Abstract
Evaluation of morphological changes in cells is an integral part of study on epithelial to mesenchymal transition (EMT), however, only a few papers reported the changes in quantitative parameters and no article compared different parameters for demanding better parameters. In the study, the purpose was to investigate suitable parameters for quantitative evaluation of EMT morphological changes. A549 human lung adenocarcinoma cell line was selected for the study. Some cells were stimulated by transforming growth factor-β1 (TGF-β1) for EMT, and other cells were as control without TGF-β1 stimulation. Subsequently, cells were placed in phase contrast microscope and three arbitrary fields were captured and saved with a personal computer. Using the tools of Photoshop software, some cells in an image were selected, segmented out and exchanged into unique hue, and other part in the image was shifted into another unique hue. The cells were calculated with 29 morphological parameters by Image Pro Plus software. A parameter between cells with or without TGF-β1 stimulation was compared statistically and nine parameters were significantly different between them. Receiver operating characteristic curve (ROC curve) of a parameter was described with SPSS software and F-test was used to compare two areas under the curves (AUCs) in Excel. Among them, roundness and radius ratio were the most AUCs and were significant higher than the other parameters. The results provided a new method with quantitative assessment of cell morphology during EMT, and found out two parameters, roundness and radius ratio, as suitable for quantification.
Highlights
Epithelial to mesenchymal transition (EMT) plays an important role in the metastasis of lung cancer, and has great therapeutic interest in the treatment of cancer [1,2], etc
Some quantitative parameters were used for the morphometry, including cellular size, roundness [12,13], but we did not find a report about suitable parameter choice on quantitative morphology of A549 cell in EMT
Images of cell shape in segment pre-and post (A) An image with points of grid stimulated by Transforming growth factor-β1 (TGF-β1). (B) An image with points of grid never stimulated by TGF-β1. (C) The image stimulated by TGF-β1 with segmented and sorted cells. (D) The image stimulated never by TGF-β1, with segmented and sorted cells
Summary
Epithelial to mesenchymal transition (EMT) plays an important role in the metastasis of lung cancer, and has great therapeutic interest in the treatment of cancer [1,2], etc. As a human lung adenocarcinoma cell, A549 cell has been applied widely in EMT study [3,4]. Transforming growth factor-β1 (TGF-β1), one of potent and well-studied cytokine, has been shown to alter the morphology of A549 cell line from cobblestone shape to elongated spindle shape, a fibroblastoid appearance [5,6,7,8]. The morphological change has been an important parameter in EMT studies, widely used for assessing anti-EMT effect of compounds, natural products [9,10], etc. In most studies, the morphological change was directly qualitative description [5,6,7,8,11], seldom evaluation by quantitative parameter. Some quantitative parameters were used for the morphometry, including cellular size, roundness [12,13], but we did not find a report about suitable parameter choice on quantitative morphology of A549 cell in EMT
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