摘要
目的观察醋酸棉酚对人Burkitt淋巴瘤细胞株Raji细胞增殖和凋亡的影响。方法采用台盼蓝染色和噻唑蓝(MTT)法观察醋酸棉酚对Raji细胞生长存活的影响,瑞氏染色法观察药物处理后细胞形态学变化,琼脂糖凝胶电泳和Annexin V-FITC标记流式细胞仪检测细胞凋亡,流式细胞仪分析细胞周期、检测细胞凋亡率及Bcl-2蛋白表达水平变化,比色法测定Raji细胞Caspase-3样蛋白酶活性。结果5μmol/L以上浓度醋酸棉酚能抑制Raji细胞生长增殖并诱导其凋亡,效应与药物浓度及给药时间呈正相关。醋酸棉酚作用后,Raji细胞被阻滞于G0/G1期。比色法显示Caspase-3样蛋白酶活化。流式细胞仪检测到Bcl-2蛋白表达下调,且与Caspase-3活化在时间上存在同步趋势。结论醋酸棉酚能抑制Raji细胞增殖并诱导凋亡,其原因可能与调节细胞周期及下调Bcl-2蛋白表达有关。
Objective To investigate the effects of gossypol acetate on proliferation and apoptosis in Raji lymphoblastoid cells and explore the possible mechanism. Methods Trypan blue staining and ethyhhiazolyldiphenyl-tetrazolium bromide （MTF） assay were performed to measure the effect of gossypol acetate on the growth of Raji cells. The morphologic changes were observed with Wright＇s staining assay. Apoptosis was identified by agarose-gel electrophoresis and annexin V-FITC marked flow cytometry （FCM） analysis. The distribution of cell cycle, apoptosis rate, and Bcl-2 protein expression were analyzed by FCM. Caspase-3 activity was detected by colorimetric assay. Results Gossypol acetate inhibited proliferation and induced apoptosis of Raji cells at concentration higher than 5 p, mol/L. The effects were both dose- and time- dependent. Cycle analysis indicated the alteration of cell cycle and G0/G1 arrest. The activation of Caspase-3 was observed by colorimetric assay. The results of flow cytometry showed that the down-regulation of Bcl-2 protein expression and the activation of Caspase-3 seemed to occur simultaneously. Conclusions Gossypol acetate can inhibit the growth of Raji cells and induce their apoptosis. The mechanism may be related to the alteration of cell cycle and the down- regulation of Bcl-2 protein expression.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2009年第5期527-532,共6页
Acta Academiae Medicinae Sinicae
