Gene Therapy Of Hepatocellular Carcinoma Using Plk1 Gene Silencing Mediated By PEGylated MoS₂ Nano-sheets

Objective To study the PEGylated Mo S2 Nano-sheets serving as gene carrier for gene delivery and si Plk1 mediated in vitro gene therapy for hepatocellular carcinoma.Methods 1) Mo S2 nanosheets were synthesized by the Morrison method and then subsequently modified by lipoic acid-modified polyethylene glycol(LA-PEG) and branched polyethylenimine(PEI) with molecular weight(MW) of 25 k Da to obtain Mo S2-PEG-PEI whose exosyndromes were evaluated. 2) The cytotoxicity of Mo S2, Mo S2-PEG, and Mo S2-PEG-PEI was evaluated by MTT assay using different cell lines. 3) The ability of Mo S2-PEG-PEI combin of si RNA was evaluated by agarose gel electrophoresis assay. 4)Human hepatoma cell line(Hep G2 cells) were transfected by si Plk1 which is carried by Mo S2-PEG-PEI.5) After the Hep G2 cells was transfected by si Plk1 which is carried by Mo S2-PEG-PEI,the expression levels of protein and m RNA were evaluated by Western blotting and RT-PCR. Apoptosis of liver cancer cells was evaluated by flow cytometry analysisResults 1) Mo S2-PEG-PEI were modified by LA-PEG and PEI which have a good stability and a positively charged. 2) Mo S2, Mo S2-PEG and Mo S2-PEG-PEI have no toxicity. 3)When the N / P ratio of 5 or more, gel electrophoresis showed Mo S2-PEG-PEI si RNA significantly block. 4) Confocal laser fluorescence microscope revealed Mo S2-PEG-PEI / si RNA successfully transfected Hep G2 cells.5) Western blotting and RT-q PCR results show that, Mo S2-PEG-PEI / si RNA silencing protein expression in Hep G2 cells associated.6) Detected by flow cytometry analysis showed that the Mo S2-PEG-PEI as si Plk1 vector transfected hepatoma cells, can make the hepatoma cells apoptosis.Conclusion Mo S2-PEG-PEI has good biocompatibility, no obvious toxicity. Mo S2-PEG-PEI can be used as a carrier si Plk1 which transfect hepatoma cells with high transfection efficiency and decrease the expression of Plk1 protein in the hepatoma cells, leading to apoptosis of hepatoma cells.Therefore,Mo S2-PEG-PEI can be used as an emerging nanocarriers for gene therap.