Oncolytic adenovirus-mediated shRNA against Apollon inhibits tumor cell growth and enhances antitumor effect of 5-fluorouracil
Oncolytic adenovirus-mediated shRNA against Apollon inhibits tumor cell growth and enhances antitumor effect of 5-fluorouracil
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DOI：
10.1038/gt.2008.6
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发表时间：
2008-04-01
期刊：
影响因子：
5.1
通讯作者：
Liu, X.
中科院分区：
文献类型：
--
作者：
Chu, L.;Gu, J.;Liu, X.
Apollon是一种凋亡蛋白的膜相关抑制剂，可保护细胞免受凋亡，并在某些肿瘤细胞中上调。本研究通过RNA干扰技术敲低Apollon蛋白，观察其对人HeLa、HT-1080和MCF-7细胞生长的影响。构建了含有RNA聚合酶III依赖性U6启动子以表达针对Apollon的短发夹RNA（shRNA）的溶瘤腺病毒（ZD 55-siApollon）。我们的数据表明，ZD 55-siApollon成功地发挥了基因敲低效应，并导致在培养物和无胸腺小鼠体内肿瘤细胞生长的抑制。细胞周期分析、4 '，6-二脒基-2-苯基吲哚染色和蛋白质印迹分析表明，ZD 55-siApollon介导的Apollon抑制诱导凋亡。瘤内注射ZD 55-siApollon显著抑制HT-1080异种移植小鼠中的肿瘤生长。此外，ZD 55-siApollon增强了化疗剂5-氟尿嘧啶的抗肿瘤作用。总之，这些结果表明，通过溶瘤腺病毒-shRNA递送系统去除Apollon为癌症治疗提供了一种有希望的方法。
Apollon, a membrane-associated inhibitor of apoptosis protein, protects cells against apoptosis and is upregulated in certain tumor cells. In this study, the effects of Apollon protein knockdown by RNA interference on the growth of human HeLa, HT-1080 and MCF-7 cells in vitro and in vivo were investigated. An oncolytic adenovirus (ZD55) containing the RNA polymerase III-dependent U6 promoter to express short hairpin RNA (shRNA) directed against Apollon (ZD55-siApollon) was constructed. Our data show that ZD55-siApollon successfully exerts a gene knockdown effect and causes the inhibition of tumor cell growth both in culture and in athymic mice in vivo. Cell cycle analysis, 4',6-diamidino-2-phenylindole staining and western blot analysis reveal that ZD55-siApollon-mediated suppression of Apollon induces apoptosis. Intratumoral injection of ZD55-siApollon significantly inhibits tumor growth in HT-1080 xenograft mice. Furthermore, ZD55-siApollon enhances the antitumor effect of 5-fluorouracil, a chemotherapeutic agent. In conclusion, these results suggest that the depletion of Apollon by oncolytic adenovirus-shRNA delivery system provides a promising method for cancer therapy.