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. 2020 Jan 28;10:1290. doi: 10.1038/s41598-020-57940-9

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© The Author(s) 2020

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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Cucurbitacin B and I inhibits spheroid formation. (A) HCT116 and SW480 cells were grown in specific spheroid media in low adherent plates and treated with increasing concentrations of C-B and C-I (2.5, 5 and 10 µM). After 5 days, the colonosphere were photographed. The primary spheroids were collected and separated into single cells and replated. The C-B and C-I treatment significantly were inhibited in both primary and secondary colonosphere formation. (B) Quantification of spheroid number from spheroid formation assay. The C-B and C-I treatment significantly were inhibited in the number of colonosphere formation (*p < 0.01). (C) Western blot analyses of cells lysates after C-B and C-I treatment showed a significant reduction in cancer stem cell marker DCLK1 (+19 and −19 isoform), DCLK1 (−19 isoform), CD44, and LGR5 protein levels in both HCT116 and SW480 cells.