The Effect And Mechanism Of Silencing FBXO7 On Hepatocellular Carcinoma Cells

Objective:To investigate the effect and mechanism of silencing FBXO7 gene on the biological behavior of HepG2 and SMMC7721 cells.Methods :(1)Real-time PCR and Western blot were used to detect the expression of FBXO7 in hepatocyte L02 and 4 hepatoma cells.(2)The FBXO7 gene in HepG2 and SMMC7721 was silenced by transfecting SiRNAs liposome or lentivirus,and the negative control group was transfected with non-specific interference sequences;(3)The cell proliferation ability was detected by CCK8 method,the cell migration ability was detected by scratch test,the cell invasion ability was detected by Transwell test,the cell cycle was detected by flow cytometry,and the cell clone formation was detected by cell clone formation test;(4)Western blotting was used to detect the expression of autophagy-related proteins P62 and LC3 B,and GFP-LC3 autophagy was used to locate LC3 protein in cells.Results:(1)The results of Real-time PCR and Western blot showed that the mRNA and protein levels of hepatoma cell lines FBXO7 were higher than those of L02;(2)Real-time PCR and Western blot analysis showed that the expression of FBXO7 on mRNA and protein of HepG2/FBXO7/SiRNA cell line and SMMC7721/FBXO7/SiRNAs cell line were lower than that of the negative control groups(P < 0.05);(3)MTT cell proliferation assay and plate colony formation assay showed,compared with NC group,that the silencing groups had lower proliferation ability(P < 0.05)and fewer colony formations in vitro(P < 0.05).(4)Migration experiments and invasion experiments showed,compared with NC group,that the migration distance of the silencing groups were shorter(P < 0.05)and the number of transmembrane cells was fewer(P < 0.05).(5)Flow cytometry showed,compared with NC group,that the silencing groups had higher proportion of cells in the G0 phase and lower proportion of cells in the S phase(P < 0.05).(6)Western blotting results showed,compared with NC group,that the P62 protein of the silencing groups decreased and the LC3 B protein increased.(7)Compared with NC group,the percent of LC3 puncta positive per cells was increased in silencing groups by transfecting GFP-LC3 adenovirus(P < 0.05).Conclusion:(1)The expression of FBXO7 in hepatoma cell lines was higher than that in L02;(2)Silencing FBXO7 inhibits proliferation,invasion and migration of hepatoma cell lines;(3)Silencing FBXO7 blocks hepatoma cell lines in G0 phase,reducing the proportion of cells in S phase;(4)Silencing FBXO7 promotes autophagy hepatoma cell lines.