Skip to main content

Advertisement

SpringerLink
Log in

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

  • Article
  • Published:
British Journal of Cancer Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

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.

Similar content being viewed by others

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.

References

  1. Chaffer CL, Weinberg RA. A perspective on cancer cell metastasis. Science. 2011;331:1559–64.

    Article  CAS  PubMed  Google Scholar 

  2. Torres S, Bartolome RA, Mendes M, Barderas R, Fernandez-Acenero MJ, Pelaez-Garcia A, et al. Proteome profiling of cancer-associated fibroblasts identifies novel proinflammatory signatures and prognostic markers for colorectal cancer. Clin Cancer Res. 2013;19:6006–19.

    Article  CAS  PubMed  Google Scholar 

  3. Quail DF, Joyce JA. Microenvironmental regulation of tumor progression and metastasis. Nat Med. 2013;19:1423–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Cai Z, Chiu JF, He QY. Application of proteomics in the study of tumor metastasis. Genomics Proteom Bioinforma. 2004;2:152–66.

    Article  CAS  Google Scholar 

  5. Aleckovic M, Wei Y, LeRoy G, Sidoli S, Liu DD, Garcia BA, et al. Identification of Nidogen 1 as a lung metastasis protein through secretome analysis. Genes Dev. 2017;31:1439–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Song P, Bao H, Yu Y, Xue Y, Yun D, Zhang Y, et al. Comprehensive profiling of metastasis-related proteins in paired hepatocellular carcinoma cells with different metastasis potentials. Proteom Clin Appl. 2009;3:841–52.

    Article  CAS  Google Scholar 

  7. Sun B, Zhang S, Zhang D, Li Y, Zhao X, Luo Y, et al. Identification of metastasis-related proteins and their clinical relevance to triple-negative human breast cancer. Clin Cancer Res. 2008;14:7050–9.

    Article  CAS  PubMed  Google Scholar 

  8. Barderas R, Mendes M, Torres S, Bartolome RA, Lopez-Lucendo M, Villar-Vazquez R, et al. In-depth characterization of the secretome of colorectal cancer metastatic cells identifies key proteins in cell adhesion, migration, and invasion. Mol Cell Proteom. 2013;12:1602–20.

    Article  CAS  Google Scholar 

  9. Mendes M, Pelaez-Garcia A, Lopez-Lucendo M, Bartolome RA, Calvino E, Barderas R, et al. Mapping the spatial proteome of metastatic cells in colorectal cancer. Proteomics. 2017;17:1700094.

  10. Morikawa K, Walker SM, Jessup JM, Fidler IJ. In vivo selection of highly metastatic cells from surgical specimens of different primary human colon carcinomas implanted into nude mice. Cancer Res. 1988;48:1943–8.

    CAS  PubMed  Google Scholar 

  11. Morikawa K, Walker SM, Nakajima M, Pathak S, Jessup JM, Fidler IJ. Influence of organ environment on the growth, selection, and metastasis of human colon carcinoma cells in nude mice. Cancer Res. 1988;48:6863–71.

    CAS  PubMed  Google Scholar 

  12. Li A, Varney ML, Singh RK. Constitutive expression of growth regulated oncogene (gro) in human colon carcinoma cells with different metastatic potential and its role in regulating their metastatic phenotype. Clin Exp Metastasis. 2004;21:571–9.

    Article  CAS  PubMed  Google Scholar 

  13. Kuniyasu H, Ohmori H, Sasaki T, Sasahira T, Yoshida K, Kitadai Y, et al. Production of interleukin 15 by human colon cancer cells is associated with induction of mucosal hyperplasia, angiogenesis, and metastasis. Clin Cancer Res. 2003;9:4802–10.

    CAS  PubMed  Google Scholar 

  14. Calon A, Espinet E, Palomo-Ponce S, Tauriello DV, Iglesias M, Cespedes MV, et al. Dependency of colorectal cancer on a TGF-beta-driven program in stromal cells for metastasis initiation. Cancer Cell. 2012;22:571–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Barderas R, Bartolome RA, Fernandez-Acenero MJ, Torres S, Casal JI. High expression of IL-13 receptor alpha2 in colorectal cancer is associated with invasion, liver metastasis, and poor prognosis. Cancer Res. 2012;72:2780–90.

