Department of Pharmacology, SVKM's Dr. Bhanuben Nanavati College of Pharmacy, Mumbai 400 056, Maharashtra, India.
Department of Pharmacology, SVKM's Dr. Bhanuben Nanavati College of Pharmacy, Mumbai 400 056, Maharashtra, India.
Biomed Pharmacother. 2018 Nov;107:1037-1045. doi: 10.1016/j.biopha.2018.08.095. Epub 2018 Aug 24.
Heat shock protein 70 (Hsp70) and heat shock protein 90 (Hsp90) chaperones are indispensable to lung cancer cells for their survival and proliferation. In this study we evaluated and compared anticancer potential of methylene blue (MB) as an Hsp70 inhibitor, novobiocin (NB) a well-known Hsp90 inhibitor and their combination.
In vitro evaluation was done by cell viability assays, fluorescent staining, and flow cytometry analysis using A549 non-small cell lung cancer cells. In vivo anticancer activity was investigated by evaluating oxidative stress, tumor biomarkers, weight, lung microarchitecture, and Hsp70 and Hsp90 inhibitions via immunoblotting in benzo[a]pyrene induced lung carcinogenesis mice model.
Using A549 NSCLC cells, we found MB demonstrated lower cell viability versus NB. Together, MB + NB resulted in further decrease in cell viability. SRB assay revealed significantly superior and similar potency for MB versus NB and MB + NB (1:1) versus MB, respectively. Fluorescent staining and flow cytometry analysis displayed early apoptosis by MB (11.4%); early and late apoptosis by MB + NB (13.8%). In vivo, MB significantly inhibited Hsp70. Furthermore, MB significantly alleviated tumor biomarkers (ADA and LDH) and improved lung histopathological features more than NB. Additionally, MB significantly improved SOD, not more than MB + NB or NB and improved LPO.
MB demonstrated potent anticancer activity in vitro and in vivo via inhibition of Hsp70 in benzo[a]pyrene induced lung carcinogenesis in mice.
热休克蛋白 70（Hsp70）和热休克蛋白 90（Hsp90）伴侣蛋白对于肺癌细胞的存活和增殖是必不可少的。在这项研究中，我们评估并比较了亚甲蓝（MB）作为 Hsp70 抑制剂、新生霉素（NB）作为一种著名的 Hsp90 抑制剂及其组合的抗癌潜力。
通过细胞活力测定、荧光染色和流式细胞术分析，使用 A549 非小细胞肺癌细胞进行体外评估。通过评估氧化应激、肿瘤生物标志物、体重、肺微结构以及通过免疫印迹法评估 Hsp70 和 Hsp90 抑制作用，研究苯并[a]芘诱导的肺癌发生小鼠模型中的体内抗癌活性。
使用 A549 NSCLC 细胞，我们发现 MB 与 NB 相比表现出更低的细胞活力。MB+NB 联合使用导致细胞活力进一步降低。SRB 测定显示 MB 对细胞活力的抑制作用显著优于 NB（1:1）和 MB+NB（1:1），与 NB 相比分别具有相似的效力。荧光染色和流式细胞术分析显示 MB 诱导早期凋亡（11.4%）；MB+NB 诱导早期和晚期凋亡（13.8%）。体内，MB 显著抑制 Hsp70。此外，MB 比 NB 更显著地减轻肿瘤生物标志物（ADA 和 LDH）并改善肺组织病理学特征。此外，MB 显著提高 SOD，不优于 MB+NB 或 NB，并且提高 LPO。
MB 通过抑制苯并[a]芘诱导的肺癌发生小鼠中的 Hsp70，在体外和体内均表现出强大的抗癌活性。
