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. 2018 Apr 3;8:5422. doi: 10.1038/s41598-018-23651-5

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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MSCs-derived B2M promotes epithelial-mesenchymal transition and enhances mobility of ESCC cells in vitro. (a) Morphological appearance of esophageal cancer cells treated with conditioned medium of MSC^NTC and MSC^shB2M suggested MSCs-derived B2M promotes EMT in vitro. In mesenchymal status, TE-1 present morphological changes from cobblestone-like to cord-like cells with sharp protuberance while Eca109 present incompact structure instead of clustered ‘islets’. Red arrows indicate representative morphological changes of tumor cells. (b,c) Immunofluorescence imaging revealed increased expression of N-cadherin (red) accompanied with decreased expression of E-cadherin (green) after treatments with MSC^NTC-CM but not with MSC^shB2M-CM in (b) TE-1 and (c) Eca109 cells. Nucleuses were immunostained with DAPI (blue). From left to right the images show the staining results for E-cadherin, N-cadherin, E-cadherin + DAPI, N-cadherin + DAPI, and Merge. (d,e) The qRT-PCR (upper panel) and western blot (lower panel) results showed elevated N-cadherin and vimentin with down-regulated E-cadherin in TE-1(d) and Eca109 (e) cancer cells after treated with MSC^NTC-CM in contrast to the MSC^shB2M-CM. (f) The number of the TE-1 cells in the migration (upper panel) and the invasion (lower panel) assays after treatment with MSC^NTC-CM was significantly increased compared with the negative control, while RNAi against B2M (MSC^shB2M) partially blocked this effect. (g) Similar migration and invasion results were observed in the Eca109 cells. Data is expressed as mean ± S.D. of three independent experiments. Error bars indicate S.D. (n = 3); *p < 0.05, **p < 0.01; CON: tumor cells untreated with conditioned medium; Scales bars: 100 μm.