Objective: To explore the effect of miR-27a knockdown on U87 glioma cells in vitro. Methods: The lentiviral vector miRZip-27a anti-miR-27a microRNA construct,empty lentiviral vector pGreenPuro Scramble Hairpin Control-Construct,and lentiviral packaging plasmids in supernatant were collected and transfected into 293T cells,the packaged lentivirus were named as Lt-I and Lt-NC. Then the packaged lentivirus Lt-I and Lt-NC were used to infect U87 glioma cells,the stable infected U87 glioma cells were selected by flow cytometry(FCM) for the next research. Then quantity-PCR,CCK-8,and transwell invasion assay were used to detect the effects of miR-27a knockdown. Results: Compared with the uninfected U87 glioma cells,the stable infected U87 glioma cells by Lt-I had significantly decreased miR-27a expression,cell proliferation and invasiveness. These changes were not observed in the U87 glioma cells stably infected with Lt-NC. Conclusion: Knockdown of miR-27a can inhibit the proliferation and invasiveness of the U87 glioma cells.