Related research of the role of Autophagy gene Beclin 1 in osteosarcoma development / 中国综合临床

ABSTRACT

Objective To investigate the role of Beclin 1 in the genesis and development of osteosarcoma and the effect of Beclin 1 overexpression on the growth of the in vitro osteosarcoma cell line MG63.Methods Real time-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expressions of Beclin 1 in MG63 and hFOB1.19 at mRNA and protein levels ; A eukaryotic clone of plasmid pEGFP/Beclinl fusion with protein EGFP/Beclin 1 was constructed and was transfected into human osteosarcoma cell line MG63 by using lipofectamine 2000.The effect of Beclin1 overexpressions on the proliferation of MG63 cells was evaluated by MTT assay.Cell apoptosis was measured by flow cytomerty(FCM).Results The mRNA and protein expression of Beclin1 in human osteosarcoma cell line MG63 was significantly lower than that in the human osteoblast cell line hFOB1.19(0.17 ±0.06 vs 0.43 ±0.11,t =29.493,P ＜0.01 ; 0.13 ±0.05 vs.0.25 ± 0.08,t =6.325,P ＜ 0.01).The transfection of pEGFP/Beclinl increased the mRNA levels of human osteosarcoma(5.34 ± 0.50) times in transfected tumor cells MG63.The rate of cell apoptosis was low in control or transfected with lipofectamine 2000 only cells at an average of(0.10 ± 0.05) ％.The apoptosis rate was significantly higher in pEGFP/Beclin1 transfected cells than control cells ((4.3 ± 0.8) ％,t =5.752,P ＜ 0.05).Conclusion Compared with control cells,Bedin1 is down-regulated in the human osteosarcoma cell line MG63,which indicate the role of Beclin 1 in regulating the malignant behaviors of osteosarcoma.Beclin1 overexpressions inhibits cell proliferation and induces apoptosis in MG63 cells.