Effects Of SiRNA-targeting BMP-7 On The Proliferation And Migration Of Human Liver Cancer HepG2 Cells

Purpose:To investigate the effect of small interfering RNA(siRNA)targeting bone morphogenetic protein 7(BMP-7)on biological activity(cell proliferation,cell migration and cell cycle)of human liver cancer cells.Methods:The m RNA and protein levels of BMP-7 in cells of Hep G2 and SMMC7721 were determined with reverse transcript-polymerase chain reaction(RT-PCR)and western blot and the highest expressed cell was selected for subsequent experiment.Simultaneously,three pairs of siRNA targeting BMP-7and negative siRNA were synthesized and transiently transfected into liver cancer cells by Trans Lipid? HL Transfection Reagent.We constructed five groups including a normal control group,a negative-siRNA group and 3 transfected groups with BMP-7-siRNA-1,BMP-7-siRNA-2,BMP-7-siRNA-3.The m RNA and pritein levels of BMP-7 was measured by RT-PCR and western blot,respectively.And the best sequence for BMP-7 silence is selected.The proliferation and migration of transfected cells were assessed using MTT assay and transwell migration assay,respectively.The expression of apoptosisrelated protein(BAX,Bcl-2,Caspase-3)in each group were determined by Western blotting,and the cell cycle was analyzed by flow cytometry.Results:Western blot and RT-PCR indicated that Hep G2 was the highest expression of BMP-7 in the two liver cancer lines(P<0.01).BMP-7-siRNA-3 group demonstrated the lowest level among the five groups(P<0.01).It revealed that the number of cell growth were significantly lower in BMP-7-siRNA-3 group than in control groups 48 h and 72 h after cells were transfected(P<0.01).The numbers of cell transmembrane were significantly lower in BMP-7-siRNA-3 group than in control groups 24 h after cells were transfected(P<0.01).The expression of Bax and Caspase-3(P<0.01)were significantly increased after BMP-7 silence and there was no statistically significant difference about the Bcl-2 expression by western blot.Flow cytometer showed that cells were significantly blocked in G2 phase 48 h after cells were transfected in BMP-7-siRNA-3 group(P<0.05).Conclusions:siRNA-targeting BMP-7 can markedly inhibited the expression of BMP-7 in liver cancer Hep G2 cells,and decrease the abilities of proliferation and migration,promote the abilities of cell apoptosis and block G2 phase of cell cycle of liver cancer cells,especially in BMP-7-siRNA-3 group.