Efficient Gene Transfer to Hepatoblastoma Cells through Asialoglycoprotein Receptor and Expression under the Control of the Cyclin A Promoter.

Aramaki, Yukihiko; Lee, Insugi; Arima, Hisatomi; Sakamoto, Takatoshi; Magami, Yasushi; Yoshimoto, Takayuki; Mizuguchi, Junichiro; Koyanagi, Yasuhisa; Nikaido, Toshio; Tsuchiya, Satoshi

doi:10.1248/bpb.26.357

Cited by 10 publications

(5 citation statements)

References 24 publications

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“…However, only the HepG2 cells were used in these investigations so far [23,24]. The HepG2 cell line was derived from a patient with a hepatocellular carcinoma (HCC) as demonstrated in the original publication and as seen in our own previous studies [7,25].…”

Section: Discussionmentioning

confidence: 99%

Optimizing vector application for gene transfer into human hepatoblastoma cells

et al. 2006

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Gene targeting is currently of distinct interest as an innovative additive treatment option in various malignancies. Its role in pediatric liver tumors has not yet been evaluated thoroughly. For the first time the authors systematically analyzed both lipid-based transfection as well as transduction with adenovirus vectors (Ad) and Sendai virus vectors (SeVV) in order to optimize gene transfer into hepatoblastoma (HB) cells. Two HB cell lines were infected with Ad or SeVV coding for green fluorescent protein (Ad-GFP, SeVV-GFP); transduction efficiencies and apoptosis were assessed using flow cytometry. Furthermore, lipofection of HB cell lines with plasmid-constructs comprising liver-specific promoters was performed using Lipofectamine 2000 and FuGENE 6; lipofection efficiency was monitored by flow cytometry, microscopy, and luciferase activity. The Ad-GFP showed higher transduction rates (61-86%) than the SeVV-GFP (4-24%) depending on the HB cell line used. Infections with first generation SeVV vectors (SeVV-GFP) led to increased target cell apoptosis (7-43%) compared to Ad-GFP (4-16%). The Lipofectamine 2000 revealed a higher transfection efficiency than the FuGENE 6 for both HB cell lines tested. The liver-specific promoters were found to be differently active in the HB cell lines. This study delineates recombinant adenovirus vectors as a promising tool for gene transduction in the HB cells. Furthermore, enhanced activity of the liver-specific promoters in HUH6 cells compared to HepT1 cells supports the observation of varying biological behavior in histologically differing HB tissues.

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Section: Discussionmentioning

confidence: 99%

Optimizing vector application for gene transfer into human hepatoblastoma cells

et al. 2006

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“…Adenoviral vectors are common tools for gene delivery in different tumor models [24][25][26][27][28]. Treatment approaches include viral vectors coding for cytokines, suicide genes, anti-angiogenic factors, and cell cycle modulators.…”

Section: Discussionmentioning

confidence: 99%

In vitro gene targeting in human Hepatoblastoma

et al. 2005

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Poor treatment results in advanced hepatoblastoma (HB) made alternative treatment approaches desirable. Gene-directed tumor therapy is increasingly investigated in different malignancies. The aim of this study was to analyze possible alternatives of gene transfer into HB cells and to study therapeutic applications based on different strategies. Liposomal transfection of HB cells was assessed using liver-specific promoters, and adenovirus and Sendai virus transductions were performed in vitro. Transfer efficiencies were measured via flow cytometry determining expression of vector-encoded marker gene green fluorescent protein. Gene silencing of the anti-apoptotic bcl-2 gene in HUH6 cells was performed using lipofection of small interfering RNA (siRNA). Additionally, suicide gene therapy was carried out through a yeast-derived cytosine deaminase (YCD)-combined yeast uracil phosphoribosyltransferase (YUPRT)-based adenovirus-mediated gene transfer, leading to a potent intracellular prodrug transformation of 5-fluorocytosine into 5-fluorouracil. Treatment efficiencies were monitored via MTT viability assay. Highest gene transfer rates (86%) were observed using adenovirus transduction. We furthermore observed a significant therapeutic effect of adenovirus-mediated YCD::YUPRT suicide gene transfer. Liposomal-mediated anti-bcl-2 siRNA transfer led to a significant improvement of cisplatin treatment in HUH6 cells. Liver-specific promoters were found to be strongly active in HUH6 cells (mixed HB-derived), but less active in HepT1 cells (embryonal HB-derived). Liposomal transfection and viral transduction are effective approaches to genetically manipulate HB cells in vitro. For the first time, we demonstrate a positive effect of siRNA gene silencing in this malignancy. Additionally, we successfully investigated a model of adenovirus-based suicide gene therapy in HB cell cultures. Our data strongly encourage further studies assessing these alternative treatment approaches.

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“…Yet, analyses on pediatric liver tumors are sporadic and mainly include pediatric hepatocellular carcinoma (HCC). [35][36][37] The cells used in these studies (HepG2) were referred to as being of hepatoblastoma origin. However, according to the original publication and various other reports, HepG2 cells originate from a child with HCC [38][39][40].…”

Section: Gene Transfer Systems For Hepatoblastomamentioning

confidence: 99%