Expression and significance of annexin A3 in osteosarcoma HOS cells

Abstract

Objective To investigate the expression and significance of annexin A3 (ANXA3) in osteosarcoma HOS cells. Methods The expression levels of ANXA3 in osteoblasts cells, osteosarcoma HOS cells and osteosarcoma U2OS cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting respectively. The expression of ANXA3 in HOS cells was down-regulated by transfection of Small interfering RNA (siRNA) into HOS cells by liposome-mediated method. Flow cytometry was used to detect the apoptosis of cells before and after transfection. Results Compared with the osteoblast control group, the expression of RNA and protein in ANXA3 cells was significantly increased in HOS and U2OS cells. At the level of RNA, HOS cells was 2.99 times higher than that of osteoblasts cells, and U2OS cells was 1.25 times higher than that of osteoblasts cells. At the protein level, HOS was 3.25 times higher than that of osteoblasts, and U2OS was 2.33 times higher than that of osteoblasts. siRNA down-regulated ANXA3 expression 48 hours and Western blotting detection transfection efficiency, Compared with the control group, the down-regulation of the si-651 group was 0.26, the down-regulation of the si-732 group was 0.31, the down-regulation of the si-1094 group was 0.31, the positive control group was 0.37, the negative control group was 0.87. The apoptotic rate was 8.1% in the control group and 21.5% in the si-651 group, which was 15.2% higher than that in the control group. The apoptosis rate of si-732 group was 13.3%, which was 7.5% higher than that of control group. Compared with the control group increased 7.5%; si-1094 group apoptosis rate was 13.4%, compared with the control group increased 6.7%. Conclusion The expression of ANXA3 in osteosarcoma cells is improved; Down-regulation of ANXA3 expression in HOS cells can increase the apoptotic capacity. Key words: Osteosarcoma; Annexin A3; Gene silencing; Targeted therapy