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体外 MCF-7 超微结构变化用于转移性骨癌,通过 CaCO3/Dox 纳米晶体释放的线粒体细胞色素 C 诱导细胞凋亡。

In vitro ultrastructural changes of MCF-7 for metastasise bone cancer and induction of apoptosis via mitochondrial cytochrome C released by CaCO3/Dox nanocrystals.

机构信息

Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia (UPM), 43400 Serdang, Malaysia.

Faculty of Veterinary Medicine, Universiti Putra Malaysia (UPM), 43400 Serdang, Malaysia.

出版信息

Biomed Res Int. 2014;2014:391869. doi: 10.1155/2014/391869. Epub 2014 Jun 16.

Abstract

Bones are the most frequent site for breast cancer cells to settle and spread (metastasise); bone metastasis is considered to have a substantial impact on the quality of patients with common cancers. However, majority of breast cancers develop insensitivity to conventional chemotherapy which provides only palliation and can induce systemic side effects. In this study we evaluated the effect of free Dox and CaCO3/Dox nanocrystal on MCF-7 breast cancer using MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide), neural red, and lactate dehydrogenase colorimetric assays while DNA fragmentation and BrdU genotoxicity were also examined. Apoptogenic protein Bax, cytochrome C, and caspase-3 protein were analysed. Morphological changes of MCF-7 were determined using contrast light microscope and scanning and transmission electron microscope (SEM and TEM). The findings of the analysis revealed higher toxicity of CaCO3/Dox nanocrystal and effective cells killing compared to free Dox, morphological changes such as formation of apoptotic bodies, membrane blebbing, and absent of microvilli as indicated by the SEM analysis while TEM revealed the presence of chromatin condensation, chromosomal DNA fragmentation, cell shrinkage, and nuclear fragmentation. Results of TUNEL assay verified that most of the cells undergoes apoptosis by internucleosomal fragmentation of genomic DNA whereas the extent of apoptotic cells was calculated using the apoptotic index (AI). Therefore, the biobased calcium carbonate nanocrystals such as Dox carriers may serve as an alternative to conventional delivery system.

摘要

骨骼是乳腺癌细胞最常定居和扩散(转移)的部位;骨转移被认为对常见癌症患者的生活质量有重大影响。然而,大多数乳腺癌对常规化疗产生耐药性,常规化疗只能缓解症状,并可能引起全身性副作用。在这项研究中,我们使用 MTT(3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐)、神经红和乳酸脱氢酶比色法评估游离 Dox 和 CaCO3/Dox 纳米晶体对 MCF-7 乳腺癌的影响,同时还检查了 DNA 片段化和 BrdU 遗传毒性。分析了凋亡蛋白 Bax、细胞色素 C 和 caspase-3 蛋白。使用相差显微镜和扫描电子显微镜(SEM)和透射电子显微镜(TEM)观察 MCF-7 的形态变化。分析结果显示,与游离 Dox 相比,CaCO3/Dox 纳米晶体具有更高的毒性和有效的细胞杀伤作用,SEM 分析表明,形态学变化包括凋亡小体的形成、细胞膜起泡和微绒毛缺失,而 TEM 显示染色质浓缩、染色体 DNA 片段化、细胞收缩和核碎裂。TUNEL 检测结果证实,大多数细胞通过基因组 DNA 的核小体间断裂发生凋亡,而通过凋亡指数(AI)计算凋亡细胞的程度。因此,基于生物的碳酸钙纳米晶体等 Dox 载体可以作为传统递送系统的替代物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/edf6/4084513/ce4e90cab6ac/BMRI2014-391869.001.jpg

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