As_2O_3对人骨肉瘤细胞的体外效应

Effect of arsenic trioxide on human osteosarcoma cells in vitro

目的研究三氧化二砷(As2O3)对人骨肉瘤细胞U20S的生物学效应及其机制。方法利用MTT法观察As2O3对U20S 细胞的生长抑制作用;Annexin V-FTTC+PI双参数检测细胞凋亡;流式细胞仪测定经不同浓度As2O3处理后U20S细胞增殖细胞核抗原(PCNA)、凋亡相关蛋白Fas和Bcl-2阳性细胞百分率及细胞内钙离子(IECa2+)含量变化。结果As2O3可显著抑制U20S细胞生长,且剂量-效应和时间-效应关系显著(P<0.05),其24,钙和72 h的IC50值分别为10.66,2.43和0.46 μmol·L-1。U20S细胞经As2O3处理后可发生凋亡。As2O3能显著增加Fas蛋白表达和IECa2+含量(P<0.01),下调PCNA蛋白表达(P< 0.01),对Bcl-2表达无影响(P>0.05)。结论As2O3在体外可显著抑制人骨肉瘤细胞U20S生长并引起凋亡,其机制可能与上调Fas表达和IECa2+含量及降低PCNA蛋白表达有关。

OBJECTIVE To research the biological effect of arsenic trioxide on human osteosarcoma cell line U20S and its mechanism.METHODS MTT method was used to evaluate the growth inhibition effect. Apoptosis was detected with Annexin V-FITC ＋ PI dual parameter. The positive rate of proliferation cell nuclear antigen （PCNA） , apoptosis associated protein Fas and Bcl-2, intracellular calcium ions （ IECa^2 ＋ ） levels were measured by flow cytometry in U20S ceils treated with different doses of As2O3. RESULTS As2O3 inhibites the growth of U20S ceils dramatically. A obvious dosage-effect and time-effect correlation were obtained, its half inhibitory concentrations （IC50） at 24,48 and 72 h were 10.66,2.43 and 0.46μmol·L^-1 ,respectively. Apoptosis occurred in U20S ceils was treated with As2O3. As2O3 increased Fas protein expression and IECa^2＋ levels, and decrease PCNA protein expression in U20S ceils（ P 〈 0.01 ）. Bcl-2 protein expression wasn＇t affected （ P 〉 0.05 ）. CONCLUSION As2O3 inhibites the growth of U20S cells in vitro dramatically and induces apoptosis. Its mechanism is probably associated with the improve Fas expression and IECa^2＋ levels, and decreases PCNA protein expression.