Effects of silencing β-catenin gene by RNAi on cell viability of human neuroblastom cell

Abstract

Objective To investigate the inhibition effect of silencing β-catenin gene by RNAi on the biological behavior of BE2C cells in vitro.Methods The control vector PgLV-NC and recombinant lentiviral vector PgLV-R in plasmids were transfected into neuroblastorna BE2C cell line.Stable BE2C cell clones were selected by selection medium containing puromycin.The effect of knockdown of β-catenin on cell proliferation was investigated by CCK8 assay and colony formation.The cell cycle distribution was investigated by flow cytometry.Apoptosis was measured by nuclear staining and flow cytometry.Results The stable inhibition of β-catenin gene expression in human neuroblastoma cell lines BE2C was successfully constructed.Compared with negative control and blank groups,BE2C cell proliferation was inhibited including growth reduction,cell cycle arrest in G0/G1 phase and induction of apoptosis.In cell migration and cell invasion assay,compared with the blank (94.3 ± 9.3,54.3 ± 5.6) and negative control group (62.8 ± 9.8,50.3 ± 5.3),the interference number of migrating cells (29.8 ± 3.8) and transmembrane number (23.5 ± 4.6) were significantly reduced (P＜0.01).Conclusions Blocking the expression of β-catenin gene can inhibit cell proliferation and decrease the cell invasiveness in BE2C cell line.G0/G1 phase arrest and induction of apoptosis could he the underlying mechanism.β-catenin gene could be a potential therapeutic target to treat neuroblastoma. Key words: Neuroblastoma; RNA interference; Protein structure, secondary