SAHA与顺铂联合用药对骨肉瘤细胞增殖、凋亡和迁移的影响  

Effects of Suberoylanilide Hydroxamic Acid Combined with Cisplatin on the Growth, Apoptosis and Migration of Osteosarcoma Cells

作　　者：侯梦一 姚娟[1] 王豪[1] 张章[1] 严树涓[1] 左国伟[1] 

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2016年第7期777-785,共9页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81102035)资助的课题~~

摘　　要：该文研究了组蛋白去乙酰化酶抑制剂SAHA(suberoylanilide hydroxamic acid)联合顺铂(cisplatin,DDP)对骨肉瘤细胞的作用。骨肉瘤143B细胞分别经不同浓度的SAHA、DDP、SAHA+DDP作用48 h。利用倒置显微镜观察细胞形态学变化;分别采用MTT法、流式细胞术、细胞划痕实验、克隆形成实验检测SAHA、DDP及二者联用对143B细胞活力、细胞凋亡、细胞迁移和集落形成能力的影响。利用Western blot检测143B细胞中cleaved-Caspase-8、cleaved-PARP蛋白表达水平。结果显示,SAHA+DDP组143B细胞大量坏死,抑制细胞增殖的现象明显高于单药组;中低剂量SAHA与DDP联用表现为协同作用,而较高剂量的联用表现为单纯相加作用;SAHA+DDP组较单药组细胞早期凋亡率增加、迁移率明显降低,且SAHA+DDP组的细胞集落形成能力明显降低;SAHA+DDP组能够促进cleaved-Caspase-8、cleaved-PARP的蛋白表达。以上结果表明,SAHA联合顺铂对骨肉瘤细胞143B细胞有协同增敏作用,能够明显抑制其增殖和迁移并促进其凋亡。This work was aimed to investigate the effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid （SAHA） combined with DDP on osteosarcoma cells. The osteosarcoma 143B cells were treated with different concentrations of SAHA or cisplatin, either alone or combined for 48 h. The morphological characteristics of treated cells were observed by inverted microscope. The cytotoxicity effects in single drug group and combination drugs group were observed by MTT assay. The cell apoptosis was quantified by flow cytometry. SAHA or DDP was used either alone or in combination to compare the migration ability with wound healing assay. The colony-forming ability was analyzed by colony formation assay. The expressions of cleaved-Caspase-8, cleaved-PARP were detected by Western blot. The results showed that the necrosis of 143B cells was obvious in SAHA＋DDP group. The inhibition of cell proliferation in SAHA＋DDP group was significantly higher than those in single drug group. SAHA combined with DDP of middle or low doses showed a synergistic effect, however, SAHAcombined with DDP of high dose showed a simple additive effect. The early apoptosis rate of SAHA＋DDP group was significantly higher than those of single drug group, and the cell migration rate and the colony-forming ability in SAHA＋DDP group were significantly lower than single drug group. The expressions of cleaved-Caspase-8, cleaved-PARP were up-regulaton significantly in 143B cells after SAHA＋DDP treatment. The results suggested that SAHA＋DDP could inhibit the proliferation and migration, and promote the apoptosis of osteosarcoma 143B cells. In addition, the combination of SAHA and DDP has a synergistic effect.

关 键 词：SAHA 顺铂 骨肉瘤 联合用药 协同作用 

分 类 号：R738.1[医药卫生—肿瘤]