TRANSFERRIN POLYCATION-MEDIATED INTRODUCTION OF DNA INTO HUMAN LEUKEMIC-CELLS - STIMULATION BY AGENTS THAT AFFECT THE SURVIVAL OF TRANSFECTED DNA OR MODULATE TRANSFERRIN RECEPTOR LEVELS
TRANSFERRIN POLYCATION-MEDIATED INTRODUCTION OF DNA INTO HUMAN LEUKEMIC-CELLS - STIMULATION BY AGENTS THAT AFFECT THE SURVIVAL OF TRANSFECTED DNA OR MODULATE TRANSFERRIN RECEPTOR LEVELS
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DOI：
10.1073/pnas.87.11.4033
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发表时间：
1990-06-01
影响因子：
11.1
通讯作者：
BIRNSTIEL, ML
中科院分区：
文献类型：
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作者：
COTTEN, M;LANGLEROUAULT, F;BIRNSTIEL, ML
我们已经破坏了受体介导的内吞事件，将基因转运到人类白血病细胞中。通过将天然铁传递蛋白转铁蛋白与dna结合多阳离子聚赖氨酸或鱼精蛋白偶联，我们创造了结合核酸并在正常转铁蛋白周期中将其带入细胞的蛋白质偶联物[Wagner， E., Zenke， M., Cotten， M., Beug, H . and Birnstiel， M.L. (1990) Proc. Natl。学会科学。[j]。我们在这里证明了这种方法对人类白血病细胞系是有用的。我们通过(i)用细胞渗透性铁螯合剂去铁胺处理细胞增加转铁蛋白受体密度，（ii）用琥珀酰丙酮处理干扰血红素的合成，或（iii）用氯化钴处理刺激血红素的降解来提高基因传递率。与作为内吞事件的基因传递一致，我们表明，随后在K-562细胞中，包含在运输DNA中的基因的表达取决于细胞中溶酶性收缩剂氯喹的存在。相比之下，莫能菌素阻断了“转移感染”，在18度的温度下培养细胞也是如此。C。
We have subverted a receptor-mediated endocytosis event to transport genes into human leukemic cells. By coupling the natural iron-delivery protein transferrin to the DNA-binding polycations polylysine or protamine, we have created protein conjugates that bind nucleic acids and carry them into the cell during the normal transferrin cycle [Wagner, E., Zenke, M., Cotten, M., Beug, H and Birnstiel, M.L. (1990) Proc. Natl. Acad. Sci. USA 87, 3410-3414]. We demonstrate here that this procedure is useful for a human leukemic cell line. We enhanced the rate of gene delivery by (i) increasing the transferrin receptor density through treatment of the cells with the cell-permeable iron chelator desferrioxamine, (ii) interfering with the synthesis of heme with succinyl acetone treatment, or (iii) stimulating the degradation of heme with cobalt chloride treatment. Consistent with gene delivery as an endocytosis event, we show that the subsequent expression in K-562 cells of a gene included in the transported DNA depends upon the cellular presence of the lysosomotropic agent chloroquine. By contrast, monensin blocks "transferrinfection", as does incubation of the cells at 18.degree. C.