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.
ABSTRACT
Objective To investigate the effective regimen to treat the hepatitis C virus (HCV) recurrence after orthotopic liver transplantation (OLT). Methods The clinical data of 4 cases of HCV recurrence after OLT were retrospectively analyzed. Of the 4 cases, there were 3 cases of HCV related liver cirrhosis and 1 case of HCV related liver cirrhosis in combination with hepatocellular carcinoma. The immunosuppression regimen as FK506, MMF and corticosteroids was used after OLT. As soon as HCV recurrence was confirmed by liver biopsy during 8 to 12 weeks after OLT, pegylated interferonα-2a (PEG-IFNα-2a) and ribavirin (RIB) were used for 48 weeks. PEG-IFNα-2a was started at a dose of 180 μg per week subcutaneously and RIB at a dose of 1000 mg per day orally, respective-ly. Blood routine, liver and kidney function test, HCV-RNA and transplanted liver biopsy were per-formed when necessary and biochemical response, sustained virologic response and histological re-sponse were tested in due time. Remits All of the 4 cases except for 1 achieved sustained virologic re-sponse and the liver function was as normal as before. The histological activity index was improved significantly for both inflammatory activity and fibrosis according to liver biopsy in 0, 48, 72 week srespectively. Case 4 was given corticosteroids for consecutively 3 days when acute rejection was veri-fied by liver biopsy and the condition improved. None of them stopped treatment or withdrew from them directly. Conclusion The combination of PEG-IFNα-2a and RIB was an effective regimen to treat the HCV recurrence after OLT and the side effects could be overcame easily.