Transformed and immortalized cellular uptake of oligodeoxynucleoside phosphorothioates, 3'-alkylamino oligodeoxynucleotides, 2'-O-methyl oligoribonucleotides, oligodeoxynucleoside methylphosphonates, and peptide nucleic acids
Transformed and immortalized cellular uptake of oligodeoxynucleoside phosphorothioates, 3'-alkylamino oligodeoxynucleotides, 2'-O-methyl oligoribonucleotides, oligodeoxynucleoside methylphosphonates, and peptide nucleic acids
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DOI：
10.1016/s0006-2952(97)82440-9
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发表时间：
1997-05-15
影响因子：
5.8
通讯作者：
Wickstrom, E
中科院分区：
文献类型：
--
作者：
Gray, GD;Basu, S;Wickstrom, E
在各种各样的类似物和细胞类型的细胞摄取的直接定量比较是必要的寡核苷酸诊断和治疗应用的设计。本工作报告了几种转化或永生化细胞系中[C-14]寡脱氧核苷硫代磷酸酯（PS）、3 '-烷基氨基寡脱氧核苷磷酸二酯（PO-NH 2）、2'-O-甲基寡核糖核苷磷酸二酯（2 OM）、肽核酸（PNA）和寡脱氧核苷甲基膦酸酯（MP）的定量细胞摄取和核定位。所有类似物都表现出活跃的细胞摄取，在1-3小时内细胞内浓度大大超过细胞外1 μ M浓度。然而，到9-24小时，PS的细胞积累超过PO-NH 2和2 OM的3- 5倍，PNA的6- 7倍，MP的8- 10倍。在两个转化细胞系，HL 60白细胞和H-ras转化成纤维细胞，使用三种不同的异质序列观察到类似的结果。H-ras和IGF-1 R转化的成纤维细胞对所有类似物的摄取比非转化的永生化成纤维细胞高2至5倍。PO-NH 2，PS和MP类似物的核水平约为总细胞摄取的25%，而2 OM和PNA的核百分比小于20%，表明类似物之间的核定位存在一些差异。这些观察结果提供了作为寡核苷酸修饰的函数的细胞摄取的直接定量比较，并且暗示转化增强细胞摄取。从治疗和诊断的角度来看，在一方面的摄取效率与另一方面的核酸酶抗性和杂交强度之间存在明显的权衡。(C)1997年爱思唯尔科学公司
Direct quantitative comparisons of cellular uptake across a wide variety of analogs and cell types are necessary for the design of oligonucleotide diagnostic and therapeutic applications. This work reports quantitative cellular uptake and nuclear localization of [C-14]oligodeoxynucleoside phosphorothioates (PS), 3'-alkylamino oligodeoxynucleoside phosphodiesters (PO-NH2), 2'-O-methyl oligoribonucleoside phosphodiesters (2OM), peptide nucleic acids (PNA), and oligodeoxynucleoside methylphosphonates (MP) in several transformed or immortalized cell lines. All analogs demonstrated active cellular uptake in that intracellular concentrations greatly exceeded the extracellular 1 mu M concentration within 1-3 hr. However, by 9-24 hr, cellular accumulations of PS exceeded those of PO-NH2 and 2OM by 3- to 5-fold, PNA by 6- to 7-fold, and MP by 8- to 10-fold. Similar results were observed in two transformed cell lines, HL 60 leukocytes and H-ras transformed fibroblasts, using three different heterogeneous sequences. H-ras and IGF-1R transformed fibroblasts had a 2- to 5-fold higher uptake of all analogs than non-transformed immortalized fibroblasts. Nuclear levels of the PO-NH2, PS, and MP analogs were approximately 25% of total cellular uptake, while nuclear percentages of 2OM and PNA were less than 20%, suggesting some differences in nuclear localization among the analogs. These observations provide a direct quantitative comparison of cellular uptake as a function of oligonucleotide modification, and imply that transformation enhances cellular uptake. From the perspective of therapy and diagnosis, clear trade-offs were apparent between efficiency of uptake on the one hand, and nuclease resistance and hybridization strength on the other. (C) 1997 Elsevier Science Inc.