Inhibition of cell proliferation and migration through nucleobase-modified polyamidoamine-mediated p53 delivery.
Inhibition of cell proliferation and migration through nucleobase-modified polyamidoamine-mediated p53 delivery.
复制标题
通过核碱基修饰的聚酰胺胺介导的 p53 递送抑制细胞增殖和迁移
DOI：
10.2147/ijn.s146917
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发表时间：
2018
影响因子：
8
通讯作者：
Li K
中科院分区：
文献类型：
--
作者：
Han H;Chen W;Yang J;Liang X;Wang Y;Li Q;Yang Y;Li K
前言：利用碱基2-氨基-6-氯嘌呤修饰的聚酰胺胺(AP-PAMAM)作为p53基因的载体，以达到抗肿瘤的目的。方法与材料用凝胶阻滞法检测pGFP-N3和pGL-3表达载体对pGFP-N3和PGL-3表达的影响。以人宫颈癌细胞系HeLa为模型，通过流式细胞仪、伤口愈合和Transwell迁移实验研究其对细胞增殖和迁移的抑制作用。用定量聚合酶链式反应和Western blotting方法检测P53的表达水平。结果在N/P比为2.0时，载体可将p5 3基因表达载体凝聚成稳定的纳米颗粒，且在所有N/P比下均可获得比聚酰胺胺(PAMAM)更高的转染率。AP-PAMAM介导的P53转导比PAMAM/P53具有更强的抗增殖作用。抗增殖作用是通过诱导细胞凋亡(凋亡率为26.17%)和细胞周期停滞于S期而触发的。此外，AP-PAMAM/P53的转导被发现能够抑制癌细胞的迁移和侵袭。最后，检测到P53基因在基因和蛋白水平的表达增强。结论PAMAM衍生物介导的P53基因治疗有望成为肿瘤基因治疗的一种新途径。
Introduction The nucleobase 2-amino-6-chloropurine-modified polyamidoamine (AP-PAMAM) was used as a carrier for p53 gene delivery to achieve the antitumor effects. Methods and materials The condensation of p53 plasmid was studied through gel retardation assay, and the transfection efficiency was evaluated through the transfection assay of pEGFP-N3 and pGL-3 plasmids. Using human cervical carcinoma cell line HeLa as a model, the inhibition of cell proliferation and migration was studied through flow cytometry, wound healing and Transwell migration assays, respectively. The p53 expression level was detected through quantitative polymerase chain reaction and Western blotting analyses. Results The carrier could condense p53 plasmid into stable nanoparticles at N/P ratios of 2.0, and higher transfection efficiency than polyamidoamine (PAMAM) could be obtained at all the N/P ratios studied. AP-PAMAM-mediated p53 delivery could achieve stronger antiproliferative effect than PAMAM/p53. The antiproliferative effect was identified to be triggered by the induction of cell apoptosis (apoptotic ratio of 26.17%) and cell cycle arrest at S phase. Additionally, AP-PAMAM/p53 transfection has been found to suppress the cell migration and invasion of cancer cells. Finally, the enhanced p53 expression level could be detected after p53 transfection at mRNA and protein levels. Conclusion The PAMAM derivative-mediated p53 delivery could be a promising strategy for achieving tumor gene therapy.