Ibiyeye Kehinde Muibat, Nordin Norshariza, Ajat Mokrish, Zuki Abu Bakar Zakaria
Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Seri Kembangan, Malaysia.
Genetics and Regenerative Medicine Research Centre, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Seri Kembangan, Malaysia.
Front Oncol. 2019 Jul 3;9:599. doi: 10.3389/fonc.2019.00599. eCollection 2019.
Combination chemotherapy of anticancer drugs is extensively being researched since it could reduce multidrug resistance and side effects as a result of lower dosage of each drug. In this study, we evaluated the effects of doxorubicin-loaded (Dox-ACNP), thymoquinone-loaded (TQ-ACNP) and a combined doxorubicin/thymoquinone-loaded cockle shell-derived aragonite calcium carbonate nanoparticles (Dox/TQ-ACNP) on breast cancer cell line and compared with their free drugs counterpart. Cell viability using MTT assay, apoptosis with Annexin V-PI kit, morphological changes using contrast light microscope, scanning electron microscope and transmission electron microscope, cell cycle analysis, invasion assay, and scratch assay were carried out. The cell viability was evaluated in breast cancer cell line (MDA MB231), normal breast cells (MDF10A) and normal fibroblast (3T3). MDA MB231 IC dosages of drug-loaded nanoparticle were not toxic to the normal cells. The combination therapy showed enhanced apoptosis, reduction in cellular migration and invasion when compared to the single drug-loaded nanoparticle and the free drugs. Scanning electron microscope showed presence of cell shrinkage, cell membrane blebbing, while transmission electron microscope showed nuclear fragmentation, disruption of cell membrane, apoptotic bodies, and disruption of mitochondrial cistern. The results from this study showed that the combined drug-loaded cockle shell-derived aragonite calcium carbonate nanoparticles (Dox/TQ-ACNP) showed higher efficacy in breast cancer cells at lower dose of doxorubicin and thymoquinone.
由于联合化疗可通过降低每种药物的剂量来减少多药耐药性和副作用，因此抗癌药物的联合化疗正在广泛研究中。在本研究中，我们评估了负载阿霉素（Dox-ACNP）、负载百里醌（TQ-ACNP）以及负载阿霉素/百里醌的蛤壳衍生文石碳酸钙纳米颗粒（Dox/TQ-ACNP）对乳腺癌细胞系的影响，并与它们的游离药物对应物进行比较。使用MTT法检测细胞活力，用Annexin V-PI试剂盒检测细胞凋亡，用相差光显微镜、扫描电子显微镜和透射电子显微镜观察形态变化，进行细胞周期分析、侵袭实验和划痕实验。在乳腺癌细胞系（MDA MB231）、正常乳腺细胞（MDF10A）和正常成纤维细胞（3T3）中评估细胞活力。负载药物的纳米颗粒的MDA MB231 IC剂量对正常细胞无毒。与单一负载药物的纳米颗粒和游离药物相比，联合疗法显示出增强的细胞凋亡、细胞迁移和侵袭减少。扫描电子显微镜显示细胞收缩、细胞膜起泡，而透射电子显微镜显示核碎裂、细胞膜破坏、凋亡小体和线粒体池破坏。本研究结果表明，负载联合药物的蛤壳衍生文石碳酸钙纳米颗粒（Dox/TQ-ACNP）在较低剂量的阿霉素和百里醌下对乳腺癌细胞显示出更高的疗效。
