Nanoparticle-based delivery of siDCAMKL-1 increases microRNA-144 and inhibits colorectal cancer tumor growth via a Notch-1 dependent mechanism.
Nanoparticle-based delivery of siDCAMKL-1 increases microRNA-144 and inhibits colorectal cancer tumor growth via a Notch-1 dependent mechanism.
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DOI：
10.1186/1477-3155-9-40
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发表时间：
2011-09-19
影响因子：
10.2
通讯作者：
Houchen CW
中科院分区：
文献类型：
--
作者：
Sureban SM;May R;Mondalek FG;Qu D;Ponnurangam S;Pantazis P;Anant S;Ramanujam RP;Houchen CW
能够运输小干扰RNA (siRNA) 的有效药物递送系统的开发一直难以实现。我们之前曾报道过，用 siDCAMKL-1 脂质体制剂治疗后，结直肠癌肿瘤异种移植物的生长被抑制。在本报告中，我们利用纳米颗粒 (NP) 技术来传递 DCAMKL-1 特异性 siRNA，以击倒潜在的关键癌症调节因子。在本研究中，使用实时 RT-PCR 分析 mRNA/miRNA，并通过蛋白质印迹/免疫组织化学分析蛋白质。 siDCAMKL-1 封装在聚丙交酯乙交酯共聚物 (NP-siDCAMKL-1) 中；在裸鼠中产生肿瘤异种移植物，并用 NP-siDCAMKL-1 和 DAPT（γ-分泌酶抑制剂）单独或联合治疗。为了在体外测量let-7a和miR-144表达，用编码萤火虫荧光素酶基因的质粒转染HCT116细胞，该基因在3'UTR中具有let-7a和miR-144 miRNA结合位点。将 NP-siDCAMKL-1 注入 HCT116 异种移植物中，分别通过 let-7a 和 miR-144 miRNA 依赖性机制导致肿瘤生长停滞、原癌基因 c-Myc 和 Notch-1 下调。体外观察到 let-7a 和 miR-144 特异性荧光素酶活性相应降低。此外，在体内观察到 EMT 抑制剂 miR-200a 的上调和 EMT 相关转录因子 ZEB1、ZEB2、Snail 和 Slug 的下调。最后，DAPT 介导的 Notch-1 抑制导致 HCT116 肿瘤生长停滞，并通过 miR-144 依赖性机制下调 Notch-1。这些发现表明，针对 DCAMKL-1 等关键靶标的基于纳米颗粒的 siRNA 递送可能提供一种通过调节内源 miRNA 来治疗癌症的新方法。
The development of effective drug delivery systems capable of transporting small interfering RNA (siRNA) has been elusive. We have previously reported that colorectal cancer tumor xenograft growth was arrested following treatment with liposomal preparation of siDCAMKL-1. In this report, we have utilized Nanoparticle (NP) technology to deliver DCAMKL-1 specific siRNA to knockdown potential key cancer regulators. In this study, mRNA/miRNA were analyzed using real-time RT-PCR and protein by western blot/immunohistochemistry. siDCAMKL-1 was encapsulated in Poly(lactide-co-glycolide)-based NPs (NP-siDCAMKL-1); Tumor xenografts were generated in nude mice, treated with NP-siDCAMKL-1 and DAPT (γ-secretase inhibitor) alone and in combination. To measure let-7a and miR-144 expression in vitro, HCT116 cells were transfected with plasmids encoding the firefly luciferase gene with let-7a and miR-144 miRNA binding sites in the 3'UTR. Administration of NP-siDCAMKL-1 into HCT116 xenografts resulted in tumor growth arrest, downregulation of proto-oncogene c-Myc and Notch-1 via let-7a and miR-144 miRNA-dependent mechanisms, respectively. A corresponding reduction in let-7a and miR-144 specific luciferase activity was observed in vitro. Moreover, an upregulation of EMT inhibitor miR-200a and downregulation of the EMT-associated transcription factors ZEB1, ZEB2, Snail and Slug were observed in vivo. Lastly, DAPT-mediated inhibition of Notch-1 resulted in HCT116 tumor growth arrest and down regulation of Notch-1 via a miR-144 dependent mechanism. These findings demonstrate that nanoparticle-based delivery of siRNAs directed at critical targets such as DCAMKL-1 may provide a novel approach to treat cancer through the regulation of endogenous miRNAs.
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影响因子：
254.7
作者：
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通讯作者：
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影响因子：
5.2
作者：
May, Randal;Sureban, Sripathi M.;Hoang, Nguyet;Riehl, Terrence E.;Lightfoot, Stan A.;Ramanujam, Rama;Wyche, James H.;Anant, Shrikant;Houchen, Courtney W.
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具有靶向潜力的聚合物造影剂。
影响因子：
4.2
作者：
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通讯作者：
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影响因子：
8
作者：
Leethanakul, C;Patel, V;Gutkind, JS
通讯作者：
Gutkind, JS
影响因子：
5.2
作者：
May, Randal;Riehl, Terrence E.;Houchen, Courtney W.
通讯作者：
Houchen, Courtney W.