MORPHOLOGICAL ANALYSIS OF COLON CANCER CELL TREATED WITH AEE788, A DUAL TYROSINE KINASE INHIBITOR AND/OR CELEXOXIB, A COX-2 SPECIFIC INHIBITOR- IMAGE PROCESSING APPROACH

摘要

Evaluation of morphological changes in cancer cell provoked by external stimuli is the focus of current cancer therapeutic study. Present investigation is aimed to evaluate efficacy of the drug by quantitative analysis of morphological features of the colon cancer cells. Standard optical microscopy used for morphological analysis has limitation in resolution (>200nm) due to diffraction limit which control fine microscopic structure associated with cancer cell. But, disadvantages associated with phase contrast microscopy are faint images, inaccurate detection and diffraction limit etc., Consequently, observation of anticancer agent mediated morphological changes in cancer cell can be of difficult. In this study, we analysed morphology of AEE788 and/or celecoxib treated HCT 15 cell using confocal laser microscopy. Nevertheless, this observation is solely qualitative and often leads to considerable variability. To improve the reliability of confocal microscopic analysis by minimizing the subjectivity, it is important to analyze the morphological features from the image in a quantitative way using digital image processing techniques. Hence, the cytoplasm and nucleus of cell are extracted using color deconvolution followed by Otsu's thresholding. Then the features like area, perimeter, eccentricity, compactness and Fractal Dimension (FD) are quantitatively estimated to mathematically describe the cellular morphology. The results show the reduction in area, perimeter, compactness and eccentricity of drug-treated cells compared to untreated control HCT 15. In contrast, the FD values of these drug-treated cells are increasing as compared to control HCT 15, which indicate the membrane damage and blebbing effect of the drugs. Moreover, this study provides quantitative information about morphological changes in colon cancer cell line which in turn indicates the efficacy of AEE788 and/or Celexcoxib. In conclusion, this quantitative analysis can be used as a preliminary experiment along with other established techniques in evaluating the efficacy of anticancer agents.