Clotam enhances anti-proliferative effect of vincristine in Ewing sarcoma cells.
Clotam enhances anti-proliferative effect of vincristine in Ewing sarcoma cells.
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Clotam 增强长春新碱对尤文肉瘤细胞的抗增殖作用。
DOI：
10.1007/s10495-018-1508-1
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Basha,Riyaz
中科院分区：
文献类型：
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作者：
Shelake,Sagar;Sankpal,UmeshT;Eslin,Don;Bowman,WPaul;Simecka,JerryW;Raut,Sangram;Ray,Anish;Basha,Riyaz
在过去的20年里，尤文肉瘤（ES）的治疗策略，特别是复发患者的治疗策略，使生存率略有提高。联合治疗方法是克服转移性ES耐药性的一种替代方法。本研究评价了Clotam（托芬那酸或TA）（一种小分子和特异性蛋白1（Sp1）和生存素抑制剂）对ES细胞系化疗药物长春新碱（VCR）敏感性的影响。用TA或VCR或TA + VCR（组合）处理ES细胞（CHLA-9和TC-32），并在24/48/72 h后评估细胞活力。应用碘化丙啶、细胞周期分析和Annexin V流式细胞仪分别检测TA、VCR或TA + VCR处理对细胞周期阻滞和凋亡的影响。测定细胞凋亡标志物caspase 3/7（活性水平）和裂解PARP（蛋白表达）。心肌细胞H9 C2用作非恶性细胞。然而，所有处理引起细胞活力的时间和剂量依赖性抑制，有趣的是，组合处理引起显著更高的响应（~ 80%抑制，p< 0.05）。细胞活力抑制伴随着Sp1和Survivin的抑制。TA + VCR处理显著（p< 0.05）增加caspase 3/7活性，这与上调的c-PARP水平和Annexin V染色密切相关。细胞周期阻滞在G 0/G1（TA）或G2/M（VCR和TA + VCR）。所有处理均未引起H9 C2细胞的细胞毒性。以上结果提示TA可增强VCR对ES细胞的抗癌活性。因此，可进一步检测TA + VCR组合，以开发治疗ES的安全/有效治疗策略。
Current therapeutic strategies used in Ewing sarcoma (ES) especially for relapsed patients have resulted in modest improvements in survival over the past 20 years. Combination therapeutic approach presents as an alternative to overcoming drug resistance in metastatic ES. This study evaluated the effect of Clotam (tolfenamic acid or TA), a small molecule and inhibitor of Specificity protein1 (Sp1) and survivin for sensitizing ES cell lines to chemotherapeutic agent, vincristine (VCR). ES cells (CHLA-9 and TC-32) were treated with TA or VCR or TA + VCR (combination), and cell viability was assessed after 24/48/72 h. Effect of TA or VCR or TA + VCR treatment on cell cycle arrest and apoptosis were evaluated using propidium iodide, cell cycle assay and Annexin V flow cytometry respectively. The apoptosis markers, caspase 3/7 (activity levels) and cleaved-PARP (protein expression) were measured. Cardiomyocytes, H9C2 were used as non-malignant cells. While, all treatments caused time- and dose-dependent inhibition of cell viability, interestingly, combination treatment caused significantly higher response (~ 80% inhibition,p< 0.05). Cell viability inhibition was accompanied by inhibition of Sp1 and Survivin. TA + VCR treatment significantly (p< 0.05) increased caspase 3/7 activity which strongly correlated with upregulated c-PARP level and Annexin V staining. Cell cycle arrest was observed at G0/G1 (TA) or G2/M (VCR and TA + VCR). All treatments did not cause cytotoxicity in H9C2 cells. These results suggest that TA could enhance the anti-cancer activity of VCR in ES cells. Therefore, TA + VCR combination could be further tested to develop as safe/effective therapeutic strategy for treating ES.