Multiplex immunohistochemical characterization of Aurora A and Aurora B in breast, ovarian, non-small cell lung, and colon cancer Free

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Aurora A and Aurora B are frequently over-expressed in a variety of cancers, and their inhibition has been demonstrated to cause selective apoptosis of tumor cells. Many investigation drugs target one or both of these molecules, and clinical trials are in progress to determine efficacy. The present study used an Aurora A / Aurora B / ki67 multiplex immunohistochemistry (IHC) assay to study expression of the Auroras in breast, ovarian, non-small cell lung, and colon cancer in addition to normal tissues. Unsynchronized and nocodazole-treated HeLa cells were analyzed to demonstrate assay specificity, and increased expression of Aurora A and Aurora B was observed in nocodazole treated cells. Stained samples of cancer and normal tissue were imaged using multispectral imaging, and quantitative grayscale images for each analyte were created by spectral unmixing. The quantitative grayscale images were analyzed using image analysis software to determine the overall expression levels of Aurora A and Aurora B expression and the relationship between the two was evaluated using a 2-D scatterplot. The relationship between Aurora staining and ki67 was also evaluated. Evaluation and classification of cancers by multiplex Aurora analysis may provide critical insight into factors that predict response to Aurora kinase inhibitors in the clinic.