去甲斑蝥素诱导人食管癌Eca-109细胞凋亡及其作用机制  

Effect of norcantharidin on apoptosis of human esophageal cancer cell Eca-109 and its possible mechanism

作　　者：许欣[1] 彭林涛[2] 

机构地区：[1]邢台市人民医院病理科,邢台054031 [2]邢台市人民医院外科,邢台054031

出　　处：《肿瘤》2010年第8期666-670,共5页Tumor

摘　　要：目的:探讨去甲斑蝥素诱导人食管癌Eca-109细胞的凋亡及其可能的作用机制。方法:去甲斑蝥素作用食管癌Eca-109细胞后,应用MTT法检测其对细胞的生长抑制作用,透射电子显微镜下观察细胞超微结构的变化,琼脂糖凝胶电泳观察其对细胞凋亡的诱导作用,RT-PCR法检测caspase8和caspase3mRNA的表达,Western印迹法检测Fas、细胞型含死亡域的Fas结合蛋白样白介素-1β转换酶抑制蛋白(cellular FADD-like interleukin-1βconverting enzyme inhibitory protein,c-FLIP)、caspase8和caspase3蛋白的表达。结果:去甲斑蝥素对人食管癌Eca-109细胞具有生长抑制作用,并呈时效和量效依赖关系。电子显微镜下观察发现,食管癌Eca-109细胞趋于凋亡。DNA琼脂糖凝胶电泳可见典型的DNA梯状条带。RT-PCR检测显示,caspase8和caspase3mRNA的表达水平明显上升(P<0.01)。Western印迹法检测结果显示,与阴性对照组比较,去甲斑蝥素作用后,食管癌Eca-109细胞中Fas、caspase3和caspase8蛋白的表达明显上升(P<0.05),c-FLIP蛋白的表达水平明显下降(P<0.05)。结论:去甲斑蝥素能够抑制食管癌Eca-109细胞的生长并诱导其凋亡,其机制可能是通过上调Fas、caspase8、caspase3的表达和下调c-FLIP的表达来实现的。Objective ： To investigate the effect of norcantharidin in inducing apoptosis of human esophageal cancer Eca-109 cells and elucidate its possible molecular mechanism. Methods： Cell growth was detected by MTT assay after norcantharidin treatment. The ultrastructural alterations were observed under transmission electron microscope （TEM）. The cell apoptosis was determined by agorase gel electrophoresis. The mRNA expression levels of caspase 8 and caspase 3 were detected by RT-PCR and the protein expression of Fas, cellular FADD-like interleukin-1β converting enzyme inhibitory protein（c-FLIP） , caspase 8, and caspase 3 were determined by Western blotting. Results：Norcantharidin inhibited growth of Eca-109 cells in a dose-and time-dependent manner. Distinct morphological changes of apoptosis were found in norcantharidin-treated cells under TEM. A characteristic DNA ＂ladder＂ on the agrose gels was observed. RT-PCR showed that the mRNA expression of caspase 8 and caspase 3 markedly increased （P 〈 0.01 ）. Western blotting demonstrated that the protein expression of Fas, caspase 8 and caspase 3 greatly increased after norcantharidin treatment compared with negative control group （ P 〈 0.05 ）, but the expression of c-FLIP protein decreased （ P 〈 0.05 ）. Conclusion ： Norcantharidin inhibits the apoptosis and induces apoptosis of esophageal cancer Eta-109 cells through up-regulation of Fas, caspase 8, and caspase 3 and down-regulation of c-FLIP expression.

关 键 词：食管肿瘤 细胞凋亡 基因表达调控 去甲斑蝥素 细胞 ECA-109 

分 类 号：R735.1[医药卫生—肿瘤]