摘要
目的：研究ＨＬ－６０细胞中ｂｃｌ－２基因的表达并探索ｂｃｌ－２基因ｍＲＮＡ特异的反义寡聚脱氧核苷酸（反义ＯＤＮ）对ＨＬ－６０细胞中ｂｃｌ－２基因表达的影响．方法：采用细胞原位杂交和免疫细胞化学染色技术检测ＨＬ－６０细胞中ｂｃｌ－２基因的表达，人工合成ｂｃｌ－２ｍＲＮＡ特异性反义ＯＤＮ片段与ＨＬ－６０细胞共培养，在作用一定时间后分别测定细胞中ｂｃｌ－２ｍＲＮＡ和Ｂｃｌ－２蛋白表达的变化．结果：ｂｃｌ－２基因在ＨＬ－６０细胞中既有转录水平的高表达，又有翻译水平的高表达，人工合成的反义ＯＤＮ片段在一定浓度范围内，作用一段时间后细胞内ｂｃｌ－２ｍＲＮＡ表达无显著改变，而蛋白表达却呈现了显著下降，与浓度和作用时间呈正相关．结论：人工合成的反义ＯＤＮ片段可以抑制ＨＬ－６０细胞中ｂｃｌ－２的转录作用．在进行白血病基因治疗的实验研究中，ｂｃｌ－２基因可以作为基因治疗的一个有用的靶基因．
Objective:It is aimed at investigating the expression of bc-2 oncogene in HL-60 cells,then the ef-fect of an antisense oligodeoxynucleotid specific to bcl-2 mRNA on the expression of this gene in HL-60 cells.Metbods:Primarilly,in situ hybridization method and immunocytochemical staining assay were used to detect the expressional level of bcl-2 mRNA and protein separately.secondly,according to known sequence of bcl-2 mRNA,we artificially synthesized bcl-2 specific antisense oligodeoxynucleotids which were added to in uitro HL-60 culture system at different concentration in order that these antisense oligomers were introduced into cytoplasm of HL-60 cells.At given times,both the cellular bcl-2 mRNA and protein expression were detect-ed by the methods as same as primarily.Results:It is found that HL-60 cells possessed higher expression of bcl-2 gene both at mRNA and protein level.Expression of bcl-2 mRNA in HL-60 was observed no changing remarkably after adding the antisense oligomers,on the other hand,expression of bcl-2 protein was noticed declining significantly to some extent by the dose-dependent manner and time-dependent manner.Couclu-slons:bcl-2 mRNA specific antisense oligodeoxynucleotid could inhibit the expression of Bcl-2 protein in HL-60 cells.In future gene therapy research for leukemia,the oncogene bcl-2 may be choiced as a useful target.
出处
《第四军医大学学报》
1996年第2期132-134,共3页
Journal of the Fourth Military Medical University
