Chu L, Gu J, Sun L, Qian Q, Qian C, Liu X
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, PR China.
Gene Ther. 2008 Apr;15(7):484-94. doi: 10.1038/gt.2008.6. Epub 2008 Jan 31.
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.
Apollon是一种膜相关凋亡抑制蛋白，可保护细胞免受凋亡，并在某些肿瘤细胞中上调。在本研究中，研究了通过RNA干扰敲低Apollon蛋白对人HeLa、HT-1080和MCF-7细胞体外和体内生长的影响。构建了一种溶瘤腺病毒（ZD55），其含有RNA聚合酶III依赖性U6启动子，以表达针对Apollon的短发夹RNA（shRNA）（ZD55-siApollon）。我们的数据表明，ZD55-siApollon成功发挥了基因敲低作用，并在体外培养和体内无胸腺小鼠中均导致肿瘤细胞生长受到抑制。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，ZD55-siApollon介导的Apollon抑制可诱导凋亡。瘤内注射ZD55-siApollon可显著抑制HT-1080异种移植小鼠的肿瘤生长。此外，ZD55-siApollon增强了化疗药物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递送系统耗尽Apollon为癌症治疗提供了一种有前景的方法。
