Enhancement of p53 gene transfer efficiency in hepatic tumor mediated by transferrin receptor through trans-arterial delivery.
Transferrin-DNA complex mediated by transferrin receptor in combination with interventional trans-arterial injection into a target organ may be a duel-target-oriented delivery means to achieve an efficient gene therapy. In this study, transferrin receptor expression in normal human hepatocyte and two hepatocellular-carcinoma cells (Huh7/SK-Hep1) was determined. p53-LipofectAMINE with different amounts of transferrin was transfected into the cells and the gene transfection efficiency was evaluated. After VX2 rabbit hepatocarcinoma model was established, the transferrin-p53-LipofectAMINE complex was delivered into the hepatic artery via interventional techniques to analyze the therapeutic p53 gene transfer efficiency in vivo by Western blot, immunohistochemical/immunofluorescence staining analysis and survival time. The results were transferrin receptor expression in Huh7 and SK-Hep1 cells was higher than in normal hepatocyte. Transfection efficiency of p53 was increased in vitro in both Huh7 and SK-Hep1 cells with increasing transferrin in a dose-dependent manner. As compared to intravenous administration, interventional injection of p53-gene complex into hepatic tumor mediated by transferrin-receptor, could enhance the gene transfer efficiency in vivo as evaluated by Western blot, immunohistochemical/immunofluorenscence staining analyses and improved animal survival (H = 12.567, p = 0.0019). These findings show the transferrin-transferrin receptor system combined with interventional techniques enhanced p53-gene transfer to hepatic tumor and the duel-target-oriented gene delivery may be an effective approach for gene therapy.
Lu
Qin
Q
Department of Radiology, Molecular Image Laboratory, Zhong-da Hospital, Southeast University, Nanjing, China.
Teng
Gao-Jun
GJ
Zhang
Yue
Y
Niu
Huan-Zhang
HZ
Zhu
Guang-Yu
GY
An
Yan-Li
YL
Yu
Hui
H
Li
Guo-Zhao
GZ
Qiu
Ding-Hong
DH
Wu
Chuan-Ging
CG
D000818:Animals; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D019084:Fluorescent Antibody Technique, Indirect; D018014:Gene Transfer Techniques; D016158:Genes, p53; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry; D008081:Liposomes; D008113:Liver Neoplasms / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*; D011859:Radiography; D011990:Receptors, Transferrin / Q000378:metabolism*; D016019:Survival Analysis; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014168:Transferrin / Q000235:genetics; D019076:Transgenes; D047368:Tumor Burden; D023041:Xenograft Model Antitumor Assays / Q000379:methods
D008081:Liposomes; D011990:Receptors, Transferrin; D014168:Transferrin