Small interfering RNAs induce macrophage migration inhibitory factor production and proliferation in breast cancer cells via a double-stranded RNA-dependent protein kinase-dependent mechanism
Small interfering RNAs induce macrophage migration inhibitory factor production and proliferation in breast cancer cells via a double-stranded RNA-dependent protein kinase-dependent mechanism
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DOI：
10.4049/jimmunol.180.11.7125
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发表时间：
2008-06-01
影响因子：
4.4
通讯作者：
Donnelly, Seamas C.
中科院分区：
文献类型：
--
作者：
Armstrong, Michelle E.;Gantier, Michael;Donnelly, Seamas C.
小干扰RNA（siRNA）代表了在哺乳动物细胞中诱导基因沉默的新工具，并且目前正在进行临床试验以评估siRNA在各种人类疾病（包括年龄相关性黄斑变性和呼吸道合胞病毒感染）中的治疗功效。然而，先前报道的siRNA的脱靶、非特异性作用，包括I型IFN和促炎细胞因子的激活，仍然是关于这些药剂在体内使用的突出问题。巨噬细胞移动抑制因子（MIF）是一种多效性细胞因子，在细胞增殖、肿瘤发生和血管生成中具有良好的作用，并且是基于siRNA治疗乳腺癌的靶基因。然而，在这项研究中，我们描述了增加MIF生产乳腺癌（MCF-7）细胞转染后，与MIF siRNA和各种控制siRNA。这种作用被证明是剂量依赖性的，并在双链RNA依赖性蛋白激酶抑制剂2-氨基嘌呤的存在下减弱。此外，用poly（I：C）处理MCF-7细胞也刺激MCF-7细胞产生的MIF的PKR依赖性增加。siRNA诱导MCF-7细胞产生的MIF增加的生物学后果是乳腺癌细胞增殖的PKR依赖性增加。此外，在从原发性人乳腺癌群组制备的cDNA中;我们证明了PKR和MIF-mRNA表达之间的显著相关性（斯皮尔曼等级相关系数，r = 0.50，p < 0.0001，n = 63）。总之，这项研究强调了siRNAs的脱靶、非特异性作用的潜在生物学后果，并强调了使用siRNAs治疗人类疾病（如癌症）的安全性问题。
Small interfering RNAs (siRNAs) represent a novel tool to induce gene silencing in mammalian cells and clinical trials are currently ongoing to assess the therapeutic efficacy of siRNAs in various human diseases, including age-related macular degeneration and respiratory syncytial virus infection. However, previously reported off-target, nonspecific effects of siRNAs, including activation of type I IFNs and proinflammatory cytokines, remain an outstanding concern regarding use of these agents in vivo. Macrophage-migration inhibitory factor (MIF) is a pleiotropic cytokine with well-described roles in cell proliferation, tumorigenesis, and angiogenesis and represents a target gene for siRNA-based therapy in the treatment of breast cancer. However, in this study we describe an increase in MIF production from mammary adenocarcinoma (MCF-7) cells following transfection with MIF siRNA and various control siRNAs. This effect was shown to be dose-dependent and was attenuated in the presence of a double-stranded RNA-dependent protein kinase inhibitor, 2-aminopurine. Furthermore, treatment of MCF-7 cells with poly(I:C) also stimulated a PKR-dependent increase in MIF production from MCF-7 cells. The biological consequence of the siRNA-induced increase in MIF production from MCF-7 cells was a PKR-dependent increase in proliferation of breast cancer cells. Furthermore, in cDNAs prepared from a primary human breast cancer cohort;. we demonstrated a significant correlation (Spearman rank correlation coefficient, r = 0.50, p < 0.0001, n = 63) between PKR- and MIF-mRNA expression. in conclusion, this study highlights the potential biological consequences of off-target, nonspecific effects of siRNAs and underlines the safety concerns regarding the use of siRNAs in the treatment of human diseases, such as cancer.