NF-κB silencing combined with matrine-induced apoptosis in hepatocellular carcinoma cells and expression of related molecules / 第二军医大学学报

ABSTRACT

Objective: To investigate the apoptosis in hepatocellular carcinoma cells induced by NF-κB silencing combined with matrine (MT) and the expression of related molecules. Methods: The recombinant eukaaryotic expression plasmid NF-κB/ P65 siRNA was constructed and transfected into HepG2 cells. RT-PCR was used to detect the efficiency of the constructed RNAi in silencing NF-κB/P65, and the steadily transfected clones of NF-κB/P65 RNAi were selected. The cultured HepG2 cells was randomly divided into four groups control group, MT group (1.5 g/L), steadily transfected group and combination group(steadily transfected cells+MT). The apoptosis of carcinoma cells was analyzed by flow cytometry; expression of NF-κB/P65, CD95(Fas), Smac, and Survivin mRNA and protein in carcinoma cells was examined by RT-PCR and Western blotting analysis. Results: The expression of NF-κB/P65, CD95(Fas), Smac, and Survivin mRNA and protein in MT group was significantly increased compared with that in the control group (P<0.05). The expression of CD95 and Smac in the steadily transfected group was significantly higher than that in the control group (P<0.05), and the expression of NF-κB/P65 and Survivin was significantly suppressed compared with the control group and the MT group(P<0.05). The expression of CD95 and Smac in the combination group was significantly increased compared with that in the other three groups(P<0. 05), and the expression of NF-κB/P65 and Survivin was significantly lower than that in the MT group(P<0. 05). The apoptosis rates of the HepG2 cells in the control group, MT group, steadily transfected group, and combination group were 3. 21%, 6.25%, 11. 82%, and 21. 06%, respectively, with significant difference found between different groups(P<0. 05). Conclusion: The apoptosis in hepatocellular carcinoma cells induced by NF-κB RNAi combined with matrine may be related to increased CD95 and Smac expression and decreased NF-κB/P65 and Survivin expression.