Successful gene therapy requires safe and efficient gene vectors and gene delivery methods. This study is to investigate the effects of double suicide genes on the proliferation and apoptosis of HeLa cells by using the ultrasound-targeted microbubble destruction (UTMD). A lentiviral vector with the KDR promoter was constructed, packaged, and delivered into HeLa cells by UTMD. The results showed that the encapsulation efficiency was 90.6 ± 3.1% and the drug-loading efficiency was 29.2 ± 0.9% assessed by reversed-phase high performance liquid chromatography (RP-HPLC). Cell proliferation was determined by MTT assay and apoptosis was detected by flow cytometry. The proliferation rates of HeLa cells were significantly inhibited when treated with dual-gene lentiviral vectors or lentiviral vector-loaded microbubbles plus UTMD (P < 0.05). Moreover, the inhibiting effects were enhanced along with the increased ultrasonic intensities and declined at 24 h post-irradiation. Additionally, in comparison with the control group, the apoptotic rates of HeLa cells were significantly elevated in the lentiviral vector group and the lentiviral vector microbubble groups (P < 0.05). The apoptotic rates were also elevated as the ultrasonic irradiation intensities were increased (P < 0.05). The results suggest that dual-gene lentiviral vector-loaded microbubbles inhibit proliferation and enhance apoptosis of cervical cancer cells.