组蛋白去乙酰化酶抑制剂PCI-24781诱导耐甲氨蝶呤骨肉瘤细胞U2-OS／MT300凋亡  

Histone Deacetylases Inihibitor PCI-24781 Induces Apoptosis in Methotrexate Resistant Cell Line U2-OS/MTX300 in Vitro

作　　者：尹军强[1] 谢显彪[1] 邹昌业[1] 王晋[1] 黄纲[1] 谭平先[1] 雍碧城[1] 沈靖南[1] 

机构地区：[1]中山大学附属第一医院肿瘤科.广州510080

出　　处：《中国药学杂志》2012年第20期1630-1633,共4页Chinese Pharmaceutical Journal

基　　金：广东省自然科学基金资助项目(8251008901000019)

摘　　要：目的研究组蛋白去乙酰化酶抑制剂PCI-24781诱导耐甲氨蝶呤骨肉瘤细胞凋亡的作用及机制。方法四甲基偶氮唑蓝(MTT)法及平板克隆形成实验检测PCI-24781对骨肉瘤细胞U-20S/MTX300增殖的影响,hochest33258染色检测PCI-24781处理后细胞核形态改变,流式细胞仪检测凋亡,Western blot法检测cleaved-PARP、P53表达量及组蛋白H3、H4乙酰化水平变化。结果PCI-24781对耐甲氨蝶呤U2-OS/MT 300细胞呈剂量依赖性增殖抑制,其48 h IC50值为(0.55±0.03)μmol.L-1,该值与甲氨蝶呤敏感U2-OS细胞IC50值之间无统计学差异(P>0.05)。0.5μmol.L-1PCI-24781可显著抑制细胞克隆形成,其抑制率达(61±7)%。PCI-24781处理48 h后细胞内出现典型凋亡小体,流式细胞仪检测凋亡率达(29±4)%。Western blot检测可见随PCI-24781浓度升高cleaved-PARP,组蛋白H3、H4乙酰化水平及P53显著升高。结论组蛋白去乙酰化酶抑制剂PCI-24781可显著抑制耐甲氨蝶呤骨肉瘤细胞增殖并诱导细胞凋亡,其作用机制与上调组蛋白乙酰化水平,促进抑癌基因P53表达有关。OBJECTIVE To study the antitumor activity and the related mechanism of histone deacetylase inihibitor PCI-24781 on methotrexate resistant osteosarcoma cell line, U2-OS/MTX300. METHODS U-20S/MTX300 ceils were treated with PCI-24781. Cell viability was evaluated by MTT and colony formation assay. Apoptosis was evaluated by flow cytometry and fluorescent staining. Western Blot was used to detect the expression of apoptosis-related proteins and the acetylated level of histone 3 and 4. RESULTS PCI-24781 inhibited the viability of U-20S/MTX300 in a concentrationdependent manner. The IC50 value of PC1-24781 on U-20S/ MTX300 at 48 h ＇after administration was （0. 55 -＋ 0. 03 ） μmol · L^-1 , which was similar to its IC50 value in the MTX sensitive cells （ P 〉 0. 05）. 0. 5 μmol · L^-1 PCI-24781 treatment could inhibit the colony formation obviously, and the colony inhibit rate was （61 - 7 ）%. Typical apoptotic bodies were observed at 48 h after PCI-24781 treatment and the apoptosis rate was （29 ± 4）%. PCI-24781 could induce up-regulation of cleaved-PARP and P53. The acetylated level of histone 3 and 4 were also significantly increased after PCI-24781 treatment. CONCLUSION Histone deacetylases inihibitor PCI-24781 could inhibit the growth and induce apoptosis in methotrexate resistant osteosarcoma cells. Up-regulation of acetylated histone proteins and P53 are involved in this process.

关 键 词：组蛋白去乙酰化酶 抑制剂 骨肉瘤 细胞凋亡 甲氨蝶呤 

分 类 号：R965[医药卫生—药理学]