| Objective:Explore the effect of inhibiting the expression of PDGFR-β by RNA interference on proliferation and radiosensitivity in rat C6glioma cells.Methods:Build the PDGFR-p-shRNA-Lentiviral RNAi vector and then transfected into rat C6glioma cells. The cells were divided into the NC(Negative Control,NC)group and KD (Knock Down,KD) group.Western-blot detection test the suppression level of PDGFR-β protein expression in C6cells. The growth curve of the two groups were compared with MTT assay. Clonogenic assay test the cell survival rate and the radiosensitivity after two groups combined with radiotherapy. Flow cytometry analyse the cell cycle distribution of two groups combined with radiotherapyResults:Lentiviral vector transfection of C6cells were observed under fluorescence microscope, fluorescence ratio is greater than80%, showed that the successful transfection. Western-blot showed the expression of PDGFR-β in KD group level decreased significantly, and compare with the NC group had significant differences(P<0.05), protein expression inhibition rate was83.4%. Cell proliferation in KD group was slightly inhibited, but compared with the NC group had no statistical significance by MTT assay. Clonogenic assay displayed the mean lethal dose(Do) in the KD group combined with radiotherapy decreased from1.843Gy to1.297Gy compared with the NC group, and the radiosensitization ratio(SER)=1.421. Flow cytometry showed cell cycle arrest in the G2/M phase in KD group combined with radiotherapy.Conclution:Silence of PDGFR-β expression resulted from RNA interference, can not inhibit the rat C6gliomas cells proliferation, and enhance the radiosensitivity of C6gliomas cells in vitro. PDGFR-β can be used to treat glioblastoma as one of the targets. |
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