| Background and objective:China has a large number of hepatitis.Hepatocellular carcinoma(HCC)is a common tumor with complicated pathogenesis.Heredity,diet,virus and parasite infection may be related to the occurrence and development of liver cancer.With the development of molecular biology,a variety of liver cancer related genes have been studied by different researchers,and was thought to be involved in the pathogenesis of HCC.Araujo et al.indicated that QSOX1 was highly expressed in tumor tissues of highly differentiated neuroblastoma.They found that QSOX1 was also highly expressed in patients with a high prevalence of relapses/residual disease.Furthermore,they suggested that QSOX1 played an important role in the differentiation and invasion of neuroblastoma.QSOX1 was also thought to be involved in the development of breast and prostate cancer.At present,the research on therelationship between QSOX1 and the occurrence and development of liver cancer is limited.In this study,we further study the expression of QSOX1 in hepatocellular carcinoma cells and its effect on the biological behavior of hepatocellular carcinoma cells,and to reveal the role of QSOX1 in the biological function of hepatocellular carcinoma cells,which may provide theoretical and experimental basis for early diagnosis and gene therapy of liver cancer.Methods : The lentivirus vector interfering with QSOX1 was constructed.The lentivirus vector and empty vector were transfected into SMMC-7721 hepatoma cells,respectively.The vacuity contrast group was done without any disposal.The stable expression strains were screened out,and the culture was extended.The real-time quantitative PCR and western blot were used to detect the expression level of mRNA and protein of QSOX1 gene in human SMMC-7721 hepatoma cells,respectively.The CCK8 assay was used to verify the proliferation of SMMC-7721 cells.The colony formation assay was applied to determine the tumorigenesis ability.Flow cytometry was used to examine the cell cycles of SMMC-7721 cells.Transwell assay and invasion experiment were implemented to detect the ability of metastasis and invasion of SMMC-7721 cells.Results : According to the multiplicity of infection,both the experimental and the control group were successfully transfected.Theexpression of QSOX1 gene was significantly inhibited in the transfected group when compared with the empty vector and vacuity contrast groups(P < 0.05).After culture and treatment with CCK-8,the proliferation of lentivirus transfection group was significantly slower than that of other control groups(P < 0.05).The number of cell clones in lentivirus transfection group was significantly lower than that in other control groups(P < 0.05).The flow cytometry showed that the percentage of G1 cells in the lentivirus transfection group was significantly lower than that in the empty cell group(P < 0.001),While the S phase increased significantly,and no significant changes in the G2/M period.When compared with the empty virus transfection group,the G1 cells of the lentivirus transfection group were significantly reduced,the S phase cells did not change significantly,while the G2/M phase cells increased significantly.The cell proliferation and tumor forming ability of the transfected group were significantly lower than that of the other control groups(P < 0.05).The migration rate and metastasis rate of the transfected cells were lower than those of untransfected cells(P < 0.05).Conclusions:By lentivirus interference QSOX1 gene expression in SMMC-7721 cells in vitro,the proliferation and tumorigenicity of HCC cells were significantly decreased.Transfer and migration ability decreased significantly.The gene QSOX1 may play an important role inpromoting cell growth,proliferation,invasion and metastasis of SMMC-7721 hepatoma cells.The results provide a basis for the research of gene therapy for hepatoma carcinoma. |
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