An emerging hybrid nanovector integrating the merits of both viral and nonviral vectors has attracted much attention as the next generation of promising gene vectors to overcome the primary challenge of cancer gene therapy. Due to its inherent advantages, lentivirus (Lv) has been increasingly applied and investigated in medical fields. Herein, a new hybrid nanovector (SS-HPT/Lv) composed of the Lv core and reduction-responsive hyperbranched polyaminoglycoside (SS-HPT) shell is designed via electrostatic interaction. In comparison with polybrene (commercial enhanced transfection reagent), SS-HPT endows the hybrid nanovector with better biosafety. Furthermore, both the appropriate nanoparticle sizes and positive surface potentials contribute to the endocytosis and rescue of SS-HPT/Lv from endocytic vesicles. Regarding therapeutic application, lentiviral vector acquires permanent transgene expression with the capability of integrating to the host chromosome, and SS-HPT/Lv exhibits an improved transduction efficacy. The cytosine deaminase/5-fluorocytosine suicide gene therapy system mediated by SS-HPT/Lv performs an enhanced antitumor efficiency and extends the survival time of glioma-bearing rats. Such a hybrid nanovector strategy would open a new avenue to the development of gene vectors for treating malignant cancers.