A Single Recombinant Adenovirus Expressing p53 and p21-targeting Artificial microRNAs Efficiently Induces Apoptosis in Human Cancer Cells
A Single Recombinant Adenovirus Expressing p53 and p21-targeting Artificial microRNAs Efficiently Induces Apoptosis in Human Cancer Cells
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DOI：
10.1158/1078-0432.ccr-08-2396
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发表时间：
2009-06-01
影响因子：
11.5
通讯作者：
Tokino, Takashi
中科院分区：
文献类型：
--
作者：
Idogawa, Masashi;Sasaki, Yasushi;Tokino, Takashi
目的：涉及p53的基因转移被认为是一种潜在有效的癌症治疗方法，但并不能在所有人类癌症中产生良好的治疗反应。p53的激活诱导细胞周期停滞或凋亡。细胞周期阻滞响应p53激活主要是通过诱导细胞周期蛋白依赖性激酶抑制剂p21介导的。由于p21对p53介导的细胞凋亡也具有抑制作用，因此预期p53诱导的p21表达的抑制将导致细胞凋亡的优先诱导。然而，p21也具有肿瘤抑制特性。在本研究中，我们开发了一种同时表达p53和抑制p21的腺病毒载体，以增强p53介导的凋亡。（Ad-p53/miR-p21），其能够使p53蛋白和靶向p21的人工microRNA的共顺反子表达，并检查该载体在体外和体内的治疗效果。p21的表达水平在Ad-p53/miR-p21感染后显著减弱。在结直肠癌和肝细胞癌细胞中，与单独表达p53的腺病毒（Ad-p53/miR-对照）相比，Ad-p53/miR-p21感染增加了细胞凋亡。Ad-p53/miR-p21还显著增加癌细胞对阿霉素（多柔比星）的化学敏感性。在裸小鼠异种移植瘤模型中，与注射Ad-p53/miR-p21对照相比，直接注射Ad-p53/miR-p21后肿瘤体积明显缩小。结论：这些结果表明腺病毒介导的p53和p21特异性microRNA的转导可能有助于人类癌症的基因治疗。
Purpose: Gene transfer involving p53 is viewed as a potentially effective cancer therapy, but does not result in a good therapeutic response in all human cancers. The activation of p53 induces either cell cycle arrest or apoptosis. Cell cycle arrest in response to p53 activation is mediated primarily through the induction of the cyclin-dependent kinase inhibitor p21. Because p21 also has an inhibitory effect on p53-mediated apoptosis, the suppression of p53-induced p21 expression would be expected to result in the preferential induction of apoptosis. However, p21 also has tumor-suppressive properties. In this study, we developed an adenovirus vector that expresses p53 and suppresses p21 simultaneously to enhance p53-mediated apoptosis.Experimental Design: We constructed a replication-deficient recombinant adenovirus (Ad-p53/miR-p21) that enabled cocistronic expression of the p53 protein and artificial microRNAs that targeted p21, and examined the therapeutic effectiveness of this vector in vitro and in vivo.Results: The levels of p21 were significantly attenuated following infection with Ad-p53/miR-p21. In colorectal and hepatocellular carcinoma cells, infection with Ad-p53/miR-p21 augmented apoptosis as compared with an adenovirus that expressed p53 alone (Ad-p53/miR-control). Ad-p53/miR-p21 also significantly increased the chemosensitivity of cancer cells to adriamycin (doxorubicin). In a xenograft tumor model in nude mice, tumor volume was significantly decreased following the direct injection of Ad-p53/miR-p21 into the tumor, as compared with the injection of Ad-p53/miR-control.Conclusion: These results suggest that adenovirus-mediated transduction of p53 and p21-specific microRNAs may be useful for gene therapy of human cancers.