Abstract:AIM: To observe human lens epithelial cells(HLE-B3)proliferation changes when the PAX6 gene has been knocked-down.
METHODS: Four groups PAX6 shRNA lentiviral vectors and the control group lentiviral(pGCL-GFP-shRP1, 2, 3, 4, NC)infect B3 cells. Real Time PCR and Western blot analysis was used to determine PAX6 silence after infection 96 hours. Effective PAX6-silence pGCL-GFP-shRP4 was selected to infect B3 cells, MOI(Multiplicity of infection)= 10 and the changes were observed in the number of the cells. The B3 cells apoptosis was detect by flow cytometry after 48 hours in infection.
RESULTS: RNAi-virus-infected B3 cells decreased in the number of cells with infection prolonged. The flow cytometry results showed that after knock-down the PAX6 for 48 hours, apoptotic peak appeared before the G1/G0 phase.
CONCLUSION: Lentivirus-mediated RNA interference can effectively silence PAX6 expression in B3 cells. There is HLE-B3 apoptosis after PAX6 gene knock-down.
