摘要
目的观察新型bcl-2反义寡核苷酸F951诱导人白血病细胞凋亡的作用。方法不同浓度的F951与HL60细胞共培养后,采用流式细胞术,DNA Ladder分析,电镜观察,TdT酶介导的缺口末端标记法(TUNEL)检测凋亡细胞。结果5,10,20μmol.L-1F951分别与HL60细胞共孵育48h后,线粒体凋亡细胞:未处理组、FNS对照组分别为3.00%、13.57%,F9513个剂量分别为30.95%、38.08%、52.55%;Caspase活性细胞:未处理组、FNS对照组分别为0.08%、0.14%,F9513个剂量组分别为43.68%、60.54%、80.37%。凝胶电泳分析:F951处理组可见典型的DNALadder,且随着药物浓度的提高诱导DNA Ladder的作用更加明显;TUNEL和电镜检查:未处理组与FNS对照组中仅偶见凋亡细胞,F951各剂量组凋亡细胞常见,且随着药物浓度的提高凋亡细胞增多。结论F951具有诱导HL60细胞凋亡的作用;F951诱导肿瘤细胞凋亡的作用是通过抑制bcl-2基因,启动细胞凋亡调控通道而实现的。
Aim To observe the apoptosis of human leukemic cells induced by F951, a novel bcl-2 antisense oligodeoxynucleotide. Methods HL60 cells were cultured with F951 in variant doses. Apoptotic cells were detected by flow cytometry, DNA ladder, electron microscope observation and terminal-deoxynucleotidyl transferase mediated nick end labeling （TUNEL）. Results After HL60 cells were treated for 48 hours, mitochondria apoptotic cells can be detected at the ratio of 3.00% in the untreated group, 13.57% in the FNS control group and 30.95%, 38.08%, 52.55% with 5, 10, 20 μmol·L^-1 F951 respectively; the ratio of cells with caspase activity was 0. 08% in the untreated group, 0. 14% in the FNS control group and 43.68%, 60. 54%, 80. 37% with 5, 10,20 μmol· L^-1 F951 respectively. Typical DNA ladder was seen from gel electrophoresis in F951 treated groups, and more apparent effect in the aspect of inducing DNA ladder had been observed with the improvement of F951 concentration. Detected through TUNEL and electron microscope, apoptotic cells in untreated group and FNS control group can only be found by chance, but very commonly seen in F951 treated groups and more frequently with the improvement of F951 concentration. Conclusion F951 can induce cell apoptosis in HL60 cells. Such effect is achieved through the inhibition of bcl-2 gene expression by F951 which initialize apoptosis passageway in consequence.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2007年第2期260-263,共4页
Chinese Pharmacological Bulletin
