Delivery of CD44 shRNA/Nanoparticles within Cancer Cells PERTURBATION OF HYALURONAN/CD44v6 INTERACTIONS AND REDUCTION IN ADENOMA GROWTH IN Apc Min/ plus MICE
Delivery of CD44 shRNA/Nanoparticles within Cancer Cells PERTURBATION OF HYALURONAN/CD44v6 INTERACTIONS AND REDUCTION IN ADENOMA GROWTH IN Apc Min/ plus MICE
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DOI：
10.1074/jbc.m806772200
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发表时间：
2009-05-01
影响因子：
4.8
通讯作者：
Ghatak, Shibnath
中科院分区：
文献类型：
--
作者：
Misra, Suniti;Hascall, Vincent C.;Ghatak, Shibnath
我们的研究表明，透明质酸与CD44在肿瘤细胞上的结构性相互作用通过形成含有激活的受体酪氨酸激酶的多聚体信号复合体来诱导多种抗细胞凋亡的细胞生存途径。透明质酸-CD44相互作用拮抗剂抑制肿瘤细胞上的透明质酸-CD44相互作用，通过分解复合体来抑制这些活性。尽管透明质酸-CD44相互作用拮抗剂--透明质酸-低聚糖的抗肿瘤活性能有效地增强肿瘤细胞对化疗药物的敏感性，并抑制移植瘤的生长，但单用透明质酸-低聚糖不能有效地抑制APC Min/+小鼠的肿瘤进展。我们现在在体外和体内证明，靶向抑制CD44v6的表达会耗尽结肠肿瘤细胞通过透明质酸-CD44v6相互作用发出信号的能力。首先，我们将编码CD44v6 shRNA的寡核苷酸克隆到条件沉默的pSICO载体中。其次，利用pSICO-CD44v6 shRNA和结肠特异性Fabp1启动子驱动的Cre重组酶表达载体包装成转铁蛋白包裹的纳米颗粒，成功地将CD44v6 shRNA导入癌前病变和肿瘤癌变的结肠癌细胞中。第三，利用APC Min/+小鼠模型，我们证明了抑制CD44v6的表达减少了通过透明质酸/CD44v6-pErbB2-COX-2相互作用的信号通路，从而减少了腺瘤的数量和生长。总之，这些数据为短发夹状RNA/纳米颗粒技术的新治疗策略及其沉默与结肠肿瘤细胞相关基因的潜力提供了洞察力。
Our studies have shown that constitutive interactions between hyaluronan and CD44 on tumor cells induces various anti-apoptotic cell survival pathways through the formation of a multimeric signaling complex that contains activated receptor tyrosine kinases. Inhibition of the hyaluronan-CD44 interactions on tumor cells by hyaluronan-CD44 interaction antagonists suppresses these activities by disassembling the complex. Although the anti-tumor activity of hyaluronan-oligosaccharides, a hyaluronan-CD44 interaction antagonist, is effective in sensitizing tumor cells to chemotherapeutic agents and reducing tumor growth in xenografts, hyaluronan-oligosaccharide alone was not effective in reducing tumor progression in Apc Min/+ mice. We now show in vitro and in vivo that targeted inhibition of the expression of CD44v6 depletes the ability of the colon tumor cells to signal through hyaluronan-CD44v6 interactions. First, we cloned oligonucleotides coding CD44v6 shRNA into a conditionally silenced pSico vector. Second, using pSico-CD44v6 shRNA and a colon-specific Fabpl promoter-driven Cre recombinase expression vector packaged into transferrin-coated nanoparticles, we successfully delivered the CD44v6 shRNA within pre-neoplastic and neoplastic colon malignant cells. Third, using the Apc Min/+ mice model, we demonstrated that inhibition of the CD44v6 expression reduces the signaling through a hyaluronan/CD44v6-pErbB2-Cox-2 interaction pathway and reduced adenoma number and growth. Together, these data provide insight into the novel therapeutic strategies of short hairpin RNA/nanoparticle technology and its potential for silencing genes associated with colon tumor cells.