[SP-TAT-Apoptin induces G1 arrest in HepG2 cells].

AIM

To investigate the effect of SP-TAT-Apoptin in inducing HepG2 cells apoptosis and the possible application on hepatocellular carcinoma gene therapy.

METHODS

Recombinant gene SP-TAT-Apoptin was amplified by PCR and cloned into the eukaryotic vector plenti6-V5-D-TOPO. After the recombinant plasmid was identified by restriction enzyme digestion analysis and DNA sequencing, CHO cells were stably transfected with SP-TAT-Apoptin gene and the culture supernatant was collected. Then the expression of the fusion protein was detected by RT-PCR and Western blot. HepG2 cells were co-cultured with the supernatant. At various times post co-culture, HepG2 cells were detected by FCM.

RESULTS

The secretory Tat-Apoptin has an additive bystander effect as an anti-cancer therapy in vitro. The recombinant Apoptin was able to be secreted from transfected cells and re-enter adjacent un-transfected HepG2 cells, it can induce HepG2 cells apoptosis and induce G0/G1 arrest.

CONCLUSION

SP-TAT-Apoptin can induce HepG2 cell apoptosis and cell cycle G1 arrest.