Suitable parameter choice on quantitative morphology of A549 cell in epithelial-mesenchymal transition

Biosci Rep. 2015 Apr 22;35(3):e00202. doi: 10.1042/BSR20150070.

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.

Keywords: A549 cell; EMT; evaluation; morphological parameters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / drug therapy
  • Adenocarcinoma / pathology*
  • Adenocarcinoma of Lung
  • Area Under Curve
  • Cell Line, Tumor / drug effects
  • Epithelial-Mesenchymal Transition / physiology*
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / pathology*
  • Software
  • Transforming Growth Factor beta1 / pharmacology

Substances

  • Transforming Growth Factor beta1