Conditioned media from activated healthy donor PBMC induces EMT in IBC.

<p>(a) Morphological changes visualized by bright field in IBC cell lines consistent with stress and EMT were observed following 48-hour incubation with CM. (b) Immunohistochemical stains of paraffin cell blocks: the percentage of positive cells is listed in each image and shown in the bar graph at right. Pancytokeratin expression is shown for both percent of cells with membrane localization (top number) and cytoplasmic localization (bottom number). MCF-7 control cells show a characteristic epithelial phenotype with high E-cadherin, low vimentin, low keratin 5/6 expression and strong membrane and cytoplasmic localization of cytokeratins, MDA-231 control cells are mostly mesenchymal with low E-cadherin, high vimentin and decreased cytokeratins. Following exposure to TCR-CM, SUM149 cells show increased expression of E-cadherin, vimentin, keratin 5/6 staining and decreased pan cytokeratin staining. (c) Expression levels of EMT-related transcription factors SNAIL, ZEB1 and TG2 were quantified by Taq-Man qRT-PCR. TCR-CM and to a lesser extent LPS-CM, induced large increases in ZEB1 and TG2.