Antisense c-myc oligodeoxyribonucleotide cellular uptake.
Antisense c-myc oligodeoxyribonucleotide cellular uptake.
复制标题
反义c-myc寡脱氧核糖核苷酸细胞摄取。
DOI：
10.1023/a:1015846209681
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发表时间：
1992
影响因子：
3.7
通讯作者：
Hunt,CA
中科院分区：
文献类型：
--
作者：
Wu-Pong,S;Weiss,TL;Hunt,CA
反义寡核苷酸作为基因表达抑制剂具有治疗潜力。然而，完整的寡核苷酸到达细胞内作用位点的机制尚不清楚。在这项研究中，我们使用的寡脱氧核糖核苷酸21聚体互补的c-mycproto-癌基因的翻译起始密码子，研究寡核苷酸的摄取和内化到Rauscher Red 5-1.5细胞的机制。我们发现胰蛋白酶敏感和胰蛋白酶不敏感的表面结合，除了内化。摄取是部分能量依赖性的，并被带电分子抑制，包括DNA、ATP、随机序列寡核苷酸和硫酸葡聚糖。摄取似乎不发生通过传统的受体介导的摄取途径，因为氯喹，莫能菌素，氧化苯胂预处理不显着降低内化。阴离子通道抑制剂SITS和盐NaCl、Na 2SO 4和NH 4Cl显著降低寡核苷酸摄取。摄取是否通过通道或新的摄取机制发生仍然是未知的。提出了一个模型，合理地模拟实验数据。
Antisense oligonucleotides have therapeutic potential as inhibitors of gene expression. However, the mechanism by which an intact oligonucleotide reaches the intracellular site of action is unknown. In this study, we use an Oligodeoxyribonucleotide 21-mer complementary to the translation initiation codon of the c-mycproto-oncogene to study the mechanism of oligonucleotide uptake and internalization into Rauscher Red 5-1.5 cells. We find trypsin-sensitive and trypsin-insensitive surface binding, in addition to internalization. Uptake is partially energy dependent and inhibited by charged molecules, including DNA, ATP, a random sequence oligonucleotide, and dextran sulfate. Uptake does not appear to occur via a traditional receptor-mediated uptake pathway because chloro-quine, monensin, and phenylarsine oxide pretreatment does not significantly decrease internalization. An anion channel inhibitor, SITS, and the salts, NaCl, Na2SO4, and NH4Cl, significantly decrease oligonucleotide uptake. Whether uptake occurs via a channel or a novel uptake mechanism is still unknown. A model is proposed which reasonably simulates the experimental data.