Effect of GPR49 gene on proliferation and invasive ability of hepatoma cells / 器官移植

ABSTRACT

Objective To discuss the effect of G-protein-coupled receptor 49 (GPR49)gene on proliferation and invasive ability of hepatoma cell line Huh7 and its molecular biological mechanism.Methods According to the different transfected small interfering RNA(si-RNA),Huh7 cells were divided into the GPR49-siRNA(si-GPR49)group and the NC-siRNA (si-NC)group.Untransfected Huh7 cells were set as the control group. Messenger RNA (mRNA )and protein expression of GPR49, cyclin D1 and matrix metalloproteinase 9 (MMP9)in the cells of the three groups were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR)and Western blot method.The proliferation and invasive ability of the cells of each group were respectively detected by MTT method and Transwell method.Results The relative expression of GPR49 mRNA of the si-GPR49 group was (23.8 ±3.1)% of the control group (P <0.05). Compared with the control group,the protein expression of GPR49,cyclin D1 and MMP9 of the si-GPR49 group decreased significantly (all in P <0.05).The proliferation experiment results by MTT indicated that the optical density(OD)of the cells of the si-GPR49 group at 72 h was (0.53 ±0.12),which was significantly lower than that of the control group (1.35 ±0.28).The difference had statistical significance (P <0.05). The average invaded cell counts of the si-GPR49 group were (13.6 ±2.5),which was significantly lower than (65.3 ±6.1 )of the control group.The difference had statistical significance (P <0.05 ).Conclusions GPR49-siRNA may inhibit the gene expression of GPR49 in Huh7 cells.Its mechanism may be that the proliferation of Huh7 cells is inhibited by reducing the level of cyclin D1;the migration and invasive ability of Huh7 cells is inhibited by affecting the expression level of MMP9.