RRC ID 72284
Author Sangkhathat S, Kusafuka T, Miao J, Yoneda A, Nara K, Yamamoto S, Kaneda Y, Fukuzawa M.
Title In vitro RNA interference against beta-catenin inhibits the proliferation of pediatric hepatic tumors.
Journal Int J Oncol
Abstract Mutations of beta-catenin have been identified in the majority of pediatric hepatic malignancies, including hepatoblastoma (HB) and hepatocellular carcinoma (HCC), suggesting its important contribution in hepatic tumorigenesis in this age group. However, the role of beta-catenin/canonical Wnt signaling pathway in the neoplastic growth of cancer cells has not been directly studied. To address beta-catenin's capability in maintaining the malignant phenotype in established pediatric HB and HCC 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 (siRNAs) to generate a loss-of-function model. HuH-7, another HB 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 the 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.
Volume 28(3)
Pages 715-22
Published 2006-3-1
PMID 16465377
MeSH Apoptosis Blotting, Western Cell Line, Tumor Cell Nucleus / metabolism Cell Proliferation* Child DNA Mutational Analysis Gene Expression Regulation, Neoplastic Humans Liver Neoplasms / genetics Liver Neoplasms / pathology Liver Neoplasms / physiopathology Mutation RNA Interference* RNA, Small Interfering / genetics Transfection beta Catenin / genetics* beta Catenin / metabolism
Resource
Human and Animal Cells Hep G2 HuH-7