三氧化二砷诱导骨肉瘤细胞凋亡的实验研究  

Arsenic trioxide induced osteosarcoma cell apoptosis in vitro

作　　者：方成[1] 陈振光[1] 喻爱喜[1] 祝少博[1] 

机构地区：[1]武汉大学中南医院显微骨科,430071

出　　处：《中华骨科杂志》2003年第6期345-348,共4页Chinese Journal of Orthopaedics

摘　　要：目的研究三氧化二砷(As2O3)诱导骨肉瘤细胞凋亡的生物学效应及其机制。方法应用MTT(methythiazoloyltetrazolium)法、倒置相差显微镜观察、荧光染色、原位末端标记(TUNEL)法、流式细胞仪和RT-PCR(reverse-transcriptasepolymerasechainreaction)检测,观察As2O3对骨肉瘤细胞株MG-63诱导凋亡的作用。结果As2O3可明显抑制MG-63骨肉瘤细胞的增殖,1μmol/L以上浓度的As2O3对MG-63骨肉瘤细胞的抑制率可达70%。As2O3抑制MG-63骨肉瘤细胞增殖主要通过诱导细胞凋亡实现。倒置相差显微镜及荧光染色观察被As2O3抑制的MG-63细胞呈典型的凋亡改变;大部分细胞TUNEL法检测阳性;流式细胞仪检测凋亡率与As2O3处理时间、处理浓度呈正相关,细胞周期明显阻滞于G2/M期;RT-PCR显示As2O3处理后原癌基因c-myc表达降低。结论As2O3可有效地诱导骨肉瘤细胞凋亡,c-myc表达降低是诱导MG-63骨肉瘤细胞凋亡的重要机制。Objective The clinical results of leukemia improved significantly due to the application of arsenic trioxide(As 2 O 3 ).This compound was used in inducing apoptosis on osteosarcoma cell lines and its mech anism was explored.Methods Human osteosarcoma MG-63cell line was cultured and treated with0.25,0.5,1,2,4and8μmol/L As 2 O 3 .The cell line was measured at24,48,72,96and120h after As 2 O 3 infiltration.MTT assay,phase contrast light microscopy,fluorescence staining,TUNEL,flow cytometry anal-ysis and RT -PCR were used to investigate the inducing apoptosis and inhibitative effect of As 2 O 3 on os-teosarcoma MG-63cell line.Results As 2 O 3 could inhibit the reproduction of osteosarcoma cell line MG-63significantly,the inhibitive rates of MG-63cell line treated with As 2 O 3 over1μmol/L reached70%.Un -der phase contrast light microscopy and fluorescence staining,osteosarcoma cells cultured with As 2 O 3 pre sent -ed typical apoptotic changes.The majority of cells cultured with As 2 O 3 were positive with TUNEL.Flow cy tometry analysis showed a time-dose dependent apoptosis rate and a remarkable G 2 /M phase arrest.The ex pression of c-myc was decreased in MG-63cells treated with As 2 O 3 .Conclusion As 2 O 3 is able to in duce osteosarcoma apoptosis effectively.The expression of decreasing c-myc is an important mechanism in induc-ing osteosarcoma MG-63cells apoptosis.

关 键 词：骨肉瘤细胞 AS2O3 凋亡 诱导 三氧化二砷 C-MYC表达 显微镜观察 浓度 处理时间 荧光染色 

分 类 号：R738.1[医药卫生—肿瘤]