Colcemid duration effects on extrachromosomal DNA analysis in breast cancer cell line

IF 2.7 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2025-05-05 DOI:10.1016/j.tice.2025.102952

Shadira Anindieta Irdianto , Fadhillah Fadhillah , Retno Lestari , Fadilah Fadilah , Astari Dwiranti , Anom Bowolaksono

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Abstract

Extrachromosomal DNA (ecDNA) is a circular form of genetic material that exists outside the chromosomes and plays a significant role in tumour heterogeneity and drug resistance. In this preliminary study, we aimed to optimize the visualization of ecDNA by investigating the effects of varying colcemid treatment durations on metaphase arrest in MDA-MB-231 breast cancer cells. We tested four durations of colcemid treatment: 20 hours, 3 hours, 1 hour, and 30 minutes. Using light microscopy, confocal microscopy, and scanning electron microscopy, we analyzed the morphology and presence of ecDNA. Our results demonstrated that 20-hour colcemid treatment yielded the most well-spread metaphase cells, enabling the optimal observation of ecDNA. These structures appeared as ring-like formations with lower DAPI intensity than chromosomes and were often found in pairs. While our findings contribute valuable insights into the visualization of ecDNA, they also serve as a foundation for future research.

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秋氨酸持续时间对乳腺癌细胞系染色体外DNA分析的影响

染色体外DNA （ecDNA）是一种存在于染色体外的圆形遗传物质，在肿瘤异质性和耐药性中起着重要作用。在这项初步研究中，我们旨在通过研究不同秋碱治疗时间对MDA-MB-231乳腺癌细胞中期阻滞的影响来优化ecDNA的可视化。我们测试了四种持续时间：20 小时、3 小时、1 小时和30 分钟。使用光学显微镜、共聚焦显微镜和扫描电镜，我们分析了ecDNA的形态和存在。我们的研究结果表明，20小时的秋碱处理产生了最广泛的中期细胞，使ecDNA的观察效果最佳。这些结构呈环状结构，DAPI强度低于染色体，通常成对出现。虽然我们的发现为ecDNA的可视化提供了有价值的见解，但它们也为未来的研究奠定了基础。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.