组蛋白与材料协同效应-An archaeal histone-like protein.pdfVIP

组蛋白与材料协同效应-An archaeal histone-like protein.pdf

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Cancer Gene Therapy (2007) 14, 968–975 r 2007 Nature Publishing Group All rights reserved 0929-1903/07 $30.00 /cgt ORIGINAL ARTICLE An archaeal histone-like protein mediates efficient p53 gene transfer and facilitates its anti-cancer effect in vitro and in vivo 1 1 2 2 2 1 1 1 YY Li , R Wang , GL Zhang , YJ Zheng , P Zhu , ZM Zhang , XX Fang and Y Feng 1Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun, China and 2Biotechnology Application Laboratory, Institute of Military Veterinary, Academy of Military Medical Science, Changchun, China 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 b-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 30 -[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

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