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. 2020 Jul 11;39:131. doi: 10.1186/s13046-020-01610-1

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Silencing of HOTAIR facilitates chemosensitivity to 5-FU and apoptosis and reduces the proliferation of EC cells. Two shRNAs were delivered into TE-1/5-FU cells to specifically blunt HOTAIR (sh1-lncRNA HOTAIR and sh2-lncRNA HOTAIR group). Expression vectors containing the lncRNA HOTAIR were introduced into TE-1/5-FU cells (oe-lncRNA HOTAIR group). Scramble shRNA-treated cells and empty vectors-treated cells served as negative controls (sh-NC group and oe-NC group). (a) CCK-8 assay was performed to test the sensitivity of EC cell lines to 5-FU. (b) CCK-8 assay was performed to assess the surviving rate of TE-1/5-FU resistant strain in response to different concentrations of 5-FU. (c) CCK-8 assay was performed to assess the surviving rate of TE-1 cell line and TE-1/5-FU cell line at a 5-FU concentration of 5 μg/mL. (d) RT-qPCR was performed to determine the lncRNA HOTAIR expression in TE-1 cells and TE-1/5-FU cells, normalized to GAPDH. (e) Silencing and overexpression efficiency in TE-1/5-FU cells detected using RT-qPCR, normalized to GAPDH. (f) CCK-8 assay was performed to detect TE-1/5-FU cell viability following HOTAIR alternations. (g) CCK-8 assay was performed to measure IC₅₀ value in TE-1/5-FU cells following HOTAIR alternations. (h) EdU assay was performed to measure cell proliferation of TE-1/5-FU cells following HOTAIR alternations (× 400) (i) Flow cytometry was performed to assess apoptosis of TE-1/5-FU cells following HOTAIR alternations. (j) Flow cytometry was performed to assess cell cycle of TE-1/5-FU cells following HOTAIR alternations. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed using ANOVA followed by Tukey’s post hoc test among three or more groups. *p < 0.05 compared with TE-1/5-FU cells treated with either sh-NC or oe-NC group