热疗通过提高活性caspase-3的表达促进食管癌EC109细胞凋亡

Hyperthermia induces apoptosis of EC109 cells by increasing the expression of activited-caspase-3

目的:探讨不同温度热疗诱导食管癌EC109细胞的生长抑制、细胞凋亡及其与天冬氨酸特异性半胱氨酸蛋白水解酶-3(caspase-3)蛋白表达变化的关系。方法:采用PCR仪精确控制温度,对食管癌EC109细胞行37℃(对照),43℃,45℃,47℃热疗处理,四甲基偶氮唑蓝(Tetrazolium bromide,MTT)法检测细胞生长抑制率,碘化丙啶(Propidium iodide,PI)/磷脂结合蛋白V(Annexin V)双染流式细胞仪测定细胞凋亡率,蛋白印迹(Western bolt)法检测caspase-3蛋白表达。结果:食管癌细胞在37℃,43℃,45℃处理下,细胞生长抑制率随温度升高而增多,并有显著性差异,43℃细胞凋亡率达到最高,三者差异显著。45℃和47℃生长抑制及凋亡率均无明显差异。总-caspase-3(pro-caspase-3)表达随温度升高而增多,而活性csapase-3(actived-caspase-3)在43℃表达最多。结论:热疗通过提高活性caspase-3表达诱导食管癌EC109细胞凋亡,在43℃凋亡率达到最高。

Objective：To explore the cell growth inhibition and apoptosis of esophageal cancer cells EC109 induced by hyperthermia in different temperature and its relation with the expression of caspase-3.Methods：Using PCR instrument to control the temperature precisely,cells treated at 37℃ as control,EC109 cells were treated at 43℃,45℃,47℃.The cell growth,apoptosis rate was detected by MTT assay and flow cytometry.The expression of caspase-3 was measured by Western Bolt.Results：The cell growth inhibition rates of EC109 cells were increased gradually at 37℃,43℃ and 45℃.The apoptosis rate at 43℃ was significantly higher than other groups.The cell growth inhibition rates and apoptosis rates had no significant difference between 45℃ and 43℃.Pro-Caspase-3 expression increased with increasing temperature,while actived-caspase-3 was expressed highest at 43℃.Conclusion：Hyperthermia induces apoptosis of EC109 cells by increasing the expression of activited-caspase-3 and the apoptosis rate reaches a peak at 43℃.