摘要
目的 探讨Survivin反义核酸对紫杉醇诱导白血病细胞系HL 6 0细胞凋亡的影响。方法 将已构建成功的Survivin反义核酸真核表达载体pcDNA3 SVVas电转染HL 6 0细胞 ,并筛选转染成功的阳性克隆 ;锥虫蓝拒染法观察Survivin反义核酸与紫杉醇联合应用对HL 6 0细胞增殖的影响 ;细胞计数和MTT实验测定转染细胞对紫杉醇敏感性 ;琼脂糖凝胶电泳分析细胞凋亡DNA断裂情况 ;核染色检测凋亡细胞的细胞核变化。结果 转染Survivin反义核酸的HL 6 0SVVas细胞增殖明显受抑 ,与HL 6 0neo细胞、HL 6 0细胞进行比较 ,差异具有显著性 (P <0 .0 5 ) ;MTT实验显示 ,紫杉醇对HL 6 0SVVas、HL 6 0neo、HL 6 0细胞的IC50 值分别为 (14 .4± 1.87)ng ml,(31.9± 6 .38)ng ml和 (32 .0± 6 5 2 )ng ml。统计学分析证明差异具有显著性 (P <0 .0 1) ;经琼脂糖凝胶电泳 ,HL 6 0SVVas细胞可见到DNA梯形条带 ,而HL 6 0neo、HL 6 0细胞未见到 ;与HL 6 0neo、HL 6 0细胞相比 ,HL 6 0SVVas的细胞核呈致密浓染。结论 Survivin反义核酸可促进紫杉醇诱导HL 6 0细胞凋亡。
Objective To explore the effects of survivin antisense RNA on taxol induced apoptosis in leukemia cell line HL 60. Methods A survivin antisense eukaryotic vector pcDNA3 SVVas was transferred into HL 60 cells by electroporation.The live fraction was determined by trypan blue dye exclusion assay.Cell counting and MTT assay were performed to evaluate the sensibility of the transfected cells to taxol. Apoptosis was detected by DNA gel electrophoresis and nuclear staining.Results Two positive cell clones, HL 60 SVVas and HL 60 neo were obtained. Compared to HL 60 and HL 60 neo cells, HL 60 SVVas cells growth was significantly reduced (P<0 05).By MTT assay, the IC 50 of taxol to HL 60 SVVas, HL 60 neo and HL 60 cells were (14.4±1.87)ng/ml, (31.9±6.38)ng/ml and (32.0±3.52)ng/ml, respectively,the difference was significant by statistic analysis(P<0 01). Agarose gel electrophoresis of genomic DNA from HL 60 SVVas showed typical DNA ladder,but DNA from HL 60 neo and HL 60 did not. Nuclei become condense in HL 60 SVVas cells.Conclusion Survivin antisense RNA could enhance taxol induced apoptosis in leukemia cell line HL 60.This may lay an experimental foundation for further research of gene therapy in leukemia.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2003年第7期351-354,共4页
Chinese Journal of Hematology
基金
国家自然科学基金资助项目 ( 3 9970 693 )
