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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 To study the biological effects of arsenic trioxide (As 2 O 3) on osteosarcoma cell apoptosis and its mechanism. Methods Methyl thiazoloyl tetrazolium (MTT) assay, inverted phase contrast microscope, fluorescent staining, TUNEL, flow cytometry and reverse transcriptase polymerase chain reaction (PCR) were used to detect the effect of As2O3 on osteosarcoma cell line MG-63 Induction of apoptosis. Results As2O3 could significantly inhibit the proliferation of MG-63 osteosarcoma cells. The inhibitory rate of As2O3 on MG-63 osteosarcoma cells was up to 70% when the concentration of As2O3 was above 1μmol / L. As2O3 inhibits proliferation of MG-63 osteosarcoma cells primarily by inducing apoptosis. Inverted phase contrast microscope and fluorescent staining were used to observe the typical apoptotic changes of MG-63 cells treated with As2O3. Most of the cells were detected by TUNEL method. Flow cytometry showed that the apoptosis rate was positively correlated with the treatment time and concentration of As2O3, Obviously blocked in G2 / M phase; RT-PCR showed that As2O3 treated c-myc gene expression decreased. Conclusion As2O3 can effectively induce the apoptosis of osteosarcoma cells. The decrease of c-myc expression is an important mechanism of inducing MG-63 osteosarcoma cell apoptosis.