NORMAL AND LEUKEMIC HEMATOPOIETIC-CELLS MANIFEST DIFFERENTIAL SENSITIVITY TO INHIBITORY EFFECTS OF C-MYB ANTISENSE OLIGODEOXYNUCLEOTIDES - AN INVITRO STUDY RELEVANT TO BONE-MARROW PURGING
NORMAL AND LEUKEMIC HEMATOPOIETIC-CELLS MANIFEST DIFFERENTIAL SENSITIVITY TO INHIBITORY EFFECTS OF C-MYB ANTISENSE OLIGODEOXYNUCLEOTIDES - AN INVITRO STUDY RELEVANT TO BONE-MARROW PURGING
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DOI：
10.1073/pnas.88.6.2351
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发表时间：
1991-03-01
影响因子：
11.1
通讯作者：
GEWIRTZ, AM
中科院分区：
文献类型：
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作者：
CALABRETTA, B;SIMS, RB;GEWIRTZ, AM
c-myb原癌基因优先在造血细胞中表达，其编码的蛋白Myb是造血细胞增殖所需的。 为了分析正常和白血病人造血祖细胞的相对Myb依赖性，将正常骨髓细胞、几种类型的白血病母细胞以及正常和白血病细胞的1：1混合物在存在c-myb正义或反义寡脱氧核苷酸的情况下培养;然后评估细胞活力和克隆效率。 c-myb正义寡聚体对正常细胞和白血病细胞的影响可忽略不计。 相反，c-myb反义寡聚体强烈抑制或完全消除T细胞白血病系的克隆形成生长，78%（23例中的18例）的原发性急性髓性白血病病例被检查， 5例原发性慢性粒细胞白血病（CML）急变中4例为白血病。 在后者的三个患者中，聚合酶链反应分析的c-myb反义处理的正常和CML细胞的1：1混合物显示bcr-abl表达的完全缺乏，这表明CML克隆形成单位已被完全消除的文化。 在抑制白血病细胞生长的反义剂量下，正常的造血祖细胞存活。 因此，正常和白血病造血细胞对c-myb反义DNA的毒性作用表现出不同的敏感性。 用反义寡脱氧核苷酸干扰c-myb功能可能最终形成白血病治疗的分子方法的基础，也许最直接的是作为离体骨髓净化剂。
The c-myb protooncogene is preferentially expressed in hematopoietic cells, and its encoded protein, Myb, is required for hematopoietic cell proliferation. To analyze the relative Myb dependence of normal and leukemic human hematopoietic progenitor cells, normal bone marrow cells, several types of leukemic blast cells, and 1:1 mixtures of normal and leukemic cells were cultured in the presence of c-myb sense or antisense oligodeoxynucleotides; cell viability and cloning efficiency were then assessed. c-myb sense oligomers had negligible effect on normal and leukemic cells. In contrast, c-myb antisense oligomers strongly inhibited or completely abolished clonogenic growth of a T-cell leukemia line, 78% (18 of 23) of primary acute myelogenous leukemia cases examined, and 4 of 5 primary chronic myelogenous leukemia (CML) cases in blast crisis. In three of the latter patients, polymerase chain reaction analysis of a 1:1 mixture of c-myb antisense-treated normal and CML cells revealed a complete absence of bcr-abl expression, suggesting that the CML clonogenic units had been completely eliminated from the cultures. At antisense doses that inhibited leukemic cell growth, normal hematopoietic progenitor cells survived. Thus, normal and leukemic hematopoietic cells show differential sensitivity to the toxic effects of c-myb antisense DNA. Perturbation of c-myb function with antisense oligodeoxynucleotides might eventually form the basis for a molecular approach to leukemia therapy, perhaps most immediately as ex vivo bone marrow purging agents.