    Article  CAS  PubMed  Google Scholar 

  16. Hegde P, Qi R, Gaspard R, Abernathy K, Dharap S, Earle-Hughes J, et al. Identification of tumor markers in models of human colorectal cancer using a 19,200-element complementary DNA microarray. Cancer Res. 2001;61:7792–7.

    CAS  PubMed  Google Scholar 

  17. Ellens KW, Christian N, Singh C, Satagopam VP, May P, Linster CL. Confronting the catalytic dark matter encoded by sequenced genomes. Nucleic Acids Res. 2017;45:11495–514.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Perdigao N, Heinrich J, Stolte C, Sabir KS, Buckley MJ, Tabor B, et al. Unexpected features of the dark proteome. Proc Natl Acad Sci USA. 2015;112:15898–903.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Tang Z, Kang B, Li C, Chen T, Zhang Z. GEPIA2: an enhanced web server for large-scale expression profiling and interactive analysis. Nucleic Acids Res. 2019;47:W556–W560.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Garranzo-Asensio M, San Segundo-Acosta P, Poves C, Fernandez-Acenero MJ, Martinez-Useros J, Montero-Calle A, et al. Identification of tumor-associated antigens with diagnostic ability of colorectal cancer by in-depth immunomic and seroproteomic analysis. J Proteom. 2020;214:103635.

    Article  CAS  Google Scholar 

  21. Garranzo-Asensio M, San Segundo-Acosta P, Martinez-Useros J, Montero-Calle A, Fernandez-Acenero MJ, Haggmark-Manberg A, et al. Identification of prefrontal cortex protein alterations in Alzheimer’s disease. Oncotarget. 2018;9:10847–67.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Barderas R, Villar-Vazquez R, Fernandez-Acenero MJ, Babel I, Pelaez-Garcia A, Torres S, et al. Sporadic colon cancer murine models demonstrate the value of autoantibody detection for preclinical cancer diagnosis. Sci Rep. 2013;3:2938.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Bartolome RA, Barderas R, Torres S, Fernandez-Acenero MJ, Mendes M, Garcia-Foncillas J, et al. Cadherin-17 interacts with alpha2beta1 integrin to regulate cell proliferation and adhesion in colorectal cancer cells causing liver metastasis. Oncogene. 2014;33:1658–69.

    Article  CAS  PubMed  Google Scholar 

  24. Pelaez-Garcia A, Barderas R, Torres S, Hernandez-Varas P, Teixido J, Bonilla F, et al. FGFR4 role in epithelial-mesenchymal transition and its therapeutic value in colorectal cancer. PLoS ONE. 2013;8:e63695.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Barderas R, Shochat S, Timmerman P, Hollestelle MJ, Martinez-Torrecuadrada JL, Hoppener JW, et al. Designing antibodies for the inhibition of gastrin activity in tumoral cell lines. Int J Cancer. 2008;122:2351–9.

    Article  CAS  PubMed  Google Scholar 

  26. Borowicz S, Van Scoyk M, Avasarala S, Karuppusamy Rathinam MK, Tauler J, Bikkavilli RK, et al. The soft agar colony formation assay. J Vis Exp. 2014;92:e51998.

  27. Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9:671–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Exploring data normalization and analysis in large TMT experimental designs [https://pwilmart.github.io/IRS_normalization/understanding_IRS.html].

  29. Kubens BS, Zanker KS. Differences in the migration capacity of primary human colon carcinoma cells (SW480) and their lymph node metastatic derivatives (SW620). Cancer Lett. 1998;131:55–64.

    Article  CAS  PubMed  Google Scholar 

  30. Araki K, Shimura T, Suzuki H, Tsutsumi S, Wada W, Yajima T, et al. E/N-cadherin switch mediates cancer progression via TGF-beta-induced epithelial-to-mesenchymal transition in extrahepatic cholangiocarcinoma. Br J Cancer. 2011;105:1885–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, et al. STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 2019;47:D607–D613.

