Aerosol delivery of glucosylated polyethylenimine/phosphatase and tensin homologue deleted on chromosome 10 complex suppresses Akt downstream pathways in the lung of K-ras null mice
Aerosol delivery of glucosylated polyethylenimine/phosphatase and tensin homologue deleted on chromosome 10 complex suppresses Akt downstream pathways in the lung of K-ras null mice
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DOI：
10.1158/0008-5472.can-04-1231
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发表时间：
2004-11-01
期刊：
影响因子：
11.2
通讯作者：
Cho, MH
中科院分区：
文献类型：
--
作者：
Kim, HW;Park, IK;Cho, MH
传统疗法治疗肺癌患者长期生存的困难表明需要新的方法。尽管已经研究了使用基因递送的几种基因的抗肿瘤活性，但是围绕所使用的方法的问题，例如效率、特异性和毒性，阻碍了其作为有效疗法的应用。这使得气溶胶基因递送作为一种非侵入性的肺癌治疗方法重新出现。在这项研究中，葡萄糖基化的共轭聚乙烯亚胺（葡萄糖基化PEI）被用作载体。在确认了肺中葡糖基化PEI载体的效率后，研究了10号染色体上缺失的磷酸酶和张力蛋白同源物（PTEN）肿瘤抑制基因对Akt下游途径的潜在影响。将含有葡萄糖基化PEI和重组质粒pcDNA3.0-PTEN复合物的气雾剂通过鼻吸入系统递送到K-ras基因敲除肺癌模型小鼠体内。对经PTEN转染的小鼠肺组织中磷脂酰肌醇3 '-激酶/Akt信号通路中蛋白质的研究显示，PTEN蛋白高度表达，而PDK 1、总Akt 1、磷酸化-（Thr-308）-Akt、磷酸化-（Ser-2448）-mTOR、p70 S6 K和4 E-BP 1的蛋白质水平不同程度地降低。此外，Akt和mTOR的激酶活性均被抑制。最后，通过末端脱氧核苷酸转移酶介导的缺口末端标记法在PTEN递送的小鼠肺中检测到细胞凋亡，这表明我们的气溶胶PTEN递送能够在体内功能性地改变细胞表型。总之，蛋白质印迹分析，激酶测定，免疫组化，和末端脱氧核苷酸转移酶介导的缺口末端标记试验表明，我们的气溶胶基因递送技术是与体内基因递送兼容，可以作为一种非侵入性的基因治疗。
Difficulties in achieving long-term survival of lung cancer patients treated with conventional therapies suggest that novel approaches are required. Although several genes have been investigated for antitumor activities using gene delivery, problems surrounding the methods used such as efficiency, specificity, and toxicity hinder its application as an effective therapy. This has lead to the re-emergence of aerosol gene delivery as a noninvasive approach to lung cancer therapy. In this study, glucosylated conjugated polyethylenimine (glucosylated PEI) was used as carrier. After confirming the efficiency of glucosylated PEI carriers in lungs, the potential effects of the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) tumor suppressor gene on Akt downstream pathways were investigated. Aerosol containing glucosylated PEI and recombinant plasmid pcDNA3.0-PTEN complex was delivered into K-ras null lung cancer model mice through a nose-only inhalation system. Investigation of proteins in the phosphatidylinositol 3'-kinase/Akt signaling pathway in PTEN-delivered mouse lung revealed that the PTEN protein was highly expressed, whereas the protein levels of PDK1, total Akt1, phospho-(Thr-308)-Akt, phospho-(Ser-2448)-mTOR, p70S6K, and 4E-BP1 were decreased to varying degrees. Additionally, the kinase activities of both Akt and mTOR were suppressed. Finally, apoptosis was detected in PTEN-delivered mouse lung by terminal deoxynucleotidyl-transferase-mediated nick end labeling assay, suggesting that our aerosol PTEN delivery is capable of functionally altering cell phenotype ill vivo. In summary, Western blot analysis, kinase assays, immunohistochemistry, and terminal deoxynucleotidyltransferase-mediated nick end labeling assays suggest that our aerosol gene delivery technique is compatible with ill vivo gene delivery and can be applied as a noninvasive gene therapy.