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为了探讨锤头状核酶对原代慢性粒细胞白血病 (chronicmyelogenousleukemia ,CML)细胞的作用及其在体外骨髓净化的效果 ,应用逆转录病毒介导的基因转移法将针对bcr abl融合基因的核酶转染原代CML和正常人骨髓单个核细胞 ,通过造血祖细胞集落培养、流式细胞术、免疫细胞化学法检测核酶对原代CML细胞的作用 ;进一步将此核酶转染CML缓解模型 ,通过白血病细胞集落培养、巢式RT PCR检测其在体外骨髓净化的效果。结果表明 ,原代CML细胞生长增殖和p2 10的表达显著受抑 ,但正常造血祖细胞受影响较小 ;核酶完全抑制模拟缓解骨髓中残留K5 6 2细胞的增殖及bcr ablmRNA表达 ,但并不影响ablmRNA表达 ,因此有望用于CML的体外骨髓净化。
In order to investigate the effect of hammerhead ribozyme on primary myelogenous leukemia (CML) cells and its effect on bone marrow purification in vitro, retrovirus-mediated gene transfer was used to clone ribozymes targeting bcr abl fusion gene Transfection of primary CML and normal human bone marrow mononuclear cells, hematopoietic progenitor cell colony culture, flow cytometry, immunocytochemistry detection of ribozyme on the primary CML cells; further transfer of this ribozyme CML remission model , Through leukemia cell colony culture, nested RT PCR detection of its in vitro bone marrow purification effect. The results showed that the proliferation of primary CML cells and the expression of p2 10 were significantly inhibited, but the normal hematopoietic progenitor cells were less affected. Ribozymes completely inhibited the mimicry to alleviate the proliferation and bcr abl mRNA expression in residual K5 6 2 cells Does not affect ablmRNA expression, and is therefore expected to be used for in vitro bone marrow purification of CML.