Abstract:Objective To investigate the effect of wildtype PTEN gene mediated by adenovirus vector on the cell cycle of human breast cancer cell line MDAMB468 in vitro.
Methods PTEN gene was cloned into pAdTrackCMVPTEN. The newly recombinated pAdPTEN containing green fluorescent protein (GFP) was infected into human breast cancer cell line MDAMB468 in vitro, and the cell cycle was detected by flow cytometric analysis.
Results The recombinant adenovirus vector carrying PTEN was constructed successfully, and the titre of purified adenovirus was 2.5×1010 pfu/mL. Cell number of control group in G1 phase accounted for 41.18%, while the transfected PTEN gene group was 56.47% of G1 cell number as measured by flow cytometry.
Conclusions The recombinant adenovirus vector of PTEN was rapidly constructed successfully using AdEasy1 system. Target genes were successfully transfected and expressed stably. PTEN may cause cell cycle arrest at the G1 phase.