反义BCR/ABL寡核苷酸体外抑制K562细胞的研究

In Vitro Inhibition of K562 Cell Lines by BCR ABL Antisense Oligonucleotide

目的：探讨反义ＢＣＲ和ＡＢＬ融合基因（ＢＣＲ／ＡＢＬ融合基因）寡核苷酸体外抑制Ｋ５６２细胞的作用及机制。方法：采用Ｋ５６２细胞培养，观察反义ＢＣＲ／ＡＢＬ寡核苷酸体外对Ｋ５６２细胞数、台盼蓝拒染率及克隆形成等的影响。结果：经反义ＢＣＲ／ＡＢＬ寡核苷酸ｂ３ａ２（ＡＳｂ３ａ２）处理后培养８ｄ的Ｋ５６２克隆抑制率达６０６９％，液体培养中细胞数从２４ｈ开始较对照组减少，细胞存活率从８ｈ开始即较对照组明显下降，ＢＣＲ／ＡＢＬ寡核苷酸ｂ２ａ２（ＡＳｂ２ａ２）及无义寡核苷酸对Ｋ５６２细胞数和存活率无明显影响；３种寡核苷酸对ＨＬ６０细胞数和存活率也无影响。结论：ＡＳｂ３ａ２对Ｋ５６２细胞有序列特异性的抑制作用。ＡＳｂ３ａ２的这种抑制作用可能在于它诱导Ｋ５６２细胞的凋亡。

Objective: To explore the inhibition of K562 cell lines by bcr abl antisense oligonucleotide in vitro. Methods: K562 cell line was used to observe the influences on cell proliferation, the ability to exclude trypan blue and the clonal formation in vitro treatment with BCR/ABL antisense oligonucleotide. Results: The clonal formation of K562 cell line after treatment with BCR/ABL antisense oligonucleotide b3a2 (AS b3a2) was inhibited obviously. In liquid culture, the quantity of cell proliferation decreased from 24 hour after treatment, while the ability to exclude trypan blue significantly decreased from 8 hours after treatment. No influences on the number and activities of K562 cells were found when the cells were treated with BCR/ABL antisense oligonucleotide b2a2 (AS b2a2) or non sense oligonucleotide. Meanwhile, these three oligonucleotides could not influence the number and activities of HL 60 cell lines. Conclusions: AS b3a2 could sequence specifically inhibit activities and proliferation of K562 cells.