The experimental study of two transfection strategies of recombinant plasmid into Tca8113 cells

Objective:To investigate the transfection efficiency and toxicity of novel cationic polymer cross-linked polyethylenimine-ethyleneglycol dimethacrylate(PEI-EGDMA) and cationic liposome TRANSfection. Methods:PEI-EGDMA and TRANSfection were used to deliver PRNAT-U6.1/Neo vector containing VEGF shRNA to Tca8113 cells.GFP gene was inserted into the vector as a report gene. Four kinds of concentration gradient were set up.The cells were observed by fluorescent microscope 24 h after transfection. The fluorescence positive cells were counted and transfection efficiency were calculated. Evaluation of cytotoxicity was performed by MTT assay 48 h after transfection. Results:The best transfection efficiency could be got when the ratio of PEI-EGDMA∶plasmid and TRANSfection∶plasmid(W∶W) is 1.5∶1 and 2∶1, around 72% and 55% respectively. It had significant difference(P0.05)compared with other concentration gradient groups. Furthermore, there was significant difference in their optimal transfection condition, the effective rate of transfection of PEI-EGDMA was increased significantly. MTT results indicated no cell growth difference between experimental groups and blank control group.Conclusion:PEI-EGDMA could mediate more efficient transfection compared with liposome TRANSfection.It also has low cell toxicity. It could be a ideal vector in gene transfection therapy treating human tongue squamous cell carcinoma.