Cationized gelatin delivery of a plasmid DNA expressing small interference RNA for VEGF inhibits murine squamous cell carcinoma
Cationized gelatin delivery of a plasmid DNA expressing small interference RNA for VEGF inhibits murine squamous cell carcinoma
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DOI：
10.1111/j.1349-7006.2006.00174.x
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发表时间：
2006-04-01
期刊：
影响因子：
5.7
通讯作者：
Tabata, Y
中科院分区：
文献类型：
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作者：
Matsumoto, G;Kushibiki, T;Tabata, Y
双链RNA（dsRNA）在RNA干扰（RNAi）中起主要作用，在RNAi中，dsRNA的片段最初被Dicer切割成称为小干扰RNA（siRNA）的较短片段（21-23 nt）。然后，这些siRNA特异性靶向同源mRNA分子，使它们被细胞核糖核酸酶降解。RNAi下调哺乳动物细胞中的内源基因表达。血管内皮生长因子（VEGF）是血管发生和血管生成的关键分子。肿瘤生长是一个血管生成依赖的过程，旨在抑制血管生成的治疗策略在理论上是有吸引力的。为了研究在VEGF mRNA的特异性敲低中使用VEGF的siRNA的可行性，从而抑制血管生成，我们已经用基于靶向VEGF的siRNA系统（siVEGF）的DNA载体进行了实验。它几乎完全抑制小鼠鳞状细胞癌NRS-1细胞中VEGF mRNA的三种不同亚型（VEGF 120、VEGF 164和VEGF 188）的表达和VEGF蛋白的分泌。从阳离子化明胶微球释放的siVEGF在体内抑制肿瘤生长。血管分布的显著减少伴随着siVEGF转染的肿瘤的抑制。荧光显微镜观察显示，注射后10天，肿瘤周围仍可见siVEGF与阳离子明胶微球的复合物，而游离的siVEGF已消失。siVEGF基因治疗增加了周细胞覆盖的血管比例，并诱导周细胞表达血管生成素-1。这些数据表明，含有siVEGF的阳离子化明胶微球可用于在NRS-1鳞状细胞癌异种移植模型中使肿瘤血管系统正常化并抑制肿瘤生长。
Double-stranded RNA (dsRNA) plays a major role in RNA interference (RNAi), a process in which segments of dsRNA are initially cleaved by the Dicer into shorter segments (21-23 nt) called small interfering RNA (siRNA). These siRNA then specifically target homologous mRNA molecules causing them to be degraded by cellular ribonucleases. RNAi downregulates endogenous gene expression in mammalian cells. Vascular endothelial growth factor (VEGF) is a key molecule in vasculogenesis as well as in angiogenesis. Tumor growth is an angiogenesis-dependent process, and therapeutic strategies aimed at inhibiting angiogenesis are theoretically attractive. To investigate the feasibility of using siRNA for VEGF in the specific knockdown of VEGF mRNA, thereby inhibiting angiogenesis, we have performed experiments with a DNA vector based on a siRNA system that targets VEGF (siVEGF). It almost completely inhibited the expression of three different isoforms (VEGF120, VEGF164 and VEGF188) of VEGF mRNA and the secretion of VEGF protein in mouse squamous cell carcinoma NRS-1 cells. The siVEGF released from cationized gelatin microspheres suppressed tumor growth in vivo. A marked reduction in vascularity accompanied the inhibition of a siVEGF-transfected tumor. Fluorescent microscopic study showed that the complex of siVEGF with cationized gelatin microspheres was still present around the tumor 10 days after injection, while free siVEGF had vanished by that time. siVEGF gene therapy increased the fraction of vessels covered by pericytes and induced expression of angiopoietin-1 by pericytes. These data suggest that cationized-gelatin microspheres containing siVEGF can be used to normalize tumor vasculature and inhibit tumor growth in a NRS-1 squamous cell carcinoma xenograft model.