In vitro RNA interference against beta-catenin inhibiting the proliferation of pediatric hepatic tumors

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To address beta-catenin's capacity in maintaining the malignant phenotype in established pediatric hepatoblastoma and hepatocellular carcinoma cell lines, HuH-6,and HepG2,harboring mutated and overexpressed beta-catenin, we carried out a series of in vitro analyses through a transfection of short interfering RNAs to generate a loss of function model. HuH-7,another hepatoblastoma cell line derived from a pediatric patient without a stabilizing mutation was used for comparison. RNA interference successfully manipulated the degradation of overexpressed beta-catenin. In all cell lines, beta-catenin mRNA was suppressed by 80-90 % after 48 h of transfection, and a reduction of its protein expression was demonstrated. In HuH-6 and HepG2,the pre-existing beta-catenin nuclear accumulation disappeared and reductions of beta-catenin downstream target genes, c-myc and cyclinD1,were also evidenced after treatment. The in vitro proliferation of both cell lines was transiently inhibited. In contrast, the suppression of beta-catenin in HuH-7 did not lead to a significant change in the expression of target genes or cellular proliferation. Our data indicate that beta-catenin can be considered a specific target for gene therapy in pediatric hepatic tumors with mutations and overexpression of this gene.