二硫苏糖醇诱导Eca109细胞凋亡及P38磷酸化检测

Detection of phosphorylated P38 MAP kinase in human esophageal carcinoma Eca109 apoptotic cells induced by DTT

目的:检测二硫苏糖醇(DTT)诱导Eca109细胞凋亡及磷酸化P38(PP38)水平。方法:取对数生长期Eca109细胞分为3组:2mmol/L DTT处理组、PP38抑制剂SB203580孵育细胞2h后再加DTT处理组及对照组。应用流式细胞仪检测各组细胞凋亡率;采用免疫组织化学技术检测P38的磷酸化水平。结果:DTT组、SB203580+DTT组及对照组的细胞凋亡率分别为16.8%、7.8%和3.1%。DTT组和DTT+SB203580组细胞凋亡率均高于对照组(P<0.01);与DTT组相比,SB203580+DTT组凋亡率下降(P<0.01)。DTT组PP38水平高于对照组(P<0.01)。结论:DTT可诱导人食管癌Eca109细胞凋亡,P38磷酸化起促进凋亡的作用。

Aim ： To study the role of phosphorylated p38 （ PP38 ） MAP kinase in DTT induced apoptosis of Eca109 cell line. Methods： Eca109 cells were treated with 2 mmoL/L of DTT,or 2 mmoL/L of DTT after 2 h incubation with 10 mg/L of PP38 inhibitor SB203580 for 24 h. The cells treated with saline were used as control. PP38 MAP kinase was detected using immunohistochemistry and apoptosis rate was examined using flow cytometry （ FCM ）. Results ：DTT and SB203580 plus DTT induced Eca109 cells apoptosis at the rate of 16.8% and 7.8% ,respectively,which were significantly higher than that （3.1% ） in control group（P 〈 0.01 ）, and SB203580 plus DTT could obviously decrease the apoptosis rate compared with DTT（ P 〈 0.01 ） ;the phosphorylation level of P38 in control group was significantly lower than that in DTT group （P 〈 0.01 ）. Conclusion ： The PP38 MAP kinase plays an essential role in promoting Eca109 cell apoptosis induced by DTT.