Expression And Significance Of Annexin A3 In Osteosarcoma HOS Cells

Objective: To investigate the difference and significance of Annexin A3(ANXA3)expression in osteosarcoma HOS cells and osteoblast cells,and to seek a new targeted therapy for osteosarcoma.Methods: The expression of ANXA3 in osteosarcoma cells was confirmed by immunofluorescence test at first.Reverse transcription-polymerase chain reaction(RT-PCR)and Western blot were used to detect ANXA3 in osteoblast cells and osteosarcoma HOS and U2 OS cells respectively.Transfecting small interfering RNA(siRNA)into HOS cells by liposome-mediated method so that the expression of ANXA3 in HOS cells was down-regulated.We observe fluorescence by fluorescence microscope after 6 hours.Total protein was extracted after transfecting 48 hours and we find ANXA3 in the osteosarcoma HOS cells was down-regulation by Western blot.Then,the apoptosis rate of HOS cells before and after transfection was detected by flow cytometry.Results:1.The expression of ANXA3 in osteosarcoma cells was observed by confocal laser scanning microscope;2.The expression of ANXA3 in osteoblast cells and osteosarcoma cells were detected by RT-PCR and Western blot.It is found that ANXA3 in osteosarcoma HOS and U2 OS cells were significantly higher than in osteoblast cells both in RNA and protein expression levels,especially more significantly in osteosarcoma HOS cells;3.After transfecting the three siRNAs into osteosarcoma HOS cells,the transfection efficiency of osteosarcoma HOS cells were detected by Western blot.The result showed that the expression of ANXA3 was down-regulated in each experimental group;4.The apoptosis of osteosarcoma HOS cells before and after transfection were detected by flow cytometry.The result showed that Apoptosis rate increased significantly after transfection compared with before transfection.Conclusion: ANXA3 is highly expressed in osteosarcoma cells.Down-regulation of ANXA3 in osteosarcoma HOS cells can increase apoptosis.