诱导3h是促使U251细胞凋亡的最佳条件,热诱导凋亡细胞具有典型的形态、生化学特征变化。
Objective To study the effect of different temprature and time points on human glioma U251 cells apoptosis induced by hyperthermia. Methods Human U251 glioma cells were cultured in vitro and induced by byperthermia 43,44,45 ℃ for 1,2,3,4 hours respectively and then routine culture for 12 hours. The apoptotic cell morphology was observed by Wright-Giemsa staining,the percentage of the cell apoptosis and the alteration of cell cycle were detected by FCM throgh Annexin V-FTTC and PI staining. Results The percentage of the cell apoptosis increased with hightening tempreture and prolonging time and the maximum rate of apoptosis was 40.10% at 44 centi-degree for 3 hours. Typical apoptotic bodies or membranosomas appeard. Apoptotic peak and the change of cell cycle were tested by FCM. Conclusion 3 hours moderate hyperthermia at 43 centi-degree is the effective heat-dosage to induce human glioma U251 cells and the heat-induced apoptotic cells have typical morphology and biochemics character change.