二硫苏糖醇诱导Eca109细胞凋亡及P38磷酸化检测

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／LDTT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。

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

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.