Original ArticlesLaminin 521 enhances self-renewal via STAT3 activation and promotes tumor progression in colorectal cancer
Introduction
Although clinical outcomes have steadily improved over the last few decades for patients with colorectal cancer, survival remains poor for those who have developed metastatic disease to distant sites such as the liver, lung or peritoneum [1]. Characterizing cellular and molecular processes that drive metastasis progression therefore represents an essential step to better prevent or target metastatic disease. Previous studies have demonstrated that the development of metastases requires tumor cells to display invasive properties, enabling their dissemination from primary sites and transit through the lymphatic and/or blood stream, as well as survival and self-renewing properties that will drive their ability to initiate metastasis growth and colonize the distant site [2,3]. Invasion and self-renewal are both sensitive to microenvironmental cues provided by cellular and non-cellular actors of the microenvironment. Among the latter, extracellular matrix molecules have been described as instrumental modulators of cell motility, invasion, and stemness [4]. In particular, remodeling of basement membrane proteins such as laminins during cancer development is thought to alter interaction of these proteins with receptors located at the surface of cancer cells, thereby leading to phenotypic modifications [5].
Laminins are large multimeric basement membrane proteins, with functional units constituted through the assembly of one alpha, one beta and one gamma isoform [6]. In the present work we analyzed the contribution of laminin alpha 5 (LAMA5), and more specifically of the laminin-521 isoform (LN521, constituted by one alpha 5, one beta-2 and one gamma-1 chain), in the regulation of self-renewal and invasion of colorectal cancer cells. Laminin-521 (LN521) was previously shown to be important in the maintenance of embryonic and induced-pluripotent stem cells [7,8], and we hypothesized that exposure to LN521 could also promote self-renewal in colorectal cancer cells. Since long-term self-renewal is a hallmark of cancer stem cells (CSCs) and contributes to their driving role in cancer progression and poor clinical outcomes [9,10], we also investigated whether LN521 may play a role in regulating phenotypic and clinical features that are classically associated with metastatic development, such as cell migration, invasion and patient survival.
Section snippets
Materials and methods
Immunohistochemical staining, Extreme Limiting Dilution Analysis (ELDA), Protein and RNA quantitation, in vivo liver metastasis induction in immunocompromised mice, as well as analysis of associations between laminin expression and CRC patient survival, were performed as previously described [11,12]. Details for these procedures are provided under Supplemental Materials and Methods.
Laminin-521 promotes self-renewal and invasion of CRC cells
To determine whether exposure to LN521 impacted on the self-renewal of colorectal cancer cells, we first exposed human CRC cells to various laminin isoforms and subsequently resuspended and seeded them into an extreme limiting dilution analysis (ELDA) in suspension. The ability of these cells to generate at least one colonosphere per well under all limiting dilution conditions was quantified and the frequency of self-renewing cells was calculated as described previously [11,13] using the //bioinf.wehi.edu.au/software/elda/
Discussion
In the present study we characterized the promoting role of laminin-521 on colorectal cancer cell self-renewal and invasion and found that endogenous production of the laminin alpha 5 chain contributed to this effect. We demonstrated that integrins α3β1 and α6β1 are likely mediators of the self-renewal promotion induced by laminin alpha 5 and that downstream activation of STAT3 signaling is required for the promotion of self-renewal by LN521. Finally, we also found that colorectal cancer liver
Funding
This work was supported by the National Health and Medical Research Council of Australia [grants number GNT1049561 and GNT1069024].
CRediT authorship contribution statement
Yan Qin: Conceptualization, Methodology, Investigation, Formal analysis, Visualization, Writing - original draft. Carolyn Shembrey: Investigation, Validation, Writing - review & editing. Jai Smith: Investigation, Validation, Visualization. Sophie Paquet-Fifield: Investigation, Methodology. Corina Behrenbruch: Methodology, Resources. Laura M. Beyit: Investigation. Benjamin N.J. Thomson: Resources. Alexander G. Heriot: Resources, Conceptualization. Yuan Cao: Supervision, Formal analysis, Writing
Declaration of competing interest
The authors have no conflict of interest to declare in relation with this work.
References (51)
- et al.
Colorectal cancer
Lancet
(2014) - et al.
Tumor metastasis: molecular insights and evolving paradigms
Cell
(2011) Laminins and cancer stem cells: partners in crime?
Semin. Canc. Biol.
(2017)alpha-5 laminin synthesized by human pluripotent stem cells promotes self-renewal
Stem Cell Rep.
(2015)- et al.
ELDA: extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays
J. Immunol. Methods
(2009) Laminin-332 sustains chemoresistance and quiescence as part of the human hepatic cancer stem cell niche
J. Hepatol.
(2016)Long-term self-renewal of human ES/iPS-derived hepatoblast-like cells on human laminin 111-coated dishes
Stem Cell Rep.
(2013)Ligand-binding specificities of laminin-binding integrins: a comprehensive survey of laminin-integrin interactions using recombinant alpha3beta1, alpha6beta1, alpha7beta1 and alpha6beta4 integrins
Matrix Biol.
(2006)Epithelial gp130/Stat3 functions: an intestinal signaling node in health and disease
Semin. Immunol.
(2014)Maintenance of clonogenic KIT(+) human colon tumor cells requires secretion of stem cell factor by differentiated tumor cells
Gastroenterology
(2015)
CD44v6 is a marker of constitutive and reprogrammed cancer stem cells driving colon cancer metastasis
Cell Stem Cell
Collaboration of 3D context and extracellular matrix in the development of glioma stemness in a 3D model
Biomaterials
Melanoma cells produce multiple laminin isoforms and strongly migrate on alpha5 laminin(s) via several integrin receptors
Exp. Cell Res.
Immuno-proteomic discovery of tumor tissue autoantigens identifies olfactomedin 4, CD11b, and integrin alpha-2 as markers of colorectal cancer with liver metastases
J Proteomics
Association of CD98, integrin beta1, integrin beta3 and Fak with the progression and liver metastases of colorectal cancer
Pathol. Res. Pract.
The JAK/STAT3 axis: a comprehensive drug target for solid malignancies
Semin. Canc. Biol.
Laminin alpha 5, a major transcript of normal and malignant rat liver epithelial cells, is differentially expressed in developing and adult liver
Exp. Cell Res.
alpha1- and alpha5-containing laminins regulate the development of bile ducts via beta1 integrin signals
J. Biol. Chem.
To differentiate or not--routes towards metastasis
Nat. Rev. Canc.
Remodelling the extracellular matrix in development and disease
Nat. Rev. Mol. Cell Biol.
The laminin family
Cell Adhes. Migrat.
Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment
Nat. Commun.
Self-renewal as a therapeutic target in human colorectal cancer
Nat. Med.
Targeting the cancer initiating cell: the ultimate target for cancer therapy
Curr. Pharmaceut. Des.
Autocrine secretion of progastrin promotes the survival and self-renewal of colon cancer stem-like cells
Cancer Res.
Cited by (23)
Pan-caner analysis identifies PSMA7 as a targets for amplification at 20q13.33 in tumorigenesis
2024, Scientific Reports