硫代反义bcl-2脱氧寡核苷酸对U937细胞凋亡的影响

Effects of Phosphorothioate Antisense bcl-2 Oligodeoxynucleotides on Human Monocytic U937 Cells Apoptosis

为研究硫代反义ｂｃｌ－２ 脱氧寡核苷酸（ＡＮ－ Ｓ－ ＯＤＮｓ） 对单核白血病细胞系Ｕ９３７ 细胞株凋亡的影响，采用Ｎｏｒｔｈｅｒｎ ｂｌｏｔ、ＤＮＡ 电泳、电镜和流式细胞计数技术检测了单核白血病细胞系Ｕ９３７ 细胞株ｂｃｌ－ ２ 基因的表达和ＡＮ－ Ｓ－ ＯＤＮｓ 对Ｕ９３７ 细胞凋亡的影响。结果发现Ｕ９３７ 细胞有大量ｂｃｌ－ ２ 基因表达；适当浓度（３０μｍｏｌ／Ｌ）ＡＮ－ Ｓ－ ＯＤＮｓ 使Ｕ９３７ 细胞出现凋亡ＤＮＡ 电泳梯状条带，电镜下可见部分细胞核浆比例缩小，核膜周边有核小体，流式细胞计数显示ｂｃｌ－ ２ 蛋白表达降低，细胞增殖能力降低。说明适当浓度的ＡＮ－ Ｓ－ ＯＤＮｓ 可以通过大大降低ｂｃｌ－２ 的表达而促进Ｕ９３７ 细胞凋亡。

The expression of bcl-2 oncogene in U937 cells,and the effects of phosphorothioate antisense oligodeoxynucleotides (AN-S-ODNs) on expression of this gene in U937 cells were investigated.Northern blot,DNA gel electrophoresis,electronmicroscope and flow cytometry technique were used to detect bcl-2 mRNA and its protein respectively,and also observed the effects of synthetic bcl-2 AN-S-ODNs on U937 cells apoptosis.It was found that U937 cells possessed high expression of bcl-2 gene both at mRNA and protein level.Proper concentration of AN-S-ODNs (30 μmol/L) could down-regulate significantly its protein level .Apoptotic cell were seen by electronmicroscope,and DNA ladder appeared during electrophoresis analysis.The results suggest that the AN-S-ODNs at higher concentration could inhibit the expression of bcl-2 protein in U937 cells,the decrease strongly of bcl-2 gene expression might promote the U937 cells apoptosis.