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Systemic genetic transfer of p21WAF1 and GM-CSF utilizing of a novel oligopeptide-based EGF receptor targeting polyplex


Based on the fact that aberrant overexpression of some growth factor receptors was observed in a variety of human cancer cells, a novel nonviral gene delivery system GE7, which contains a 16-amino-acid ligand for identifying EGF receptor was constructed for tumor-targeted gene therapy. Intravenous administration of GE7 system revealed that it has the ability to target β-galactosidase (β-gal) reporter gene into murine hepatoma (Hepa) cells. Owing to the limited antitumor effects elicited by a single-gene transfer, recent efforts to treat malignancy using combined gene therapy have been accomplished with varying degrees of success. In this study, the human cyclin-dependent kinase inhibitor gene p21WAF1 and the murine cytokine gene granulocyte–macrophage colony-stimulating factor (GM-CSF) were used simultaneously for in vivo gene therapy through systemic injection of the EGF R targeted GE7/DNA complex into murine hepatoma-bearing mice. The results demonstrated that combined administration of p21WAF1 and GM-CSF could remarkably inhibit the growth of subcutaneously transplanted hepatoma Hepa cells, and significantly increase the survival rate of tumor-bearing mice. The activities of natural killer (NK) cells and specific cytotoxic T lymphocytes (CTL) were clearly enhanced after combined gene therapy. In vitro experiments showed that p21WAF1 gene transfer exhibited a suppressive function on the growth of Hepa cells and the expression of H-2Kb and B7-1 molecules on Hepa cells increased significantly after combined genes delivery. All these results suggested that the GE7 system was able to target therapeutic genes efficiently to cancer cells, which showed high EGF R expression. The cotransfer of p21WAF1 and GM-CSF genes apparently inhibited the growth of tumors through (a) the arrest of tumor cell growth and (b) the enhancement of systemic antitumor immunity.

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epidermal growth factor receptor


granulocyte–macrophage colony-stimulating factor


cyclin-dependent kinase inhibitors


natural killer


specific cytotoxic T lymphocyte






normal saline


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We thank Mr Ming Yao for his help on animal manipulation, Dr Jingjun Li and Ruijiao Zhao for the immunohistochemistry analysis, Min Bai and Hongmei Song (Department of Immunology, Second Military Medical University, Shanghai, China) for their excellent technical assistance in animal experiments and Mr Zhu Tengfang (Department of Pathology, Shanghai Medical University, Shanghai, China) for his assistance with frozen section preparation. We are also grateful to Professor Jianyun Dong (Department of Immunology, Medical University of South Carolina, SC) and Dr William M Shannon (Senior Vice President, GenPhar Inc., SC) for critical reading of the manuscript and helpful suggestion and comments. This work was supported by grants from the Biotechnology Project, National High Technology Program (Project No. Z-20-01-01).

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Correspondence to Jian-Ren Gu.

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Liu, X., Tian, PK., Ju, DW. et al. Systemic genetic transfer of p21WAF1 and GM-CSF utilizing of a novel oligopeptide-based EGF receptor targeting polyplex. Cancer Gene Ther 10, 529–539 (2003).

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