Effects of small interfering RNA transfection of B cell lymphoma/leukemia-2-associated athanogene-1 gene on biological characteristics of hepatocellular carcinoma cells

Objective To investigate the effect of B cell lymphoma/leukemia-2-associated athanogene-1 gene (BAG-1) gene expression by RNA interference on the proliferation, apoptosis and Wnt/β-catenin signaling pathway in hepatocellular carcinoma cells. Methods Human normal liver cells L02 as control cells, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of BAG-1 in human hepatocellular carcinoma cell lines HepG2, Huh7, MHCC97L, MHCC97H and HCCLM3; the negative small interfering RNA (siRNA) and BAG-1-siRNA transfected HCCLM3 cells, and not transfected cells for the blank control group, transfection effect was detected after cells transfected for 48 h; the proliferation of the cells was detected by cell counting kit-8 (CCK-8) after cells were transfected at 24, 48 and 72 h; the cells apoptosis after cells were transfected for 48 h was detected by flow cytometry; β-catenin, Survivin, c-Myc protein expression were detected by Western blotting. Results The expression of BAG-1 in hepatocellular carcinoma cells were significantly higher than those of control cells L02 ((mRNA: PHepG2=0.000, PHuh7=0.000, PMHCC97L=0.000, PMHCC97H=0.000, PHCCLM3=0.000; protein: PHepG2=0.030, PHuh7=0.001, PMHCC97L=0.000, PMHCC97H=0.000, PHCCLM3=0.000); the expression of BAG-1 protein (0.097±0.010) after BAG-1-siRNA were transfected into HCCLM3 cells was significantly lower than the control Group (0.489±0.039) (P=0.000); the cells proliferation after BAG-1-siRNA transfected HCCLM3 cells for 48 h (0.312±0.024) and 72 h (0.673±0.051)were significantly lower than the control group (0.448±0.032), (0.673±0.051) (P48 h=0.004, P72 h=0.002); compared with the control group (2.13±0.11)%, (0.512±0.048), (0.349±0.032), (0.166±0.015), the apoptosis rate (13.47±1.05)% in BAG-1-siRNA group increased significantly, β-catenin (0.247±0.028), Survivin (0.153±0.017), c-Myc (0.089±0.009), protein expression was down regulated (Pβ-catenin=0.001, PSurvivin=0.001, Pc-Myc=0.002). Conclusion Inhibiting of BAG-1 expression can reduce the proliferation of hepatoma cells and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway. Key words: B cell lymphoma/leukemia-2-associated athanogene-1 gene; Liver cancer; Proliferation; Apoptosis; Wnt/β-catenin signaling pathway