The Research Of PRIMA-1 Induce Osteosarcoma Cell Line MG-63 To Apoptosis

[Objectives]:To investigate the effect of PRIMA-1 on the proliferation and apoptosis of human osteosarcoma cell line MG-63,and further study the possible mechanism.[Method]:Coculture the P53 mutant type osteosarcoma cell MG-63 with 50μg/ml, 100μg/ml, 150μg/ml,200μg/ml,250μg/ml PRIMA-1 for 48 hours. The effect of PRIMA-1 on cell proliferation was measured by means of assy MTT , detected apoptosis induced by PRIMA-1 using Annexin V-FITC/PI flow cytometry. The change of BaxrnRNA, PUMAmRNA, bcl-2mRNA were determined by reverse transcriptase-polymerase chain reaction analysis (RT-PCR). Western bloting was performed to show the expression of P53, P53Ser15, Bax, PUMA, Caspase-3, Caspase-9, bcl-2.[RESULTS]:After coculture the P53 mutant type osteosarcoma cell MG-63 with 50μg/ml, 100μg/ml, 150μg/ml, 200μg/ml, 250μg/ml PRIMA-1 for 48 hours.the inhibitory proliferation ration of cell proliferation inhibition ratio was (3.6 ± 0.8) %, (15.2±1.2) %, (50.1±2.3) %, (63.3 ± 3.5)％, (68.7±4.3) % respectively (P<0.05) , meanwhile, the early apoptosisration of MG-63 was of MG-63 was (45.2.1±2.8) %, (36.5±3.8) %, (27.1±3.1) % (11.7±0.6) % and (3.4%±0.3) % whereas the late apoptosis ration was (11.4+1.1) % (12.6+1.9) %, (13.9+1.6) %,(24.2±2.5) % and (40.4 ± 3.6) %, respectively (P<0.05) .Theration of early apoptosis significantly decreased with increasing concentration of PRIMA-1 while ration of the late apoptosis significantly increased. Furthermore, the mRNA level of expression of Bax, PUMA were increased while the mRNA level of bcl-2 was decreased obviously. The protein levels of P53, P53Ser15, Bax, PUMA, Caspase-3, Caspase-9 was up-regulated whereas the protein levels of bcl-2 was down-regulated obviously.[CONCLUSION]:PRIMA-1 can inhibits the proliferation of MG-63 and induces apoptosis in MG-63. This may result from up-regulated expression of P53 and restoration of the conformation and function to mutant p53 protien, and relate with up-regulation of Bax, PUMA and down-regulation of bcl-2 both in mRNA level and protein level.