拓扑异构酶Ι抑制剂诱导白血病HL60/VCR耐药细胞凋亡的研究
Apoptosis of HL60/VCR Cells Induced by Topotecan
-
摘要: 目的 研究拓扑异构酶I抑制剂 (topotecan ;TPT)对白血病耐药细胞HL6 0 /VCR诱导凋亡的作用。方法 瑞氏 吉姆萨染色和吖啶橙 /溴化乙啶 (AO/EB)染色进行形态观察, 采用TUNNEL、流式细胞仪检测细胞周期和AnnexinV观察TPT对HL6 0 /VCR细胞的抑制效应和促凋亡作用。WesternBlot检测bcl 2、活化的caspase 3和P糖蛋白 (Pgp)的表达变化。 结果 经TPT处理后的HL6 0 /VCR细胞, 在光镜和AO/EB染色中均可见到典型的凋亡细胞形态学改变, 并具有时间和剂量依赖性 ;AnnexinV染色后能检测到早期凋亡细胞 (2 6 .8% ), 细胞周期显示 :G1期细胞比例增高, S期减低, 并有 2 1.8%凋亡峰 ;TUNNEL能检测到 (6 2 .2± 3.5 ) %的阳性细胞 ;伴有活化的caspase 3的表达和bcl 2的下调, 而Pgp的表达在细胞凋亡前后无变化。结论 TPT能诱导白血病耐药细胞凋亡, 该过程伴有caspase 3的活化和bcl 2的表达下调。Abstract: Objective To gain insight into the mechanisms of topotecan(TPT) inducing apoptosis of acute myelogenous leukemia multidrug-resistant HL60/VCR cells. Methods Exposed HL60/VCR cells to different concentrations of TPT, morphologic evidence for apoptosis was determined by Wright-Gimsa and Acridine Orange/ethidium bromide(AO/EB) staining. Cell cycle, Sub-G1 and Annexin V FITC staining were detected by flow cytometry. The expression of active caspase-3, bcl-2 and Pgp were detected by Western blot. Results HL...
下载:
