| Objective:To investigate the influence of P27RF-Rho m RNA gene silencing on the drug sensitivity of 5-Fu to SMMC7721 cell line,and to provide theoretical basis for the treatment of advanced liver cancer.Methods:P27RF-Rho RNAi vector and P27RF-Rho gene silencing lentivirus were used to infect the SMMC7721 cells.Western blotting method was used to detect the gene silencing effect.The SMMC7721 cells were divided into Scramble-si RNA group,5-Fu group,P27RF-Rho-si RNA group and P27RF-Rho-si RNA + 5-Fu group.Western blotting was used to detect the infection efficiency of RNAi.MTT method was used to detect the cell growth in various groups.Scratching test was used to detect the migration ability of cells in various groups.Transwell experiment were used to detect the invasion ability of cells.The expressions of P27 and Rho C protein were detected by Western blotting method.Results:P27RF-Rho RNAi lentiviral vector was successfully constructed.Fluorescence microscopy showed that lentivirus infection was good.TheWestern blotting results showed that the expression of P27RF-Rho protein in P27RF-Rho-si RNA group was decreased compared with SMMC7721 group and Scramble-si RNA group.Compared with three control groups,the cell growth speed of the cells in P27RF-Rho-si RNA +5-Fu group was significantly decreased(P<0.05).The cell migration ability of the cells in P27RF-Rho-si RNA + 5-Fu group was significantly lower than those in other three groups(P <0.01);the average number of cells passing through the Transwell microporous membrane was significantly less than other groups(P<0.01);The Western blotting analysis results showed that the expression level of P27 protein in the cells in P27RF-Rho-si RNA + 5-Fu group was significantly higher than those in other three groups(P<0.05);The expression of Rho C protein was significantly lower than those in other three groups(P<0.05).Conclusion: P27RF-Rho gene silencing can significantly enhance the drug sensitivity of 5-Fu to SMMC7721 cell. |
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