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目的：研究 ｂｃｒ－ａｂｌ融合基因硫代磷酸反义寡核苷酸（Ａｓｐｏ）对 Ｋ５６２细胞生长特性的影响，探讨其在慢性粒细胞白血病（ＣＭＬ）基因治疗中的意义。方法：倒置显微镜下细胞活体观察；Ａｓｐｏ与细胞共同培养２４ｈ、４８ｈ、７２ｈ、９６ｈ及 １２０ｈ，用０．４％台盼蓝染色计各处理组死活细胞数，甲基纤维素半固体培养法观察其克隆形成率的变化，流式细胞仪测定各处理组细胞Ｐ２１０蛋白表达变化。结果：Ａｓｐｏ处理后细胞仍呈克隆状生长，当 Ａｓｐｏ浓度达５μｍｏｌ／Ｌ时，即可产生增殖抑制作用，呈一定的量效关系，其最大抑制效应时间为作用１２０ｈ，当Ａｓｐｏ达１０μｍｏｌ／Ｌ时，在一定细胞浓度范围内（１×１０４／ｍＬ－ ５× １０５／ｍＬ），均有显著抑制作用，当Ａｓｐｏ浓度达５μｍｏｌ／Ｌ以上时，Ｋ５６２细胞 Ｐ２１０蛋白表达明显下降甚至不表达。 ｂ２ａ２型 Ａｓｐｏ对表达 ｂ３ａ２型 ｍＲＮＡ的 Ｋ５６２细胞也表现出交叉抑制作用。结论：ｂｃｒ－ａｂｌ融合基因反义寡核苷酸对Ｋ５６２细胞具有特异性增殖抑制作用，在慢性粒细胞白血病基因治疗中值得深人研究。
OBJECTIVE: To study the effect of bcr-abl fusion gene phosphorothioate antisense oligonucleotide (Aspo) on the growth of K562 cells and to explore its significance in the gene therapy of chronic myelogenous leukemia (CML). Methods: The cells were observed under inverted microscope. Aspo cells were incubated with cells for 24h, 48h, 72h, 96h and 120h. The numbers of viable cells in each treated group were measured by 0.4% trypan blue staining, and the effect of methylcellulose semi-solid culture The clonogenic rate of the cells was determined by flow cytometry. Results: Aspo treated cells still showed clonal growth. When Aspo concentration reached 5μmol / L, the proliferation could be inhibited, with a dose-effect relationship. The maximum inhibitory effect time was 120h. When Aspo reached 10μmol / L (1 × 104 / mL-5 × 105 / mL). When Aspo concentration was above 5μmol / L, the expression of P210 in K562 cells was significantly decreased or even not expressed. Aspartate b2a2 also showed cross-inhibitory effects on K562 cells expressing b3a2 mRNA. CONCLUSION: The bcr-abl fusion gene antisense oligonucleotide has a specific inhibitory effect on K562 cells, which deserves further study in gene therapy of chronic myeloid leukemia.