DADS抑制食管胃交界部腺癌细胞OE 19增殖及诱导凋亡的机制

摘要

目的：探讨二烯丙基二硫化物(DADS)体外抑制食管胃交界部腺癌细胞株OE19增殖并诱导凋亡的作用及相关机制。方法 DADS处理 OE19,倒置显微镜下观察细胞形态变化；MTT检测 DADS对 OE19细胞的增殖抑制作用；使用流式细胞术分析不同浓度 DADS处理 OE19后的凋亡率；Real-time PCR 检测 DADS 对 OE-19细胞 Caspase-3、Caspase-9、Bcl-2、Bax以及 NF-κB mRNA表达水平的影响。结果作用24、48 h后,DADS可以抑制OE19增殖,抑制作用呈剂量依赖性。使用40、80μg/mL的 DADS 处理 OE19细胞24、48 h,凋亡细胞比例分别为14.0%、25.4%和19.0%、27.2%。Real-time PCR检测发现DADS能够上调 OE19细胞Caspase-3、Caspase-9 mRNA的表达,并且能够下调 NF-κB、Bcl-2 mRNA的表达,且呈剂量-反应关系,Bcl-2/Bax的比值明显降低。结论 DADS在体外能够抑制食管胃交界部腺癌细胞的增殖并诱导其凋亡,其凋亡机制可能是通过线粒体途径,同时 NF-κB通路以及Bcl-2家族成员均参与了该过程。%Objective To investigate the effect and mechanism of DADS in inhibiting the proliferation and inducing apoptosis of gastro-esophageal cancer cells in vitro.Methods The gastro-esophageal adenocarcinoma cells OE1 9 were treated by DADS of different concentrations in vitro.Morphologic changes were observed by the microscope and MTT assay was performed to test the growth-inhibitory effect of DADS on OE1 9 cells.Apoptosis rate of OE1 9 treated with different concentrations of DADS was measured by flow cytometry.Real-time PCR was used to detect DADS-induced effects on mRNA expressions of Caspase-3 ,Caspase-9 ,Bcl-2 ,Bax and NF-κB in OE1 9 cells.Results DADS inhibited the proliferation of OE19 cells in a dose-dependent manner.The apoptosis rate of OE19 cells was 14.0%,25.4% and 19.0% and 27.2%,respectively,when treated with 40 and 80μg/mL DADS for 24 h and 48 h.Real-time PCR assay showed that DADS could enhance mRNA expression levels of Caspase-3 and Caspase-9 and significantly decrease the mRNA expression levels of NF-κB and Bcl-2 and the ratio of Bcl-2/Bax. Conclusion DADS can significantly inhibit the proliferation and induce the apoptosis of gastro-esophageal adenocarcinoma cells via mitochondria-dependent pathways,which may be related to NF-κB and Bcl-2 families.