组蛋白去乙酰化酶抑制剂对人骨肉瘤细胞生长的影响  

Histone deacetylase inhibits the growth and migration of human osteosarcoma cells

作　　者：张昭亮 张爱华[1] 封志强[1] 安利[1] 赵庆华[2] 

机构地区：[1]山东省泰安市妇幼保健院外科,271000 [2]上海交通大学附属第一人民医院骨科

出　　处：《中华医学杂志》2013年第33期2680-2683,共4页National Medical Journal of China

摘　　要：目的观察组蛋白去乙酰化酶抑制剂（HDACIs）在体内及体外对人骨肉瘤细胞的作用机制。方法本实验应用两种不同类型的HDACIs：辛二酰基酰替苯胺异羟肟酸（SAHA）和丁酸钠（SB），评价其在体内和体外对人骨肉瘤细胞（SaOS2、U2OS）生长的影响。应用甲基甲苯基硫细胞检测（MTS）法检测细胞增殖活性，流式细胞技术和蛋白免疫印迹法检测细胞周期和组蛋白乙酰化程度。结果MTS显示SAHA及sB浓度由0，0．04μmol／L每5倍递增，最大剂量625μmol／L，SaOS2A值分别由1．8降至0．46，1．8降至0．7；U20S细胞A值分别由1．62降至0．37，1．63降至0．53。SAHA和SB对SaOS，和U2OS细胞生长的抑制具有明显的浓度依赖性。蛋白免疫印迹法显示HDACIs治疗后，组蛋白H3乙酰化水平上升，SAHA处理后SaOS2和U2OS细胞的乙酰化H3与对照组组蛋白乙酰化水平比值分别为1．4和1．7；SB处理后的比值分别为1．2和1．3。流式细胞技术显示SAHA将细胞阻滞在分裂周期的G1和G2／M期，而sB将细胞阻滞在分裂周期的G2／M期。同时，肿瘤抑制因子的效果在小鼠动物模型上得到了有效的验证，SAHA组（2．0mg／kg）第1天肿瘤体积103．8mm^3，28d后相比对照组明显减小（1040mm^3比1465mm^3）；SB组（50mg／kg）第1天肿瘤体积103．0mm^3，第28天后相比对照组明显减小（1040mm^3比1465mm^3）。结论SAHA和SB能够阻滞小鼠模型体内的SaOS2细胞。SAHA和SB可抑制了人骨肉瘤细胞生长，阻碍了细胞周期进程。Objective To explore the effects of histone deacetylase inhibitors （HDACIs） on human osteosarcoma in vitro and in vivo. Methods HDACI suberoylanilide hydroxamic acid （SAHA） and sodium butyrate （SB） were employed to examine the effects on human osteosarcoma in vitro and in vivo. The in vitro effects of HDACI SAHA and SB were evaluated in SaOS2 and U2OS human osteosarcoma cell lines. Cell growth, cell cycle progression and histone acetylation were examined by methyl tolyl sulfide （ MTS）, flow cytometry and Western blot respectively. In addition, SAHA or SB was administered for 4 weeks in a murine xenograph model for assessing the in vitro effects. Results MTS assay revealed that SAHA and SB significantly suppressed the growth of SaOS2 and U2OS cells in a concentration-dependent manner. Western blot analysis indicated that the levels of acetylated H3 increased alter HDACI treatment. Flow cytometry showed that SAHA arrested the cell cycle in G, and GJM phase, while SB arrested the cell cycle in G2/M phase. The tumor growth of murine xenograph model with SaOS2 was inhibited by SAHA and SB compared with vehicle control. Conclusion HDACI SAHA and SB significantly inhibit the growth of human osteosarcoma cells and induce cell cycle arrest. Meanwhile, the tumor inhibitory effects were also validated in a routine xenograft model.

关 键 词：骨肉瘤 组蛋白脱乙酰基酶 酶抑制剂 细胞周期 

分 类 号：R738[医药卫生—肿瘤]