Inhibition of cell proliferation and migration through nucleobase-modified polyamidoamine-mediated p53 delivery

• Published in: International journal of nanomedicine 2018-01, Vol.13, p.1297-1311
• Main Authors: Han, Haobo, Chen, Wenqi, Yang, Jiebing, Liang, Xiao, Wang, Yudi, Li, Quanshun, Yang, Yan, Li, Kun
• Format: Article
• Language: English
• Subjects: Amino acids; Apoptosis - drug effects; Biomedical materials; Cancer; Cancer cells; Cancer therapies; Carcinoma; Cell cycle; Cell Cycle - drug effects; Cell growth; Cell Line, Tumor; Cell Movement - drug effects; Cell Proliferation - drug effects; Cervical cancer; Chondroitin sulfate; Cytotoxicity; Dendrimers - chemical synthesis; Dendrimers - chemistry; Deoxyribonucleic acid; DNA; EDTA; Efficiency; Female; Gene therapy; Genes; HeLa Cells; Humans; Infection; Laboratories; Lung cancer; Membrane Potential, Mitochondrial - drug effects; Messenger RNA; Nanoparticles; Nanoparticles - chemistry; Original Research; Plasmids; Polyamines - chemistry; Polymerase chain reaction; RNA; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transfection; Tumor proteins; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Tumors; Wound care; Wound healing; Wound Healing - drug effects; Wounds
• ISSN: 1178-2013; 1176-9114; 1178-2013
• DOI: 10.2147/IJN.S146917

The nucleobase 2-amino-6-chloropurine-modified polyamidoamine (AP-PAMAM) was used as a carrier for gene delivery to achieve the antitumor effects. The condensation of 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. The carrier could condense 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 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 transfection at mRNA and protein levels. The PAMAM derivative-mediated delivery could be a promising strategy for achieving tumor gene therapy.