Dang S C, Feng S, Wang P J, Cui L, Qu J G, Zhang J X
Neoplasma. 2015;62(1):81-7. doi: 10.4149/neo_2015_010.
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.
Survivin, survivin mutant T34A, adeno-associated virus, gastric cancer, apoptosis, gene therapy.
生存素在人类胃癌中高表达，与化疗耐药性和不良预后相关。在本研究中，我们探讨了腺相关病毒（AAV）介导的生存素显性突变体Thr34Ala（rAAV-Sur-Mut(T34A)）对胃癌生长的影响。使用AAV载体pAM/CAG-WPRE.poly(A)载体产生并纯化AAV-Sur-Mut (T34A)病毒。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定细胞增殖。通过流式细胞术和末端脱氧核苷酸转移酶dUTP缺口末端标记法测定细胞凋亡。通过蛋白质免疫印迹法和免疫组织化学法测定基因表达。使用异种移植小鼠模型评估肿瘤生长。生存素的过表达促进胃癌细胞的生长。rAAV-Sur-Mut(T34A)病毒感染在体外抑制细胞增殖、诱导细胞凋亡并使胃癌细胞对5-氟尿嘧啶敏感。rAAV-Sur-Mut(T34A)治疗在体内显著抑制细胞增殖、诱导细胞凋亡并抑制胃癌生长。我们的结果表明，rAAV-Sur-Mut(T34A)与化疗联合可能是一种新的胃癌治疗方法。
生存素；生存素突变体T34A；腺相关病毒；胃癌；细胞凋亡；基因治疗
