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INHIBITION OF BETA‐CATENIN IN HEPATOCELLULAR CARCINOMA VIA ANTISENSE EFFECTS OF A NOVEL CELL PERMEABLE GUANINE BASED‐PEPTIDE NUCLEIC ACID
Author(s) -
Delgado Evan,
Bahal Raman,
Ly Danith,
Monga Satdarshan Paul
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.115.9
Subject(s) - transfection , viability assay , wnt signaling pathway , microbiology and biotechnology , cancer research , cell growth , cell , chemistry , biology , western blot , signal transduction , biochemistry , gene
Hepatocellular carcinoma (HCC) is the third leading cause of death by cancer worldwide. Lack of efficacious treatment remains a major problem in managing patients diagnosed with HCC. The Wnt/β‐catenin signaling pathway is implicated in around 30% of all cases of HCC specifically due to stabilizing mutations in the β‐catenin gene. Here we report the effect of novel, cell permeable, guanine‐based peptide nucleic acid antisense oligonucleotides designed against either the transcription start site (T1‐AS) or the translation start site (T2‐AS) of the human β‐catenin gene. Transfection efficiency was monitored by a fluorescence tag on the AS. The effect on Wnt signaling was measured by TOPflash luciferase reporter assay, for cell viability by MTT assay, and for molecular mechanism by Western blot analysis on HCC cell extracts after treatment. Both T1 and T2 transfected comparably in HCC cells without aid from transfection reagents demonstrating their high cell permeability. T1 and T2 inhibited β‐catenin activity in HepG2 cells that harbor mutant β‐catenin. A concomitant decrease in HCC cell viability was evident following AS treatment. Western blot analysis identified a significant decrease in total β‐catenin protein in AS treated cells along with decrease in levels of known β‐catenin targets. Thus we have characterized potent AS oligonucletides that robustly target the β‐catenin gene and are highly cell permeable.