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Aryl-hydrocarbon receptor-interacting protein regulates tumorigenic and metastatic properties of colorectal cancer cells driving liver metastasis

Abstract

Background

Liver metastasis is the primary cause of colorectal cancer (CRC)-associated death. Aryl-hydrocarbon receptor-interacting protein (AIP), a putative positive intermediary in aryl-hydrocarbon receptor-mediated signalling, is overexpressed in highly metastatic human KM12SM CRC cells and other highly metastatic CRC cells.

Methods

Meta-analysis and immunohistochemistry were used to assess the relevance of AIP. Cellular functions and signalling mechanisms mediated by AIP were assessed by gain-of-function experiments and in vitro and in vivo experiments.

Results

A significant association of high AIP expression with poor CRC patients’ survival was observed. Gain-of-function and quantitative proteomics experiments demonstrated that AIP increased tumorigenic and metastatic properties of isogenic KM12C (poorly metastatic) and KM12SM (highly metastatic to the liver) CRC cells. AIP overexpression dysregulated epithelial-to-mesenchymal (EMT) markers and induced several transcription factors and Cadherin-17 activation. The former induced the signalling activation of AKT, SRC and JNK kinases to increase adhesion, migration and invasion of CRC cells. In vivo, AIP expressing KM12 cells induced tumour growth and liver metastasis. Furthermore, KM12C (poorly metastatic) cells ectopically expressing AIP became metastatic to the liver.

Conclusions

Our data reveal new roles for AIP in regulating proteins associated with cancer and metastasis to induce tumorigenic and metastatic properties in colon cancer cells driving liver metastasis.

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Fig. 1: AIP overexpression is associated with colorectal cancer KM12SM liver metastatic cells and to poor survival of colorectal cancer patients.
Fig. 2: AIP-ectopic expression in colorectal cancer cells increases tumorigenic and metastatic properties of colorectal cancer cells.
Fig. 3: Alterations in EMT inducers after AIP-ectopic expression in colorectal cancer cells.
Fig. 4: Mass spectrometry analysis of protein alterations modulated by AIP overexpression in CRC cells.
Fig. 5: AIP-ectopic expression induces tumour growth and liver metastasis in KM12 cells.

Data availability

All data generated or analysed during this study are included in this published article and its supplementary information files. The Mass Spectrometry data were deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD031119. Other data are available from the corresponding author on reasonable request.

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Funding

This work was supported by the financial support of the PI17CIII/00045 and PI20CIII/00019 grants from the AES-ISCIII program to RB. J Hendrix acknowledges funding by UH-BOF (BOF20TT06). J Hofkens acknowledges financial support from the Research Foundation-Flanders (FWO, Grant No. ZW15_09-G0H6316N), the Flemish government through long-term structural funding Methusalem (CASAS2, Meth/15/04) and the MPI as MPI fellow. S.R. acknowledges the financial support of the KU Leuven through the internal C1 funding (KU Leuven (C14/16/053)). GSF is the recipient of a predoctoral contract (grant number 1193818 N) supported by The Flanders Research Foundation (FWO). The FPU predoctoral contract to AMC is supported by the Spanish Ministerio de Educación, Cultura y Deporte.

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Authors

Contributions

GSF: data curation; formal analysis, investigation, writing—original draft and writing—review and editing. AMC: data curation, investigation, writing—original draft and writing—review and editing. MSM: data curation and investigation. APG: formal analysis, investigation, methodology, supervision and writing—review and editing. MJFA: formal analysis, methodology, supervision and writing—review and editing. PP: investigation and writing—review. MAN: investigation and writing—review. MM: methodology, supervision and writing—review. J Hendrix: supervision and writing—review. DH: methodology, supervision and writing—review. RAB: conceptualisation, investigation, supervision and writing—review & editing. J Hofkens: conceptualisation, investigation, supervision, writing—review & editing and funding. SR: conceptualisation, formal analysis, investigation, writing—original draft, writing—review and editing and funding. RB: conceptualisation, formal analysis, investigation, writing—original draft, writing—review and editing and funding.

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Correspondence to Susana Rocha or Rodrigo Barderas.

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All animal experiments were performed under the guidelines of the Instituto de Salud Carlos III Institutional Animal Care Committee. All experimental protocols were approved by the Committee (Proex 285/19). This article does not contain any studies with human participants by any of the authors out of public databases. However, immunohistochemical analyses of human tumour specimens were in compliance with ethical standards and approved by the Hospital Clínico San Carlos Ethics Board; and informed consent was obtained from all individual participants included in the study.

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Solís-Fernández, G., Montero-Calle, A., Sánchez-Martínez, M. et al. Aryl-hydrocarbon receptor-interacting protein regulates tumorigenic and metastatic properties of colorectal cancer cells driving liver metastasis. Br J Cancer 126, 1604–1615 (2022). https://doi.org/10.1038/s41416-022-01762-1

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