ShRNA targeting against CTNNB1 inhibits growth of human colon cancer Lovo cells

Objective To observe the inhibitory effect of CTNNB1 gene expression and cell proliferation of the human colon cancer cell line Lovo. Methods The shRNA plasmid vector against CTNNB1 was constructed and transfected into Lovo cells with LipofectamineTM2000. The down-regulations of CTNNB1 expressions were detected by RT-PCR and western blot analysis. The cell proliferation inhibitions were determined by MTT assay. The effect of these shRNAs on cell cycle distribution and apoptosis were examined by flow cytometry. Results The shRNA vectors targeted against CTNNB1 were successfully constructed and efficiently suppressed the expression of CTNNB1 mRNA and protein(P<0.05). The expression inhibition rates were 40.63% and 44.79% at the mRNA and protein level respectively. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates on a time-dependent manner. At 72 hours post-transfection,the cell viability of CTN group was 45.8%,which was statistical significance when compared with that of blank control group (P<0.05). Moreover, the cancer cells showed significant G0/G1 arrest and increased apoptosis at 72 hours after transfection. Conclusions The specific shRNAs targeted against CTNNB1 could down-regulate the expression of CTNNB1 gene, increase apoptosis and inhibit the growth of Lovo cells. Key words: CTNNB1;  Colon cancer;  RNA interference;  shRNA