| Objective:According to the result of a gene-array analysis in our preliminary experiment, it comes to a conclusion that GTPBP4 expresses highly in hepatocellular carcinoma.Our study is aimed at making it clear that whether and how GTPBP4 influences on the migration and invasion, by silencing GTPBP4 with lentiviral vector-mediated RNA interference.Methods:1.At first, we picked out a most effective GTPBP4-silenced lentiviral vector using Quantitative Real Time-PCR for mRNA level and western blotting for protein level in human HCC cell lines SMMC-7721 and MHCC97-H, followed by verifying the silence of GTPBP4 expression.2.With the selected GTPBP4-silenced lentiviral vector infecting human HCC cell lines SMMC-7721 and MHCC97-H, we detected the migration by transwell invasion assay and the invasion by wound scratch assay. There were three groups including Normal group, Negative Control group, Experiment group.3.The protein levels of MMP-9 were examined by western blotting after GTPBP4-silence.Results:1.Western blot and qRT-PCR uncovered combinatively that the expression of GTPBP4 at the level of mRNA and protein in NO.1, NO.2 and NO.3 group were silenced, obviously in NO.1 group.2.The migration was blocked in Experiment group in SMMC-7721 and MHCC97-H, while Normal group and Negative Control group remains no statistical difference.3.The invasion was blocked in Experiment group in SMMC-7721 and MHCC97-H, while Normal group and Negative Control group remains no statistical difference.4.The protein levels of MMP-9 were downregulated with GTPBP4-silence inSMMC-7721 and MHCC97-H.Conclusions:The migration and invasion were inhibited obviously in SMMC-7721 and MHCC97-H after the selected GTPBP4-silenced lentiviral vector infecting human HCC cell lines. Meanwhile, the protein levels of MMP-9 were downregulated.Therefore, we speculate that GTPBP4 probably participate in the migration and invasion in human hepatocellular carcinoma. |
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