Inhibitory Effect Of SiRNA On Abnormal Expression Of FAT10 In Hepatomacellular Carcinoma

Objective: Design and select siRNA sequences that could inhibit the expression of ubiquitin-related protein FAT10. Then transfect the siRNA into Hep3B cells by LipofectamineTM 2000 and research the effect and significance on biological behavior of hepatoma carcinoma cells.Methods: 1. Four siRNA sequences were designed and transfected into Hep3B cells. The tendency of FAT10 expression was estimated using RT-PCR, Real-Time PCR.Select the best siRNA and the best interfering condition. 2. Again transfect the best siRNA into Hep3B cells via the best interfering condition. Immunocytochemical method was used to detect the change of FAT10. 3. To detect the influence on biological behavior of Hep3B after transfection. The effect on cell growth was detected by MTT and the effect on the change of cell cycle was analyzed by flow cytometry when FAT10 was degraded by siRNA.Results: 1. Transfection efficiency was demonstrated high by observing the green fluorescence through fluorescence microscope. 2. The expression of FAT10 was degraded obviously by the detection of RT-PCR, Real-Time PCR and immunocytochemical method.3. The cell growth ability of Hep3B cells was markedly inhibited following FAT10 mRNA degraded by siRNA transfection. Hep3B cells were mainly detained in G0/G1 and the percentage of S and G2/M was decreased.Conclusion: siRNA can effectively inhibit abnormal expression of FAT10 as well as growth of Hep3B.FAT10 may play an important role in the occurring mechanisms of hepatocellular carcinoma.