联合应用反义核酸对慢性粒细胞性白血病骨髓及外周血的净化作用

The Effect of Antisense Oligodeoxynucleotides in Combination on the Purge of Bone Marrow and Peripheral Blood in Cases with Chronic Myelogenous Leukemia

目的 :比较单独及联合应用BCR_ABL反义寡核苷酸 (Antisenseoligodeoxynucleotide ,ASODN)及C_MYBASODN对慢性粒细胞性细胞白血病 (Chronicmyelogenousleukemia ,CML)细胞及正常造血细胞的抑制作用。方法 :用BCR_ABLASODN、C_MYBASODN及错配ODN ,分别对 10例CML骨髓和外周血单个核细胞 (Mononuclearcells ,MNC)及 8例正常骨髓MNC单独及联合进行处理 ,用半固体培养检测细胞集落形成率 ,并用RT_PCR法检测CML细胞BCR_ABL基因表达。结果 :3种ODN均对CML细胞集落形成及BCR_ABL基因表达有抑制作用 ,由强到弱依次为BCR_ABLASODN +C_MYBASODN组 ,二者单独应用组及错配组 (P <0 .0 5) ,三者对于正常造血细胞的集落形成也有一定抑制作用 (P <0 .0 1) ,其中联合组作用弱于C_MYBASODN组。结论 :BCR_ABLASODN及C_MYBASODN联合应用可增强它们对CML细胞生长及BCR_ABL基因表达的抑制作用 ,而对正常造血细胞的抑制作用则有所减弱。

To compare the inhibitory effects of BCR_ABL antisense oligodeoxynucleotide (AS ODN) and C_MYB AS ODN used individually and in combination on the growth of chronic myelogenous leukemia(CML)cells and normal hemotopoietic stem cells. Methods: Samples of bone marrow and peripheral blood mononuclear cells (MNC) from 10 cases with CML and 8 normal controls were treated with BCR_ABL AS ODN, C_MYB AS ODN and mismatched ODN individually and in combination respectively, then the colony_forming efficiencies of the cells were scored after being cultured in semisolid medium and the expression of BCR_ABL gene was tested by RT_PCR. Results: All of the ODNs could inhibit the colony_forming efficiency and BCR_ABL gene′s expression in CML cells. The inhibitory effect in BCR_ABL AS ODN +C_MYB AS ODN group was the strongest, that in group of each of them applied individually was weaker, and that in the group of mismatched ODN was the weakest. (P<0.05). They also had a little inhibitory effect on the colony_forming of normal hemotopoietic stem cells (P<0.01), and the effect in combination group was weaker than that in C_MYB AS ODN group (P<0.05). Conclusion: BCR_ABL AS ODN and C_MYB AS ODN used in combination could increase their inhibitory effect on the growth and BCR_ABL gene′s expression in CML cells, but decreased the inhibitory effect on normal hemotopoietic stem cells.