高温对HeLa细胞的杀伤作用

The effect of hyperthermia on HeLa cells

目的 探求不同高温对肿瘤细胞的杀伤作用。方法 将人子宫颈癌细胞系(HeLa细胞)分为37℃,40℃,45℃,50℃,55℃和60℃六个温度组,每组又分为 15 min,30 min,1 h,2 h和4 h五个时间亚组。经相应的温度和时间处理后,以四甲基噻唑兰(MTT)比色法检测细胞活性,吖啶橙荧光染色法观察细胞的形态学改变。结果 ①细胞活性检测:不同温度对HeLa细胞的杀伤作用不一样。40℃时细胞平均死亡率为15％,45℃时为54％,50℃时加温 15 min细胞全部死亡。在一定的温度下,伴随作用时间的延长,细胞死亡率也随之增加。如 45℃时,15 min的细胞死亡率为 32％、30 min的死亡率为 40％、1 h的死亡率为 56％、2 h的死亡率为68％、4 h的死亡率为74％。②形态学观察:40℃组,作用 1 h以下者无明显的形态学改变,2 h,4 h时可见部分细胞呈现凋亡。45℃组,15 min,30 min时均见大量细胞呈现凋亡。1 h,2 h,4 h则以坏死为主。50℃以上组,大部分细胞坏死。结论 随着温度的增高其对HeLa细胞的杀伤作用逐渐增强。40℃以下没有明显的杀伤作用,40℃～45℃之间,细胞的死亡形式以凋亡为主,45℃以上主要为坏死。

Objective To study the effect of hyperthermia on HeLa cells. Methods The HeLa cells were divided into six groups (37℃,40℃,45℃, 50℃, 55℃ and 60℃) and each group was divided into five subgroups (15min, 30min, 1h,2h and 4h ) and treated with the preset temperature for different period of time accordingly. Then the activity of HeLa cells was tested by MTT method and the cells were stained by acridine orange (AO) and ob- served by microscopy. Results The killing effect on HeLa cells was difference for different temperatures. The aver- age death rates of HeLa cells, were 15% in 40℃, 54% in 45℃, 100% in 50℃. And for a certain preset tempera- ture, the longer the action time, the higher the death rate of the HeLa cell. When treated with the temperature of 45℃, the death rates of HeLa cells, were 32% at 15min. 40% at 30min, 56% at 1h, 68% at 2h, 74% at 4h. At 40℃, the cells showed no morphological change for an action time of 15min, 30min and 1h; But for 2h and 4h, some cells showed apoptosis. At 45℃, many cells showed apoptosis at 15min and 30min. At or above 50℃, most cells showed necrosis. Conclusion The killing effect of hyperthermia on HeLa cells was positively correlated with tem- perature. Below 40℃, there was no killing effeet. Between 40℃～45℃, the main form of cell death is apoptosis. Above 50℃, the main form of cell death is necrosis.