Hou Mengyi, Huang Zhenglan, Chen Sicheng, Wang Hao, Feng Tianyu, Yan Shujuan, Su Yuxi, Zuo Guowei
Key Laboratory of Diagnostic Medicine Designated by The Chinese Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Department of Clinical Medicine, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China.
Oncol Lett. 2018 Oct;16(4):4663-4670. doi: 10.3892/ol.2018.9224. Epub 2018 Jul 27.
Cisplatin, as a first-line chemotherapy drug, has been widely applied for therapy of osteosarcoma. However, its application is limited by drug resistance and serious side effects, including nephrotoxicity and ototoxicity. Suberoylanilide hydroxamic acid (SAHA) is a newly developed histone deacetylase (HDAC) inhibitor, which is the first Food and Drug Administration-approved HDAC inhibitor for the treatment of cutaneous manifestations of T-cell lymphoma. However, SAHA as a monotherapy was revealed to be limited, particularly in solid tumors. In the present study, 143B osteosarcoma cells were treated with multiple concentrations of SAHA or cisplatin, either alone or combined. The morphological characteristics of the treated cells were observed using an inverted microscope. The cytotoxicity effects of the combination of SAHA and cisplatin on 143B cells were analyzed by MTT assay, colony formation assay, wound healing cell migration assay, cell apoptosis assay and cell cycle analysis. Western blot analysis was performed to detect the protein expression levels of B cell lymphoma-2 (Bcl-2)-associated X protein (Bax), Bcl-2, cleaved-caspase-3, cleaved-caspase-8 and cleaved-poly (ADP-ribose) polymerase (PARP). The experimental data indicated that the inhibition of cell proliferation in the combination group was significantly increased compared with that in single drug groups. Expression levels of pro-apoptotic protein were upregulated, whereas anti-apoptotic Bcl-2 was downregulated significantly in 143B cells following SAHA/cisplatin treatment. Taken together, the results revealed that the combination of SAHA and cisplatin inhibited the proliferation of 143B cells and induced their apoptosis synergistically, and this effectiveness may be mediated by caspase activation.
顺铂作为一线化疗药物,已被广泛应用于骨肉瘤的治疗。然而,其应用受到耐药性和严重副作用的限制,包括肾毒性和耳毒性。伏立诺他是一种新开发的组蛋白去乙酰化酶(HDAC)抑制剂,是首个获得美国食品药品监督管理局批准用于治疗T细胞淋巴瘤皮肤表现的HDAC抑制剂。然而,伏立诺他作为单一疗法显示出局限性,尤其是在实体瘤中。在本研究中,用多种浓度的伏立诺他或顺铂单独或联合处理143B骨肉瘤细胞。使用倒置显微镜观察处理后细胞的形态特征。通过MTT法、集落形成试验、伤口愈合细胞迁移试验、细胞凋亡试验和细胞周期分析,分析伏立诺他和顺铂联合对143B细胞的细胞毒性作用。进行蛋白质免疫印迹分析以检测B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)、Bcl-2、裂解的半胱天冬酶-3、裂解的半胱天冬酶-8和裂解的聚(ADP-核糖)聚合酶(PARP)的蛋白质表达水平。实验数据表明,联合组细胞增殖的抑制作用比单药组显著增强。SAHA/顺铂处理后,143B细胞中促凋亡蛋白的表达水平上调,而抗凋亡蛋白Bcl-2显著下调。综上所述,结果表明,伏立诺他和顺铂联合抑制了143B细胞的增殖并协同诱导其凋亡,这种有效性可能是由半胱天冬酶激活介导的。