Objective: To construct recombinant plasmid CD147-shRNA and detect its inhibitory effect on CD147 expression of hepatoma cell line SMMC- 7721 and HepG2.
Methods: Three characteristic target sequences were designed according to the sequence of CD147 in Genebank,and the synthesized polynucleotide was connected to pBS/U6.The recombinant plasmid was identified by restriction enzyme digestion and sequencing and transfected into hepatoma cells SMMC-7721 and HepG2. The expression of CD147 was detected by Western blot and immunofluorescence.
Results: CD147-shRNAs recombinant plasmid was completely and correctly constructed comfirmed by restriction enzyme digestion and sequencing. All the plasmids had gene silencing effect on CD147 expression of HepG2, but the inhibit effect of pBS/U6/CD147- shRNA3 was most significant.
Conclusions: We succesfully constructed pBS/U6/CD147-shRNA and screened out pBS/U6/CD147-shRNA3 which has the best gene silence effect.