Objective: To investigate the possibility of using generation 5 polyamidoamine dendrimers (G5-PAMAM-D) as gene vector for eukaryotic expression plasmid of siRNA in prostate carcinoma in vitro and vivo.
Methods: Firstly, eukaryotic expression vector of siRNA pSilencing 4.1-EGFP-shRNA, specific for enhanced green fluorescent protein (EGFP), pSilencing 4.1-STAT3-shRNA for signal transducers and activators of transcription 3 (STAT3) was constructed. pEGFP-C1 and pSilencing 4.1-EGFP-shRNA were cotransfected into prostate cancer cells PC-3 and 22Rv1 with G5-PAPAM-D as vector, and to observe silencing of EGFP. Next, pSilencing 4.1-STAT3-shRNA was transfected into PC-3 and 22Rv1 cells by G5-PAPAM-D, Western blotting and apoptosis staining was used to detect silencing of STAT3 and growth inhibition. Thirdly, BALB/C mice subcutaneous tumor model was made with PC-3 cells. Polyplex of G5-PAMAM-D and pSilencing 4.1-STAT3-shRNA was injected intratumorally. The tumor volume was measured and recorded.
Results: Fluorescence detection and Western blotting analysis demonstrated that G5-PAMAM-D was able to deliver Silencing 4.1-EGFP-shRNA and pSilencing 4.1-STAT3-shRNA into the two prostate cancer cell lines, and shRNA was expressed to induce silence of EGFP and STAT3. MTT results showed that proliferation of prostate cancer cells was suppressed by G5-PAMAM-D/pSilencing 4.1-STAT3-shRNA and induced apoptosis of PC-3 cells in vitro. Human prostate cancer in mice was successfully formed by inoculation of PC-3 cells into male BABL/C mice. In G5-PAMAM-D/pSilencing 4.1-STAT3-shRNA treated group, the tumor volume was shrank remarkably at 9 days after treatment and tumor growth was retarded compared with control groups.
Conclusion: GS-PAMAM-D nanoparticles can be used to deliver plasmid vector expressing shRNA into prostate cancer cells effectively in vitro and vivo. It appears to be a promising gene vector for RNA interference therapy in prostate cancer.