Overexpression of Survivin mutant Thr34Ala induces apoptosis and inhibits gastric cancer growth.

ABSTRACT

UNLABELLED Survivin is highly expressed in human gastric cancer and correlated with chemoresistance and poor prognosis. In this study, we explored the effect of adeno-associated virus (AAV)-mediated survivin dominant mutant Thr34Ala (rAAV-Sur-Mut(T34A)) on gastric cancer growth. AAV-Sur-Mut (T34A) virus was generated and purified using the AAV vector pAM/CAG-WPRE.poly(A) vector. Cell proliferation was determined by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) Assay. Apoptosis was determined by Flow cytometry and Terminal deoxynucleotidyl transferase dUTP nick end labeling. Gene expression was determined by Western blot and immunohistochemistry. Tumor growth was evaluated using a xenograft mouse model. Overexpression of survivin promoted cell growth of gastric cancer cells. Infection of rAAV-Sur-Mut(T34A) virus inhibited cell proliferation, induced apoptosis and sensitized gastric cancer cells to 5-Fluorouracil in vitro. Treatment of rAAV-Sur-Mut(T34A) significantly inhibited cell proliferation, induced apoptosis and inhibited gastric cancer growth in vivo. Our results suggest that the combination of rAAV-Sur-Mut(T34A) and chemotherapy may be a new approach for gastric cancer therapy. KEYWORDS Survivin, survivin mutant T34A, adeno-associated virus, gastric cancer, apoptosis, gene therapy.