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Pro-apoptotic effect of Δ2-TGZ in “claudin-1-low” triple-negative breast cancer cells: involvement of claudin-1

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Breast Cancer Research and Treatment Aims and scope Submit manuscript

An Erratum to this article was published on 31 July 2017

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Abstract

Purpose

40% of triple-negative breast cancer (TNBC) do not express claudin-1, a major constituent of tight junction. Patients with these “claudin-1-low” tumors present a higher relapse incidence. A major challenge in oncology is the development of innovative therapies for such poor prognosis tumors. In this context, we study the anticancer effects of ∆2-TGZ, a compound derived from troglitazone (TGZ), on cell models of these tumors.

Methods and results

In MDA-MB-231 and Hs578T “claudin-1-low” TNBC cells, Δ2-TGZ treatment induced claudin-1 protein expression and triggered apoptosis as measured by FACS analysis (annexin V/PI co-staining). Interestingly, in the non-tumorigenic human breast epithelial cell line MCF-10A, the basal level of claudin-1 was not modified following Δ2-TGZ treatment, which did not induce apoptosis. Furthermore, claudin-1-transfected MDA-MB-231 and Hs578T cells displayed a significant increase of cleaved PARP-1 and caspase 7, caspase 3/7 activities, and TUNEL staining. RNA interference was performed in order to inhibit Δ2-TGZ-induced claudin-1 expression in both the cells. In absence of claudin-1, a decrease of cleaved PARP-1 and caspase 7 and caspase 3/7 activities were observed in MDA-MB-231 but not in Hs578T cells.

Conclusion

Claudin-1 overexpression and Δ2-TGZ treatment are associated to apoptosis in MDA-MB-231 and Hs578T “claudin-1-low” TNBC. Moreover, in MDA-MB-231 cells, claudin-1 is involved in the pro-apoptotic effect of Δ2-TGZ. Our results suggest that claudin-1 re-expression could be an interesting therapeutic strategy for “claudin-1-low” TNBC.

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Change history

  • 31 July 2017

    An erratum to this article has been published.

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Acknowledgements

This work was supported by grants of the “Université de Lorraine,” the “Conseil Régional du Grand Est,” and the “Ligue Contre le Cancer.” Marine Geoffroy was recipient of a PhD grant of the «Ministère de l’Enseignement Supérieur et de la Recherche». We thank Christelle Thibault-Carpentier and Doulaye Dembele from the IGBMC GenomEast platform for the GEO deposit.

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Authors and Affiliations

  1. Université de Lorraine, CRAN, UMR 7039, 54506, Vandœuvre-lès-Nancy, France

    Marine Geoffroy, Alexandra Kleinclauss, Stéphanie Grandemange, Stéphane Flament, Isabelle Grillier-Vuissoz & Sandra Kuntz

  2. CNRS, CRAN, UMR 7039, 54506, Vandœuvre-lès-Nancy, France

    Marine Geoffroy, Alexandra Kleinclauss, Stéphanie Grandemange, Stéphane Flament, Isabelle Grillier-Vuissoz & Sandra Kuntz

  3. Université de Lorraine, SRSMC, UMR 7565, 54506, Vandœuvre-lès-Nancy, France

    Michel Boisbrun

  4. CNRS, SRSMC, UMR 7565, 54506, Vandœuvre-lès-Nancy, France

    Michel Boisbrun

  5. CNRS, FR3209 Biologie Moléculaire Cellulaire et Thérapeutique (BMCT), Plateforme d’Imagerie Cellulaire et Tissulaire PTIBC-IBISA, Biopôle de l’Université de Lorraine, Campus Biologie-Santé, 54506, Vandœuvre-lès-Nancy, France

    Sébastien Hupont

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Correspondence to Isabelle Grillier-Vuissoz.

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The original version of this article was revised: The given and family names of the authors were swapped. The corrected author group was updated in the article.

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Supplementary material 1 (PPTX 36 kb)

10549_2017_4378_MOESM2_ESM.pptx

Supplementary data. 1 Changes in expression level of genes related to cell adhesion and associated with cancer progression in Δ2-TGZ-treated MCF-7 cells after microarray analysis. Cells were treated for 12 h with DMSO or 25 µM of Δ2-TGZ Gene-expression microarray data were generated using GeneChip Human Gene 1.0 ST Array (containing 764,885 distinct probes including 28,869 well-annotated genes) as previously described [28]. Microarray data are available from Geo Data set (https://www.ncbi.nlm.nih.gov/geo/info/linking.html, accession number GSE99941). Supplementary material 2 (PPTX 39 kb)

10549_2017_4378_MOESM3_ESM.pptx

Supplementary data. 2 Claudin-1 and occludin co-localization in response to Δ2-TGZ in MDA-MB-231 and Hs578T cells. After treatment for 24 h with DMSO (Ctrl) or 20 µM of Δ2-TGZ, immunofluorescence staining for claudin-1 and occludin were performed in MDA-MB-231 and Hs578T cells. DRAQ5 staining was used for visualisation of the nuclei. a–b Cells were observed by confocal microscopy with magnification from ×400 for Hs578T to ×536 for MDA-MB-231 depending of digital confocal zooming combine with ×40 objective (NA 0.8). One confocal z section of cell of each condition is presented. Claudin-1 staining is upregulated and punctiform in both cell lines. Occludin staining is upregulated only in MDA-MB-231 cells and colocalized with claudin-1. Bar represents 50 µm. Supplementary material 3 (PPTX 2351 kb)

10549_2017_4378_MOESM4_ESM.pptx

Supplementary data. 3 Increase apoptosis in MDA-MB-231 cells overexpressing claudin-1 with Δ2-TGZ. MDA-MB-231 cells were transiently transfected with pcDNA3.1 or pcDNA3.1 containing a human claudin-1 expression vector (pcDNA3.1-CLDN1) and treated with 20 µM of Δ2-TGZ. After 48 h of transfection and 24 h of treatment, cells were subjected to western blot analysis. a Western blot analysis was performed with antibodies recognizing claudin-1 (CLDN1) cleaved PARP-1 and cleaved caspase 7. α-tubulin was used as loading control. b Relative protein level corresponds to claudin-1, cleaved PARP-1 (PARP) and cleaved caspase 7 band intensity value adjusted to α-tubulin for Δ2-pcDNA3.1 and Δ2-pcDNA3.1-CLDN1. The values represent the mean ± SEM 5 different experiments. Student-t test was used to determine significance difference from control cells, were *p < 0.05, **p < 0.01 and ***p < 0.001. Supplementary material 4 (PPTX 3130 kb)

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Geoffroy, M., Kleinclauss, A., Grandemange, S. et al. Pro-apoptotic effect of Δ2-TGZ in “claudin-1-low” triple-negative breast cancer cells: involvement of claudin-1. Breast Cancer Res Treat 165, 517–527 (2017). https://doi.org/10.1007/s10549-017-4378-2

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