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In vivo bioluminescent imaging of α‐fetoprotein‐producing hepatocellular carcinoma in the diethylnitrosamine‐treated mouse using recombinant adenoviral vector
Author(s) -
Kim Kwang Il,
Park Ju Hui,
Lee Yong Jin,
Lee Tae Sup,
Park Jae Jun,
Song Inho,
Nahm SangSoep,
Cheon Gi Jeong,
Lim Sang Moo,
Chung JuneKey,
Kang Joo Hyun
Publication year - 2012
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.2648
Subject(s) - luciferase , in vivo , reporter gene , genetic enhancement , bioluminescence imaging , microbiology and biotechnology , viral vector , recombinant dna , hepatocellular carcinoma , carcinogenesis , gene expression , cancer research , biology , transgene , gene , transfection , biochemistry
Background The in vivo molecular imaging method is a useful tool for monitoring carcinogenesis in various hepatocellular carcinoma (HCC) models, such as xenografted‐, chemical induced‐ and transgenic mice. The tumor‐specific gene expression strategy, such as transcriptional targeting, is essential for achieving a lower toxicity for normal liver tissue in therapy and the monitoring of tumor progression in diagnosis, respectively. The present study aimed to visualize spontaneously developing α‐fetoprotein (AFP)‐producing HCC through targeted gene expression in tumors using recombinant adenoviral vector. Methods The recombinant adenovirus vector, AdAFPfLuc (containing firefly luciferase gene driven by human AFP enhancer/promoter) was prepared. After in vitro infection by adenovirus, gene expression was confirmed using the luciferase assay, semi‐quantitative reverse transcriptase‐polymerase chain reaction and western blotting in AFP‐producing and nonproducing cells. Tumor‐bearing mice were intravenously injected with adenovirus, and bioluminescent images were obtained. Results The expression of fLuc was efficiently demonstrated by the luciferase assay in AFP‐producing cells but not in AFP‐nonproducing cells. AFP‐producing HCC targeted gene expression was confirmed at the mRNA and protein levels. After being injected intravenously in HuH‐7 xenografts and HCC‐bearing diethylnitrosamine‐treated mice using adenovirus, functional reporter gene expression was confirmed in tumors by in vivo bioluminescent imaging (BLI). Conclusions The recombinant adenovirus vector system can be used to monitor spontaneously developing AFP‐producing HCC and to evaluate targeted gene expression in tumors by in vivo BLI in a small animal model. Copyright © 2012 John Wiley & Sons, Ltd.