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Inhibitory effect of polyamidoamine dendrimer-CXCR4-shRNA nanoparticles on the proliferation of renal carcinoma cells
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Abstract:
Objective: To optimize the preparation of polyamidoamine dendrimer-CXCR4-shRNA nanoparticles (PAMAM-shRNA) and study the inhibitory effect of PAMAM-shRNA nanoparticles on the proliferation of renal carcinoma cells in vitro. Methods: The seventh generation of polyamidoamine dendrimer (PAMAM-D) and CXCR4-shRNA were mixed at a mass ratio of 1∶0.73) at room temperature. The morphology and structure of PAMAM-shRNA by transmission electron microscopy, and determined the size of the nanoparticles were analyzed by laser particle size analyzer. Human renal carcinoma A498 cells were transfected with PAMAM-shRNA,CXCR4-shRNA and PAMAM-D respectively. After transfection, cell viability was assessed by MTT assays, cell apoptosis by flow cytometry, and CXCR4 mRNA abundance by Real-time PCR. Results: The prepared PAMAM-shRNA nanoparticles were evenly distributed and non-adherent with a mean diameter of 176.5±25.48 nm. PAMAM-shRNA effectively inhibited A498 proliferation in time- and dose-dependent manners, and the highest proliferation inhibition rate was up to （66.5±2.7)%. PAMAM-shRNA induced A498 cell apoptosis. CXCR4 mRNA abundance was significantly decreased (P<0.05) in cells transfected with PAMAM-shRNA (0.29±0035) than in cells transfected with CXCR4-shRNA (0.70±0.084). Conclusions: PAMAM dendrimers may efficiently mediate the entry of CXCR4-shRNA into renal carcinoma cells, where they exert proliferation-inhibitory and apoptosis-promoting activities. These observations suggest that PAMAM-shRNA nanoparticles may have a potential clinical application in gene therapy of renal cancer.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No. 81202019），and the Youth Research Project of Shanghai Health Bureau（No. 2011Y197）
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