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荷兰应用科学研究院放射生物学研究所建立的可移植性大鼠急性白血病(brown Norway rat acutemyelocytic leukemia,BNML)是研究人急性粒系白血病的颇为完备的实验模型。由于重组人白细胞介素-6(rhIL-6)可作用于大鼠,然而大鼠BNML细胞系LT12缺乏rhIL-6受体(rhIL—6R)表达。为了在BNML模型上探讨rhIL-6在体外以及体内对白血病细胞增殖和分化的影响,并探讨细胞因子-毒素联接物用于白血病靶向治疗的可能性,本研究应用改良的磷酸钙共沉淀法将携带rhIL-6R cDNA的逆转录病毒表达质粒XM6/IL—6RNeo导入LT12细胞,经过G418筛选获得G418抗性克隆,建立了表达rhIL—6R的LT12亚细胞
Transplantation of rat brown Norway rat acutemyelocytic leukemia (BNML) established by the Institute of Radiobiology of the Netherlands Institute for Applied Science is a well-established experimental model for the study of human acute myeloid leukemia. Since recombinant human interleukin-6 (rhIL-6) acts in rats, rat BNML cell line LT12 lacks rhIL-6 receptor (rhIL-6R) expression. In order to investigate the effect of rhIL-6 on the proliferation and differentiation of leukemia cells in vitro and in vivo and to explore the possibility of cytokine-toxin conjugate for targeted therapy of leukemia in BNML model, this study applied modified calcium phosphate co-precipitation method The recombinant retroviral vector XM6 / IL-6RNeo carrying rhIL-6R cDNA was introduced into LT12 cells and G418-resistant clones were screened by G418 to establish the rhIL-6R-expressing LT12 subunit