Effects of Lentivirus-mediated Cullin3 Silencing on Human Glioma Cells
QIN Yan-chang;JIA Dong;YU Qing-wei;Department of Neurosurgery,Tangdu Hospital,The Fourth Military Medical University;
Objective: To investigate the effects of Cullin3 gene silencing on the biological characteristics of human glioma cells. Methods: Cullin3 m RNA and protein levels in human glioma cells SW1783, U251 and U87 MG were detected by RT- q PCR and Western blot. A lentiviral vector expressing Cullin3 sh RNA was constructed and transfected into the U251 and U87 MG cells and the transfection efficiency was confirmed. Cells were divided into three groups: the blank(negative control group), NC-sh RNA(blank vector group) and the Cullin3-sh RNA(Cullin3-sh RNA vector group). The cell viability and cell proliferation were determined by MTT assay and Brd U incorporation assay respectively. Meanwhile, the cell migration was analyzed by Transwell assay.Flow cytometry was used to monitor the changes of cell cycle distribution. Results: The Cullin3 protein levels in SW1783,U251 and U87 MG glioma cells were significantly higher than that in the normal astrocytes. Transfection of Cullin3- sh RNA significantly abolished Cullin3 expression. Compared with the Blank and NC- sh RNA groups, Cullin3- sh RNA significantly inhibited the growth and proliferation of glioma cells U251 and U87MG.,In addtion, the migration rate was significantly decreased in Cullin3- sh RNA group. Flow cytometry indicated that the cell cycle was arrested at G0/G1 phase in Cullin3- sh RNA group. Conclusion: Cullin3 plays a critical role in gliomagenesis and development of glioma and it maybe a potential target for human glioma therapy.