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Twist1-induced activation of human fibroblasts promotes matrix stiffness by upregulating palladin and collagen α1(VI)

Oncogene volume 35, pages 5224–5236 (2016)Cite this article

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Abstract

The transcription factor Twist1 is involved in the epithelial–mesenchymal transition and contributes to cancer metastasis through mostly unknown mechanisms. In colorectal cancer, Twist1 expression is mainly restricted to the tumor stroma. We found that human fibroblast cell lines stably transfected with Twist1 acquired characteristics of activated cancer-associated fibroblasts (CAFs), such as hyperproliferation, an increased ability to migrate and an alignment of the actin cytoskeleton. Further, Twist1-activated fibroblasts promoted increased matrix stiffness. Using quantitative proteomics, we identified palladin and collagen α1(VI) as two major mediators of the Twist1 effects in fibroblast cell lines. Co-immunoprecipitation studies indicated that palladin and Twist1 interact within the nucleus, suggesting that palladin could act as a transcription regulator. Palladin was found to be more relevant for the cellular biomechanical properties, orientation and polarity, and collagen α1(VI) for the migration and invasion capacity, of Twist1-activated fibroblasts. Both palladin and collagen α1(VI) were observed to be overexpressed in colorectal CAFs and to be associated with poor colorectal cancer patient survival and relapse prediction. Our results demonstrate that Twist1-expressing fibroblasts mimic the properties of CAFs present at the tumor invasive front, which likely explains the prometastatic activities of Twist1. Twist1 appears to require both palladin and collagen α1(VI) as downstream effectors for its prometastatic effects, which could be future therapeutic targets in cancer metastasis.

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Acknowledgements

IG-P was supported by a contract S2010/BMD-2344/ Colomics2 from the Comunidad de Madrid. ST was a recipient of a Juan de la Cierva programme. RAB was supported by a grant to established research groups of the Asociación Española Contra el Cáncer (AECC). ML-L was a recipient of a ProteoRed contract. AP-G and BE-P were FPI fellows from the Ministry of Economy and Competitiveness (MINECO). This research was supported by grants to established research groups of the S2010/BMD-2344/Colomics2 from the Comunidad de Madrid, ‘Asociación Española Contra el Cancer (AECC)’, BIO2012-31023 from the MINECO, PRB2 (IPT13/0001-ISCIII-SGEFI/FEDER), RD12/0036/0041 and RD12/0036/0021 from the Instituto de Salud Carlos III-FEDER.

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Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain

    I García-Palmero, S Torres, R A Bartolomé, A Peláez-García, B Escudero-Paniagua & J I Casal

  2. Instituto de Investigaciones Biomédicas ‘Alberto Sols’ (CSIC-UAM), IdiPAZ, Madrid, Spain

    M J Larriba & A Muñoz

  3. Proteomics Core Facility, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain

    M Lopez-Lucendo

  4. Hospital Universitario Puerta de Hierro, Majadahonda, Madrid, Spain

    C Peña

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García-Palmero, I., Torres, S., Bartolomé, R. et al. Twist1-induced activation of human fibroblasts promotes matrix stiffness by upregulating palladin and collagen α1(VI). Oncogene 35, 5224–5236 (2016). https://doi.org/10.1038/onc.2016.57

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