Short Double-Stranded RNA with Immunostimulatory Activity: Sequence Dependence
Short Double-Stranded RNA with Immunostimulatory Activity: Sequence Dependence
复制标题
DOI：
10.1089/nat.2011.0328
复制
发表时间：
2012-06-01
影响因子：
4
通讯作者：
Chernolovskaya, Elena L.
中科院分区：
文献类型：
--
作者：
Kabilova, Tatyana O.;Meschaninova, Mariya I.;Chernolovskaya, Elena L.
小干扰rna （siRNA）能够激活哺乳动物的先天免疫系统，这取决于它们的结构、序列和传递方法。双链RNA的免疫刺激活性可应用于抗病毒和抗肿瘤治疗。在这里，我们鉴定了一组19 bp的RNA双链，在3'端有3个核苷悬垂，显示出免疫刺激活性（这里和免疫刺激RNA或isRNA之后），并研究了它们的序列/活性关系。研究发现，在isRNA序列的中间部分（从1链5‘端开始计数的10-16个位置）引入取代并不会改变其抗增殖活性，而在isRNA的3’端区域引入取代则会大大降低其抗增殖活性。isrna能有效抑制人口腔表皮样癌细胞的增殖[半最大抑制浓度（IC50）值在10 ~ 100 nM之间变化]。我们的研究表明，isrna的抗增殖作用与细胞生长阻滞有关，而不是诱导细胞凋亡。这些isrna强烈刺激干扰素- α (ifn - α)的合成，并在较小程度上刺激黏附的外周血单核细胞中肿瘤坏死因子（tnf - α）和白细胞介素-6 （IL-6）的合成。与对照小鼠相比，静脉注射isRNA/脂质体复合物使C57BL小鼠血清中的ifn - α和IL-6水平分别增加了15倍和3倍。所获得的结果清楚地表明所研究的isrna具有明显的免疫刺激和抗增殖特性。因此，这些短双链rna可以被认为是治疗肿瘤和病毒性疾病的潜在药物。
Small interfering RNAs (siRNA) are able to activate the mammalian innate immune system depending on their structure, sequence, and method of delivery. The immunostimulatory activity of double-stranded RNA can be applied to antiviral and antitumor therapy. Here we identified a set of 19-bp RNA duplexes with 3-nucleotid overhangs in the 3' ends that display immunostimulating activity (here and after immunostimulating RNA, or isRNA) and studied their sequence/activity relationships. It was found that the introduction of substitutions in the middle part of the isRNA sequence (10-16 positions counting from the 5' end of strand 1) does not alter the antiproliferative activity, while substitutions in the 3' end region of isRNA substantially reduce it. isRNAs efficiently inhibit the proliferation of human oral epidermoid carcinoma cells [half-maximal inhibitory concentration (IC50) values varied from 10 to 100 nM]. Our research demonstrated that antiproliferative effects of isRNAs are related to cell growth arrest, rather than the induction of apoptosis. These isRNAs strongly stimulate the synthesis of interferon-alpha (IFN-alpha), and to a lesser extent the synthesis of tumor necrosis factor (TNF-alpha) and interleukin-6 (IL-6), in adherent peripheral blood mononuclear cells. An intravenous injection of isRNA/Lipofectamine complexes into C57BL mice increases IFN-alpha and IL-6 levels in the blood serum up to 15-fold and 3-fold, respectively, compared to the control mice. The results obtained clearly demonstrate the pronounced immunostimulatory and antiproliferative properties of the isRNAs under study. Hence, these short double-stranded RNAs can be considered as potential agents for the therapy of oncological and viral diseases.