Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Selangor 43400, Malaysia.
Department of Veterinary Preclinical Sciences, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor 43400, Malaysia.
Int J Mol Sci. 2020 Mar 10;21(5):1900. doi: 10.3390/ijms21051900.
Cancer stem cells CSCs (tumour-initiating cells) are responsible for cancer metastasis and recurrence associated with resistance to conventional chemotherapy. This study generated MBA MD231 3D cancer stem cells enriched spheroids in serum-free conditions and evaluated the influence of combined doxorubicin/thymoquinone-loaded cockle-shell-derived aragonite calcium carbonate nanoparticles. Single loaded drugs and free drugs were also evaluated. WST assay, sphere forming assay, ALDH activity analysis, Surface marker of CD44 and CD24 expression, apoptosis with Annexin V-PI kit, cell cycle analysis, morphological changes using a phase contrast light microscope, scanning electron microscopy, invasion assay and migration assay were carried out; The combination therapy showed enhanced apoptosis, reduction in ALDH activity and expression of CD44 and CD24 surface maker, reduction in cellular migration and invasion, inhibition of 3D sphere formation when compared to the free drugs and the single drug-loaded nanoparticle. Scanning electron microscopy showed poor spheroid formation, cell membrane blebbing, presence of cell shrinkage, distortion in the spheroid architecture; and the results from this study showed that combined drug-loaded cockle-shell-derived aragonite calcium carbonate nanoparticles can efficiently destroy the breast CSCs compared to single drug-loaded nanoparticle and a simple mixture of doxorubicin and thymoquinone.
癌症干细胞（CSCs，肿瘤起始细胞）是导致癌症转移和复发的原因，与对常规化疗的耐药性有关。本研究在无血清条件下生成 MBA MD231 三维癌症干细胞富集球体，并评估了载多柔比星/百里醌的贝壳衍生文石碳酸钙纳米颗粒的联合治疗效果。还评估了单载药物和游离药物的影响。进行了 WST 检测、球体形成检测、ALDH 活性分析、CD44 和 CD24 表达的表面标志物分析、用 Annexin V-PI 试剂盒进行细胞凋亡分析、细胞周期分析、相差光显微镜下的形态变化、扫描电子显微镜、侵袭和迁移实验；与游离药物和单载药纳米颗粒相比，联合治疗显示出增强的细胞凋亡、ALDH 活性和 CD44 和 CD24 表面标志物表达的降低、细胞迁移和侵袭的减少、三维球体形成的抑制。扫描电子显微镜显示球体形成不良、细胞膜起泡、细胞收缩、球体结构扭曲；本研究结果表明，与单载药纳米颗粒和多柔比星与百里醌的简单混合物相比，联合载药贝壳衍生文石碳酸钙纳米颗粒可以有效地破坏乳腺癌 CSCs。