    Article  CAS  PubMed  Google Scholar 

  32. Griss J, Viteri G, Sidiropoulos K, Nguyen V, Fabregat A, Hermjakob H. ReactomeGSA—efficient Multi-Omics comparative pathway analysis. Mol Cell Proteom. 2020;19:2115–25.

    Article  CAS  Google Scholar 

  33. Tian X, Yang C, Yang L, Sun Q, Liu N. PTPRF as a novel tumor suppressor through deactivation of ERK1/2 signaling in gastric adenocarcinoma. Onco Targets Ther. 2018;11:7795–803.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Fang WK, Liao LD, Li LY, Xie YM, Xu XE, Zhao WJ, et al. Down-regulated desmocollin-2 promotes cell aggressiveness through redistributing adherens junctions and activating beta-catenin signalling in oesophageal squamous cell carcinoma. J Pathol. 2013;231:257–70.

    Article  CAS  PubMed  Google Scholar 

  35. Bujko M, Kober P, Mikula M, Ligaj M, Ostrowski J, Siedlecki JA. Expression changes of cell-cell adhesion-related genes in colorectal tumors. Oncol Lett. 2015;9:2463–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Bajpai R, Nagaraju GP. Specificity protein 1: its role in colorectal cancer progression and metastasis. Crit Rev Oncol Hematol. 2017;113:1–7.

    Article  PubMed  Google Scholar 

  37. Wang H, Li K, Mei Y, Huang X, Li Z, Yang Q, et al. Sp1 suppresses miR-3178 to promote the metastasis invasion cascade via upregulation of TRIOBP. Mol Ther Nucleic Acids. 2018;12:1–11.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  38. Khodarev NN, Roach P, Pitroda SP, Golden DW, Bhayani M, Shao MY, et al. STAT1 pathway mediates amplification of metastatic potential and resistance to therapy. PLoS ONE. 2009;4:e5821.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  39. Basak O, van de Born M, Korving J, Beumer J, van der Elst S, van Es JH, et al. Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele. EMBO J 2014;33:2057–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Coopmans EC, Muhammad A, Daly AF, de Herder WW, van Kemenade FJ, Beckers A, et al. The role of AIP variants in pituitary adenomas and concomitant thyroid carcinomas in the Netherlands: a nationwide pathology registry (PALGA) study. Endocrine. 2020;68:640–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Georgitsi M, Karhu A, Winqvist R, Visakorpi T, Waltering K, Vahteristo P, et al. Mutation analysis of aryl hydrocarbon receptor interacting protein (AIP) gene in colorectal, breast, and prostate cancers. Br J Cancer. 2007;96:352–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Diaz Del Arco C, Estrada Munoz L, Barderas Manchado R, Pelaez Garcia A, Ortega Medina L, Molina Roldan E, et al. Prognostic role of Aryl hydrocarbon receptor interacting protein (AIP) immunohistochemical expression in patients with resected gastric carcinomas. Pathol Oncol Res. 2020;26:2641–50.

    Article  CAS  PubMed  Google Scholar 

  43. Fernández-Aceñero MJ, Barderas R, Peláez-García A, Martínez-Useros J, Díez-Valladares L, Ortega-Medina L, et al. Aryl hydrocarbon receptor interacting protein (AIP) significantly influences prognosis of pancreatic carcinoma. Annal Diagnostic Pathol. 2021;53:151742.

  44. Barry S, Carlsen E, Marques P, Stiles CE, Gadaleta E, Berney DM, et al. Tumor microenvironment defines the invasive phenotype of AIP-mutation-positive pituitary tumors. Oncogene. 2019;38:5381–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Vasilev V, Daly AF, Trivellin G, Stratakis CA, Zacharieva S, Beckers A. Hereditary endocrine tumours: current state-of-the-art and research opportunities: the roles of AIP and GPR101 in familial isolated pituitary adenomas (FIPA). Endocr Relat Cancer. 2020;27:T77–T86.

    Article  CAS  PubMed  Google Scholar 

  46. Leontiou CA, Gueorguiev M, van der Spuy J, Quinton R, Lolli F, Hassan S, et al. The role of the aryl hydrocarbon receptor-interacting protein gene in familial and sporadic pituitary adenomas. J Clin Endocrinol Metab. 2008;93:2390–401.

