Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, Abha, Saudi Arabia.
Central Research Laboratory, College of Applied Medical Sciences, King Khalid University, Abha, Saudi Arabia.
Arch Pharm Res. 2020 Jun;43(6):655-665. doi: 10.1007/s12272-020-01241-2. Epub 2020 Jun 25.
In this study, we report the combination of indirubin-3-monoxime (I3M) and thymoquinone (Tq) to have excellent therapeutic efficacy in models of Lung cancer (LC). Preliminary screening was done with A549 cells. Cell cycle, apoptosis and NFκB phosphorylation were determined by flow cytometry, while apoptotic proteins, Akt and mTOR were assessed by western blotting. Mouse xenograft model was used to assess the therapeutic efficacy in-vivo. Synergistic reduction in cell viability was observed with I3M + Tq combinations, which were non-toxic to normal HFL-1 cells. Cell cycle analysis indicated G phase reduction with subsequent accumulation of sub G contents. Annexin V assay revealed higher apoptotic cells with combinations compared to individual treatments with a decrease in Bcl-2/Bax ratio. The combinations exhibited anti-metastasis activity in cell migration in the scratch, scatter and tumour cell migration assays and effectively reduced the tumour growth in mouse xenograft model. Expression levels of p-AKT, p-mTOR, Caspase-3, p-53 and NFκB were significantly reduced in the combination treated mice compared to individual treatments. Results of current study demonstrate clear efficacy of I3M + Tq combinations in LC models mediated by suppressing Akt/mTOR/NFκB signalling. Further research is recommended to transform these findings into novel therapeutic combinations against LC.
在这项研究中，我们报告了靛玉红-3-单肟（I3M）和百里醌（Tq）联合应用在肺癌（LC）模型中具有优异的治疗效果。初步筛选采用 A549 细胞进行。通过流式细胞术测定细胞周期、凋亡和 NFκB 磷酸化，通过 Western blot 测定凋亡蛋白、Akt 和 mTOR。使用小鼠异种移植模型评估体内治疗效果。I3M+Tq 联合用药可显著降低细胞活力，且对正常 HFL-1 细胞无毒。细胞周期分析表明 G 期减少，随后亚 G 含量增加。与单独治疗相比，组合治疗后的 Annexin V 测定显示凋亡细胞增多，Bcl-2/Bax 比值降低。在划痕、散射和肿瘤细胞迁移测定中，联合用药具有抗转移活性，并能有效抑制小鼠异种移植模型中的肿瘤生长。与单独治疗相比，在联合治疗的小鼠中，p-AKT、p-mTOR、Caspase-3、p-53 和 NFκB 的表达水平显著降低。本研究结果表明，I3M+Tq 联合用药在 LC 模型中具有明显的疗效，其作用机制可能与抑制 Akt/mTOR/NFκB 信号通路有关。建议进一步研究将这些发现转化为针对 LC 的新型治疗联合用药。
