BACKGROUND: The introduction of exogenous Rb gene into retinoblastoma is an effective way to inhibit growth of the tumor. piggyBac transposon as a vector is safe and highly efficient, which is a potential vector on gene therapy.
OBJECTIVE: To explore the transfection efficiency and its effect on inhibiting retinoblastoma by transfecting human retinoblastoma cell line SO-RB50 with an exogenous Rb gene expression system mediated by piggyBac transposon.
METHODS: pPiggyBac-Rb plasmid that contains a piggyBac transposon sequence and express normal RB gene was constructed and used to transfect SO-RB50 cells alone or with pPiggyBac-helper. Coomassie brilliant blue staining, quantitative real-time PCR and immunofluorescence were adopted to evaluate the transfect efficiency of exogenous Rb gene mediated by piggyBac transposon. The viability of retinoblastoma cells was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
RESULTS AND CONCLUSION: Transfection efficiency was the highest with help of piggyBac transposon. piggyBac transposon-mediated exogenous Rb gene integrated into genome and obtained a long-term and stable expression．It also had a remarkable inhibition effect on the viability of SO-RB50. All these indicate that piggyBac transposon is a potential vector for retinoblastoma gene therapy.