Abstract:
Objective To investigate the impact of miR-494 on the proliferation and invasion of human osteosarcoma cells MG-63 and to confirm whether Sox9 is the target gene of miR-494.
Methods MG-63 cells were transfected with recombinant adenovirus miR-494(Ad-miR-494), CCK-8 and clone formation experiments were employed to determine the proliferation ability of transfected MG-63 cells. Cells invasion abilities were determined by Transwell assay. Real-time PCR was used to analyze the expression of Sox9 mRNA and protein level to confirmthe adenovirus miR-494 expression in MG-63 cells. The protein expression of Sox9 was analyzed by Western blot.
Results The expression of miR-494 was increased obviously (t=36.78, P=0.000) in MG-63 cells which were transfected with Ad-miR-494. CCK-8, clone formation and Transwell experiment results revealed that the proliferation (F=1.711, P=0.012), clone formation (F=2.742, P=0.019) and invasion (F=1.653, P=0.006) abilities of MG-63 cells were markedly inhibited by the overexpression of miR-494. In addition, we observed the decreased Sox9 protein (F=5.827, P=0.021) and mRNA expression (F=5.827, P=0.021) in MG-63 cells which were transfected with Ad-miR-494. And the proliferation (t=27.54, P=0.042), lone formation (t=29.64, P=0.026) and invasion (t=32.48, P=0.016) abilities of MG-63 cells were markedly inhibited by the decreases of Sox9 expression.
Conclusion MG-63 cells proliferation and invasion abilities are suppressed by miR-494, and this process is potentially achieved via suppressing Sox9 expression.