摘要
目的:观察刺果紫玉盘素J(Calamistrin J,Cal-J)诱导人结肠癌Lovo细胞凋亡的作用,并探讨其诱导细胞凋亡的可能机制。方法:Hoechst33258染色后在荧光显微镜下观察凋亡细胞的形态变化,流式细胞仪(FCM)分析细胞DNA含量变化及计算凋亡率。荧光酶标仪检测DCFH-DA荧光探针标记的活性氧(ROS),以DiOC6(3)标记、流式细胞仪测定细胞线粒体膜电位(Δψm)的改变。结果:Cal-J作用48 h后,荧光显微镜下可见细胞核固缩,染色质凝聚等典型凋亡形态学特征。FCM检测可见Lovo细胞凋亡率明显增加,且具有时间、剂量的双重依赖性。Cal-J处理Lovo细胞12 h后,可剂量依赖性地增高胞内ROS水平,Cal-J 100μg.mL-1作用于Lovo细胞,可时间依赖性降低Δψm。结论:Cal-J具有诱导Lovo细胞凋亡的作用,该作用呈一定的剂量和时间依赖性,其机制与提高细胞内ROS水平和降低线粒体膜电位有关。
Aim：To observe the apoptosis induced by Calamistrin J on human colon carcinoma Lovo cells, and to explore its mechanism. Methods： Observe the morphological changes of the apoptosis cells by fluorescence microscope with Hoechst33258 staining, measure the apoptosis rate of Lovo cells by flow cytometry（FCM）. ROS levels in cells labelled by DCFH-DA were assayed by spectrofluorometer and the change of △ψm was analyzed by FCM with DiOC6 （ 3 ） staining. Results： Lovo cells treated by Calamistrin J could be observed to have typically morphological changes through fluorescence microscope, including cell shrinkage with a condensed cytoplasm , chromatin condensation, etc. It was measured by FCM that Calamistrin J induced apoptosis in a time-and dose-independent manner. The cellular ROS level of Lovo cells showed a significant increase after exposed to Calamistrin J for 12 h in a dose- independent manner. It was observed that△ψm began to decrease after exposed to Calamistrin J 100 μg · mL^-1 , the loss of △ψm become more remarkable as time goes on. Conclusion： Calamistrin J significantly induced apoptosis of Lovo cells, which may be due to its increase of the cellular ROS level and the decrease of △ψm.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2007年第2期142-146,共5页
Journal of Jinan University(Natural Science & Medicine Edition)
关键词
刺果紫玉盘素J
结肠癌
凋亡
活性氧
线粒体膜电位
Calamistrin J
colon carcinoma
apoptosis
reactive oxygen species
mitochondrialmembrane potential
