Citro G, Perrotti D, Cucco C, D'Agnano I, Sacchi A, Zupi G, Calabretta B
Laboratorio Chemioterapia Sperimentale, Istituto Tumori Regina Elena, Rome, Italy.
Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7031-5. doi: 10.1073/pnas.89.15.7031.
Exposure of human leukemia HL-60 cells to an oligodeoxynucleotide complementary to an 18-base sequence (codons 2-7) of c-myb-encoded mRNA has previously been shown to result in inhibition of cell proliferation. Because HL-60 cells express high levels of transferrin receptor we adapted a DNA delivery system based on receptor-mediated endocytosis to introduce myb oligomers complexed with a transferrin-polylysine conjugate into those cells. A DNA.RNA duplex resistant to S1 nuclease digestion was detected as early as 12 hr after culture of HL-60 cells in the presence of the myb antisense/transferrin-polylysine complex. Exposure of HL-60 cells to the myb antisense/transferrin-polylysine complex resulted in rapid and profound inhibition of proliferation and loss of cell viability much more pronounced than that occurring in cells exposed to free myb antisense oligodeoxynucleotides. The transferrin-polylysine/myb sense complex or the transferrin-polylysine conjugate alone had no effect on HL-60 cell proliferation and viability. These findings indicate that myb synthetic oligodeoxynucleotides enter efficiently into HL-60 by transferrin receptor-mediated endocytosis and exert a profound biological effect. Such a delivery system could exploit other ligand-receptor interactions for the selective delivery of oncogene-targeted antisense oligodeoxynucleotides.
先前的研究表明,将人白血病HL-60细胞暴露于与c-myb编码的mRNA的18个碱基序列(密码子2-7)互补的寡聚脱氧核苷酸中,会导致细胞增殖受到抑制。由于HL-60细胞表达高水平的转铁蛋白受体,我们采用了一种基于受体介导的内吞作用的DNA递送系统,将与转铁蛋白-聚赖氨酸缀合物复合的myb寡聚物导入这些细胞。早在HL-60细胞在myb反义/转铁蛋白-聚赖氨酸复合物存在下培养12小时后,就检测到了对S1核酸酶消化具有抗性的DNA.RNA双链体。将HL-60细胞暴露于myb反义/转铁蛋白-聚赖氨酸复合物中,会导致增殖迅速且显著受到抑制,细胞活力丧失,这比暴露于游离myb反义寡聚脱氧核苷酸的细胞中发生的情况更为明显。转铁蛋白-聚赖氨酸/myb正义复合物或单独的转铁蛋白-聚赖氨酸缀合物对HL-60细胞的增殖和活力没有影响。这些发现表明,myb合成寡聚脱氧核苷酸通过转铁蛋白受体介导的内吞作用有效地进入HL-60细胞,并发挥深远的生物学效应。这样的递送系统可以利用其他配体-受体相互作用来选择性递送针对癌基因的反义寡聚脱氧核苷酸。