基于激光干涉显微镜相衬成像分析MCF-7肿瘤细胞在凋亡过程中的释放

A. Ignatova, A. Nikityuk, Y. Bayandin, V. Grishko, O. Naimark
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引用次数: 0

摘要

细胞表面形貌的数据使预测其行为成为可能,并且是评估其是否处于凋亡状态的指标,即由于细胞内反应的激活而处于死亡状态;细胞的这种状态既发生在正常发育过程中,也作为病理过程的结果。目前还没有一种通用的方法来记录细胞的松弛状态及其其他机械生物学参数,因此激光干涉显微镜很有希望用于这一目的。本研究的目的是通过分析激光干涉显微镜获得的相对比(phase)图像来评估乳腺癌细胞凋亡变化过程中肿瘤细胞表面的状态。本研究的对象是在阿霉素作用下处于天然状态和诱导凋亡状态的MCF-7细胞系(人乳腺上皮样腺癌)细胞。细胞表面的分析包括解释细胞的光学厚度及其形态的数据,通过构建等值线。通过对细胞相图的分析,确定了细胞的一般形态计量参数。使用imagj - fiji软件对光学数据进行可视化及其解释。根据Wilcoxnon-Mann-Whitney检验,所研究的样品在最大直径、周长和球度系数上存在显著差异。在等线图像分析的基础上,构建了细胞在自然状态和凋亡状态下的相态图。从图中可以看出,在相同高度水平下,凋亡细胞的横截面积通常比天然细胞的横截面积小得多。所提出的相图框架内的细胞特征与使用经典显微镜方法记录的形态学细胞凋亡信号一致。细胞状态图的构建,根据其救济的参数,有希望评估细胞的行为,预测它,以及分析药物和治疗方法对病理细胞的影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Analysis of the relief of MCF-7 tumor cells during apoptisis on the basis of phase-contrast images obtained by laser interference microscopy
Data on the surface topography of the cell makes it possible to predict its behavior and is an indicator to assess whether it is in a state of apoptosis, that is, in a state of death due to the activation of intracellular reactions; this state of a cell occurs both during normal development and as a result of a pathological process. A universal method for recording the state of the relief of the cell and its other mechanobiological parameters has not been developed, and therefore laser interference microscopy is promising for this purpose. The aim of this work is to evaluate the state of the surface of tumor cells during the apoptotic changes in breast cancer cells by analyzing the phase contrast (phase) images obtained by laser interference microscopy. The object of the study is the cells of the MCF-7 line (epithelial-like adenocarcinoma of the human mammary gland) in a native state and in a state of induced apoptosis under the action of doxorubicin. The analysis of the cell surface involves interpretation of data on the optical thickness of cells and their morphology through the construction of isolines. Based on the analysis of phase images of cells, their general morphometric parameters were determined. Visualization of optical data and their interpretation was performed using the ImageJ–Fiji software. According to the Wilcoxnon–Mann–Whitney test, the samples under study are significantly different in maximum diameter, perimeter, and sphericity coefficient. Based on the analysis of isolinear images, a diagram of the phase state of cells in a native state and in a state of apoptosis was constructed. According to the diagram, the cross-sectional area of apoptotic cells is generally much smaller than the cross-sectional area of native cells at the same height levels. The characteristics of cells within the framework of the proposed phase diagram are consistent with the morphological cell apoptosis signs, which are recorded using classical microscopic methods. The construction of the diagrams of the state of cells according to the parameters of their relief holds promise for assessing the cellular behavior, predicting it, as well as for analyzing the effect of drugs and therapeutic methods on pathological cells.
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