Transfection of PIN1 antisense oligonucleotides inhibited proliferation of human osteosarcoma cells

Objective:To evaluate the inhibitory effects of PIN1 antisense oligonucleotides on the poliferation of human osteosarma cells. Methods:Different doses of antisense PIN1 oligonucleotides (0,20, 50,100,200,250 μL) in plasmid pIRES2-EGFP vector were transfected into osteosarcoma MG-63 cells. The cultured cells and supernatant before and after transfection were collected. The cell proliferation was determined by MTT method. The cell cycle and apoptosis were detected by FCM. The PIN1 protein expression was detected by Western blot and its mRNA expression was detected by reversed transcriptional polymerase chain reaction (RT-PCR). Results:MTT and FCM assays indicated that the transfection with antisense PIN1 oligonucleotides could inhibit proliferation of MG-63 cells and lead to cell apoptosis. Western blot analysis revealed that antisense PIN1 oligonucleotides inhibited PIN1 protein expression in a dose-dependent manner ( P 0.05). The optical density was 0.854±0.136, 0.866±0.138 ,0.732±0.154,0.611±0.121,0.547±0.109,0.398±0.113,0.320±0.151 after transfection of 0,20,50,100,200,250 μL of antisense PIN1 oligonucleotides. Conclusion:The expression of PIN1 mRNA in MG-63 cells could be inhibited by antisense PIN1 oligonucleotides which reduced the expression of PIN1 protein and suppressed the proliferation of human osteosarcoma MG-63 cells.