摘要
目的研究三氧化二砷 (As2 O3 )在体外诱导人肝癌细胞BEL 74 0 2凋亡的作用。方法用MTT、琼脂糖凝胶电泳、原位末端转移酶技术 (TUNEL)、透射电镜及流式细胞仪等方法观察细胞凋亡。结果As2 O3 在体外对BEL 74 0 2细胞生长具有显著抑制作用 ,随着作用时间的延长和As2 O3 剂量的增加 ,抑制作用随之增强。As2 O3 诱导 5 0 %以上BEL 74 0 2细胞发生凋亡的浓度为 2 μmol/L。经As2 O3 作用后的BEL 74 0 2细胞 ,在琼脂糖电泳中出现了典型的DNA凋亡条带 ,用流式细胞仪检测出了典型的凋亡峰 ,电镜下观察到了典型的凋亡细胞形态学改变 :如细胞核仁消失 ,染色质浓缩、碎裂 ,聚于核膜边缘并形成凋亡小体。结论As2 O3 在体外能有效诱导人肝癌细胞BEL 74 0 2发生凋亡。
ObjectivesTo study the apoptotic changes of human hepatocarcinoma cells (BEL 7402) induced by arsenic trioxide (As 2O 3) in vitro.MethodsThe apoptosis of BEL 7402 cells was observed by monotetrazolium (MTT) staining, agarose gel electrophoresis, TdT mediated x dUTP nick end labeling (TUNEL), transmission electron microscopy and flow cytometry method (FCM) .ResultsArsenic trioxide displayed obvious inhibition on BEL 7402 cells in vitro. Accompanying prolongation of reaction and elevated dosage of As 2O 3, the inhibition increased in its intensity. The concentration of arsenic trioxide inducing more than 50% of BEL 7402 cells to undergo apoptosis was 2 μmol/L. Typical apoptotic changes were found, such as apoptotic DNA bands appeared on agarose gel electrophoresis, apoptotic cell peak detected by FCM and morphological abnormalities seen under transmission electron microscope e.g. disappearance of cellular nucleoli, aggregation of condensed and fragmented chromatin along the margin of nucleolar membranes to form apoptotic bodies,etc.ConclusionApoptosis of human hepatocarcinoma BEL 7402 cells can be effectively induced by arsenic trioxide in vitro. [
出处
《医学临床研究》
CAS
2003年第4期252-255,共4页
Journal of Clinical Research
