Biochemistry Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
Biochemistry Department, Faculty of Pharmacy, Ahram Canadian University, Cairo, Egypt.
Chem Biol Interact. 2015 May 25;233:147-56. doi: 10.1016/j.cbi.2015.03.028. Epub 2015 Apr 8.
Imatinib mesylate (IM), a tyrosine kinase inhibitor, is used as targeted cancer therapy. However, mono-targeting by IM does not always achieve full tumor eradication and thus it is recommended to combine IM with other anticancer agents. Clotrimazole (CLT) is an antifungal azole derivative with promising anticancer effects due to inhibiting the activity of glycolytic enzymes. The present study aimed to evaluate the effect of combining CLT with IM on breast cancer cell line in an attempt to establish effective new combination. T47D human breast cancer cell line was treated with different concentrations of IM and/or CLT for 48 h. IM-CLT interaction was determined by isobologram equation and combination index. Cell viability was confirmed by measuring LDH activity. As indicators of glycolysis inhibition, the expression of hexokinase-2 (HK-2) and 6-phosphofructo-1-kinase (PFK-1) plus the activity of intracellular lactate dehydrogenase (LDH) and pyruvate kinase (PK) were determined. In addition, glucose consumption and adenosine triphosphate (ATP) production were measured. Moreover, nitric oxide (NO), vascular endothelial growth factor (VEGF) and hypoxia inducible factor-α (HIF-α) were also determined as they are modulators for glycolysis. This study demonstrated that IM or CLT synergistically inhibited cell growth in T47D as shown by combination and dose reduction indices. The combination of 15 μM IM and 20 μM CLT significantly decreased glucose consumption, activity of both PK and intracellular LDH, while increased leaked LDH, VEGF and NO in the medium compared to each drug alone. Furthermore the combination decreased gene expression of HK-2, PFK-1 and ATP content compared to the control. In conclusion, the synergistic effect of CLT on IM cytotoxicity in T47D cell line maybe mediated through inhibition of glycolysis and increasing both NO and VEGF. Further studies are required to confirm the efficiency and safety of this combination.
甲磺酸伊马替尼（IM）是一种酪氨酸激酶抑制剂，用于癌症的靶向治疗。然而，IM 的单靶点治疗并不总能完全清除肿瘤，因此建议将 IM 与其他抗癌药物联合使用。克霉唑（CLT）是一种抗真菌唑类衍生物，由于抑制糖酵解酶的活性，具有有前途的抗癌作用。本研究旨在评估 CLT 与 IM 联合应用于乳腺癌细胞系的效果，试图建立有效的新联合方案。用不同浓度的 IM 和/或 CLT 处理 T47D 人乳腺癌细胞系 48 小时。通过等效应线方程和合并指数确定 IM-CLT 相互作用。通过测量 LDH 活性来确认细胞活力。作为糖酵解抑制的指标，测定己糖激酶-2（HK-2）和 6-磷酸果糖-1-激酶（PFK-1）的表达以及细胞内乳酸脱氢酶（LDH）和丙酮酸激酶（PK）的活性。此外，还测量了葡萄糖消耗和三磷酸腺苷（ATP）的产生。此外，还测定了一氧化氮（NO）、血管内皮生长因子（VEGF）和缺氧诱导因子-α（HIF-α），因为它们是糖酵解的调节剂。本研究表明，IM 或 CLT 协同抑制 T47D 细胞生长，表现为合并和剂量减少指数。与单独使用每种药物相比，15 μM IM 和 20 μM CLT 的组合显著降低了葡萄糖消耗、PK 和细胞内 LDH 的活性，同时增加了培养基中漏出的 LDH、VEGF 和 NO。此外，与对照组相比，该组合降低了 HK-2、PFK-1 的基因表达和 ATP 含量。总之，CLT 对 T47D 细胞系中 IM 细胞毒性的协同作用可能是通过抑制糖酵解和增加 NO 和 VEGF 来介导的。需要进一步研究来确认这种组合的有效性和安全性。
