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. 2023 Oct 9;21:702. doi: 10.1186/s12967-023-04519-3

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© The Author(s) 2023

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 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. 4 — Gemcitabine stimulated human pancreatic stellate cells further enhances the gemcitabine resistance of MAGEA2-expressing tumor cells via a paracrine signalling. A Schematic diagram of the experimental design to test the effect of conditioned medium (CM) from placebo/Gem treated PSC in MIA PaCa-2 derived VA and MAGEA2 clones. B MIA PaCa-2 derived VA and MAGEA2 C10 clones were exposed with the CM of placebo or Gem treated PSC -/ + Gem for up to 5 days. C MAGEA2- or VA-expressing MIA PaCa-2 spheroids were cultured with either the CM of placebo or Gem treated PSC together with placebo or Gem for up to 5 days. D The CM harvested from placebo/Gem treated PSCs were first incubated with or without trypsin for 24 h at 37 °C, which were then added to MIA PaCa-2 derived MAGEA2 and VA clones for up to 120 h. E Quantitation of the fold difference in cytokine expression between placebo and Gem treated PSCs (n = 4 dots from 2 independent experiments). Representative pictures of the cytokine arrays derived from placebo or Gem treated PSCs. Arrows indicate the cytokines with significant change. F, G Analysis of TCGA database revealed that high expression of GDF15 and MAGEA was associated with poor overall survival and enhanced lymph node metastases in PDAC patients (n = 89 patients, TCGA dataset). H Receptor tyrosine array. Quantification of fold difference in p-RET level between MIA PaCa-2 derived VA and MAGEA2 clones after exposure with the CM of Gem stimulated PSCs. I Western blot analysis of MIA PaCa-2 derived VA and A2 C10 clones after treated with the CM of Gem stimulated PSCs. J Western blot analysis of PSCs transfected with non-silencing control (siNSC) or GDF15 targeting siRNA molecules for 48 h. K Line graph shows the cell growth of MIA PaCa2-MAGEA2 and VA clones after exposed with CM from siGDF15 depleted PSCs in the presence/absence of Gem over time (n = 3 independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. B–D, K Two-way ANOVA. E, G, H Student’s t test. F Log-rank (Mantel-Cox) test. Scale bars in (C) represent 200 μm