摘要
目的探讨Ｂｃ１－２反义硫代磷酸寡脱氧核苷酸（ＡＳ－ＰＳ－ＯＤＮ，ＡＳＰＯ）是否抑制ＨＬ６０细胞Ｂｃ１－２的ｍＲＮＡ及其蛋白的表达，以及不同浓度ＡＳＰＯ产生抑制作用的程度、效应发生和持续时间的差异。方法应用免疫细胞化学染色、流式细胞仪和ＲＴ－ＰＣＲ半定量法对与Ｂｃ１－２的ＡＳＰＯ共培养后的ＨＬ６０细胞Ｂｃ１－２蛋白及ｍＲＮＡ表达进行检测。结果Ｂｃ１－２的ＡＳＰＯ能在ｍＲＮＡ水平产生抑制作用，并减少Ｂｃ１－２蛋白的合成，且在培养２４小时即出现效应，同时随着ＡＳＰＯ剂量的增加和作用时间的延长其抑制作用更显著，但７２小时后出现回升现象。结论Ｂｃ１－２的ＡＳＰＯ能特异抑制ＨＬ６０细胞Ｂｃ１－２的ｍＲＮＡ和蛋白表达，且具浓度和时间的依赖性，同时又存在作用暂时性的问题。
Objective To observe whether the expression of Bc1 2 mRNA and protein in HL60 cells could be decreased by incubation with antisense phosphorothioate oligodeoxnucleotides (ASPO) at different concentrations (5μmol/L, 10μmol/L, and 20μmol/L). Methods The expressions of Bc1 2 protein and mRNA in HL60 cells were individually determined by immunocytochemistry, flow cytometry and RT PCR. Results In the presence of ASPO, the levels of Bc1 2 mRNA and protein could be decreased. The inhibitory effect appeared in the first 24 hours of incubation and became intense as the concentration increased and the time prolonged, but it abated after 72 hours of incubation. Inhibition was not noted in the presence of sense phosphorothioate oligodeoxynucleotides (SPO). Conclusion The data suggested that ASPO could inhibit the expression of Bc1 2 mRNA and protein in HL60 cells and the inhibitory effect depended on the specific sequences, the concentration and the time of incubation, but it could not last very long.
出处
《中华医学遗传学杂志》
EI
CAS
CSCD
北大核心
1999年第1期12-15,共4页
Chinese Journal of Medical Genetics
基金
福建省科委优先发展基金
关键词
ASPO
BCL-2
表达
HL60细胞系
基因治疗
Antisense phosphorothioate oligodeoxynucleotides Bcl 2 Expression HL60 cell line
