Department of Clinical Biochemistry, Tabriz University of Medical Sciences, Tabriz, Iran.
Molecular Medicine Research Center, Tabriz University of Medical Sciences, Iran.
Drug Res (Stuttg). 2022 Apr;72(4):220-225. doi: 10.1055/a-1775-7908. Epub 2022 Apr 6.
Osteosarcoma (OS) as the most frequent primary bone malignancy in children and adolescents has a short survival rate in advanced stages. Alternative herbal medicines with fewer side effects or the potency to protect common therapy's side effects can be helpful in combinational therapies. Herein, we aim to explore the effects of Thymoquinone (TQ) combined with Methotrexate (MTX) on Saos-2 cells apoptosis.
The effects of TQ and MTX alone or in combination on Saos-2 cell viability were measured by MTT assay. Real-time PCR was applied for the measurement of Bax, BCL-2, and caspase-9 mRNA expression. Apoptosis evaluation was conducted by flow cytometry.
TQ improves the cytotoxic effects of MTX on Saos-2 cells proliferation at lower doses. Indeed, the IC50 of MTX decreased from 26 μM to 15 μM when it combined with TQ. TQ and MTX can induce the expression level of pro-apoptotic factors, Bax and caspase-9 while inhibiting anti-apoptotic protein BCL-2. Moreover, the combination of TQ and MTX potentiates apoptosis to 73%, compared to either TQ (48%) or MTX (53%) treated cells.
The co-treatment of TQ and MTX is associated with the up-regulation of apoptotic factors and down-regulation of anti-apoptotic factors, conducting apoptosis aggravation and OS cell death.
骨肉瘤（OS）作为儿童和青少年中最常见的原发性骨恶性肿瘤，在晚期的生存率较低。具有较少副作用或能够保护常见治疗副作用的替代草药药物，可能有助于联合治疗。在此，我们旨在探讨姜黄素（TQ）联合甲氨蝶呤（MTX）对 Saos-2 细胞凋亡的影响。
通过 MTT 测定法测定 TQ 和 MTX 单独或联合对 Saos-2 细胞活力的影响。实时 PCR 用于测量 Bax、BCL-2 和 caspase-9 mRNA 的表达。通过流式细胞术评估细胞凋亡。
TQ 可在较低剂量下增强 MTX 对 Saos-2 细胞增殖的细胞毒性作用。实际上，当与 TQ 联合使用时，MTX 的 IC50 从 26 μM 降低到 15 μM。TQ 和 MTX 可诱导促凋亡因子 Bax 和 caspase-9 的表达水平，同时抑制抗凋亡蛋白 BCL-2。此外，与 TQ（48%）或 MTX（53%）处理的细胞相比，TQ 和 MTX 的联合处理可将细胞凋亡增加到 73%。
TQ 和 MTX 的联合治疗与上调促凋亡因子和下调抗凋亡因子有关，导致细胞凋亡加重和 OS 细胞死亡。
