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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。
In this paper, the effect of histone deacetylase inhibitor SAHA (suberoylanilide hydroxamic acid) and cisplatin (DDP) on osteosarcoma cells was studied. Osteosarcoma 143B cells were treated with different concentrations of SAHA, DDP and SAHA + DDP for 48 h. Cell morphological changes were observed by inverted microscope. Cell viability, apoptosis, cell migration and colony formation of 143B cells were detected by MTT, flow cytometry, cell scratch assay and clone formation assay, respectively The impact of ability. The protein level of cleaved-Caspase-8 and cleaved-PARP in 143B cells was detected by Western blot. The results showed that the SAH + DDP group of 143B cells necrosis, inhibition of cell proliferation was significantly higher than the single drug group; low-dose SAHA and DDP showed synergy, while the combination of higher doses showed a simple additive effect The apoptosis rate of SAHA + DDP group was significantly higher than that of SAHA + DDP group, and the migration rate of SAHA + DDP group was significantly lower than that of SAHA + DDP group. SAHA + DDP group could significantly reduce the colony forming ability of SAHA + DDP group. SAHA + DDP group could promote cleaved-Caspase-8 and cleaved-PARP Protein. The above results show that SAHA combined with cisplatin synergistic sensitization of osteosarcoma 143B cells can significantly inhibit its proliferation and migration and promote apoptosis.