An archaeal histone-like protein mediates efficient p53 gene transfer and facilitates its anti-cancer effect in vitro and in vivo
An archaeal histone-like protein mediates efficient p53 gene transfer and facilitates its anti-cancer effect in vitro and in vivo
复制标题
古细菌组蛋白样蛋白介导有效的 p53 基因转移并促进其体外和体内抗癌作用
DOI：
10.1038/sj.cgt.7701086
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发表时间：
2007-12-01
影响因子：
6.4
通讯作者：
Feng, Y.
中科院分区：
文献类型：
--
作者：
Li, Y. Y.;Wang, R.;Feng, Y.
提高非病毒载体基因传递系统的转染效率是其在基因治疗中应用的关键。我们先前描述了一种基于古细菌组蛋白样蛋白（HPhA）的基因递送系统，并表明HPhA与核酸形成稳定的非共价复合物，并通过使用β-半乳糖苷酶作为报告基因来改善其递送。本研究以HPhA为载体，将野生型p53基因转染到肿瘤细胞中，并在体外和体内检测p53基因的表达水平和活性。通过实时逆转录酶-PCR和蛋白质印迹分析确定基因表达。分别通过XTT（3 '-[1-（苯氨基羰基）-3，4-四氮唑]-双（4-甲氧基-6-硝基）苯磺酸钠水合物）测定和膜联蛋白V-FITC（异硫氰酸荧光素）染色评估HPhA介导的p53转染的细胞生长抑制和细胞凋亡。此外，将HPhA/p53转染到CNE（鼻咽癌细胞系）异种移植的裸鼠中，并测量肿瘤生长。本研究表明，HPhA提高了p53基因转移的效率和抗肿瘤活性相比，广泛使用的Lipofectamine。这些结果表明，HPhA增强了p53基因转移的体外和体内效率，并建议它可能作为一个有前途的工具，基因递送和基因治疗。
The improvement of the transfection efficiency of the non-viral-based gene delivery systems is a key issue for the application in gene therapy. We have previously described an archaeal histone-like protein-based (HPhA) gene delivery system and showed that HPhA formed stable non-covalent complexes with nucleic acids and improved their delivery by using β-galactosidase as a reporter gene. In this study, the wild-type p53 gene was transfected into the cancer cells using the HPhA as a vector, and the expression level and the activity of p53 gene were evaluated both in vitro and in vivo. Gene expression was determined by real-time reverse transcriptase-PCR and western blotting analysis. The cellular growth inhibition and apoptosis of HPhA-mediated p53 transfection were assessed by XTT (sodium 3′-[1-(phenylaminocarbonyl)-3, 4-tetrazolium]-bis (4-methoxy-6-nitro) benzene sulfonic acid hydrate) assay and annexin V–FITC (fluorescein isothiocyanate) staining, respectively. Further more, transfection of HPhA/p53 into CNE (nasopharyngeal carcinoma cell line)-xenografted nude mice was performed and tumor growth was measured. The present study demonstrates that HPhA enhances the efficiency of p53 gene transfer and antitumor activity compared with the widely used Lipofectamine. These results demonstrate that HPhA enhances the in vitro and in vivo efficiency of p53 gene transfer and suggest that it may be served as a promising tool for gene delivery and gene therapy.