    Article  CAS  PubMed  Google Scholar 

  47. Trivellin G, Korbonits M. AIP and its interacting partners. J Endocrinol. 2011;210:137–55.

    Article  CAS  PubMed  Google Scholar 

  48. Rothhammer V, Quintana FJ. The aryl hydrocarbon receptor: an environmental sensor integrating immune responses in health and disease. Nat Rev Immunol. 2019;19:184–97.

    Article  CAS  PubMed  Google Scholar 

  49. Meyer BK, Pray-Grant MG, Vanden Heuvel JP, Perdew GH. Hepatitis B virus X-associated protein 2 is a subunit of the unliganded aryl hydrocarbon receptor core complex and exhibits transcriptional enhancer activity. Mol Cell Biol. 1998;18:978–88.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer. Mol Oncol. 2018;12:3–20.

    Article  PubMed  Google Scholar 

  51. Markman B, Javier Ramos F, Capdevila J, Tabernero J. EGFR and KRAS in colorectal cancer. Adv Clin Chem. 2010;51:71–119.

    Article  CAS  PubMed  Google Scholar 

  52. Sigismund S, Argenzio E, Tosoni D, Cavallaro E, Polo S, Di Fiore PP. Clathrin-mediated internalization is essential for sustained EGFR signaling but dispensable for degradation. Dev Cell. 2008;15:209–19.

    Article  CAS  PubMed  Google Scholar 

Download references

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.

Author information

Author notes

  1. These authors contributed equally: Ana Montero-Calle, Maricruz Sánchez-Martínez.

  2. These authors jointly supervised this work: Rubén A. Bartolomé, Rodrigo Barderas.

Authors and Affiliations

  1. Chronic Disease Programme (UFIEC), Instituto de Salud Carlos III, Majadahonda, E-28220, Madrid, Spain

    Guillermo Solís-Fernández, Ana Montero-Calle, Maricruz Sánchez-Martínez, Miren Alonso-Navarro & Rodrigo Barderas

  2. Molecular Imaging and Photonics Division, Chemistry Department, Faculty of Sciences, KU Leuven, Celestijnenlaan 200F, 3001 Heverlee, Leuven, Belgium

    Guillermo Solís-Fernández, Johan Hofkens & Susana Rocha

  3. Molecular Pathology and Therapeutic Targets Group, La Paz University Hospital (IdiPAZ), E-28046, Madrid, Spain

    Alberto Peláez-García, Marta Mendiola & David Hardisson

  4. Surgical Pathology Department, Hospital Universitario Clínico San Carlos, E-28040, Madrid, Spain

    María Jesús Fernández-Aceñero

  5. Unidades Centrales, Instituto de Salud Carlos III, Majadahonda, E-28220, Madrid, Spain

    Pilar Pallarés

  6. Dynamic Bioimaging Lab, Advanced Optical Microscopy Centre and Biomedical Research Institute, Hasselt University, Agoralaan C (BIOMED), 3590 Diepenbeek, Hasselt, Belgium

    Jelle Hendrix

  7. Centro de Investigaciones Biológicas, CSIC, E-28040, Madrid, Spain

    Rubén A. Bartolomé

  8. Max Planck Institute for Polymer Research, Ackermannweg 10, 55128, Mainz, Germany

    Johan Hofkens

Authors
  1. Guillermo Solís-Fernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ana Montero-Calle
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maricruz Sánchez-Martínez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alberto Peláez-García
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. María Jesús Fernández-Aceñero
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pilar Pallarés
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Miren Alonso-Navarro
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Marta Mendiola
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jelle Hendrix
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. David Hardisson
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Rubén A. Bartolomé
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Johan Hofkens
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Susana Rocha
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Rodrigo Barderas
    View author publications

    You can also search for this author in PubMed Google Scholar

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.

Corresponding authors

Correspondence to Susana Rocha or Rodrigo Barderas.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

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.

Consent to publish

All authors of this article gave consent for the publication of the manuscript.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41416-022-01762-1

  • Springer Nature Limited

This article is cited by

Search

Navigation