Direct cell–cell interaction enhances pro-MMP-2 production and activation in co-culture of laryngeal cancer cells and fibroblasts: involvement of EMMPRIN and MT1-MMP
Introduction
Degradation of extracellular matrix (ECM) components by matrix metalloproteinases (MMPs) is critical for tumor cell invasion and metastasis [1], [2]. Since disruption of the basement membrane is a crucial step for tumor progression, of MMPs, matrix metalloproteinases-2 (MMP-2) has been noted because of its activity to degrade type IV collagen, a major component of the basement membrane, and membrane type 1-matrix metalloproteinase (MT1-MMP) has been recognized as a key factor for pro-MMP-2 activation [3]. Recent studies have revealed that tumor cells utilize MMPs such as MMP-2 produced by neighboring stromal cells including fibroblasts rather than by tumor cells themselves [4], [5] for tumor progression, invasion, and metastasis, and that tumor cells can stimulate MMP production by stromal cells via soluble factors such as cytokines [6], [7], [8], [9] or through cell–cell interaction mediated by cell adhesion molecules such as EMMPRIN [10].
In addition to several humoral factors such as cytokines affecting the production and activation of ECM-degrading proteases including MMPs in tumor cells and/or stromal cells, EMMPRIN/CD147 has been noted to be highly expressed on tumor cell surface and involved in tumor progression by inducing the production of MMPs including MMP-2 by neighboring stromal cells [11], [12], [13]. EMMPRIN belongs to immunoglobulin superfamily, is referred to also as TCSF [14], basigin [15], neurotherin [16], or M6 antigen [17], and is a transmembrane glycoprotein with two extracellular domains and a short cytoplasmic domain [14], [18]. It has been reported that EMMPRIN is enriched in variety of human carcinomas [19], [20], [21] and contributes to tumor invasion and metastasis by stimulating fibroblasts nearby to secrete an increased amount of MMPs [4], [22]. However, very few attempts have been made to clarify an involvement of EMMPRIN in progression of head and neck cancers.
To explore the mechanism of tumor progression, in this study, we examined an expression of MMPs and EMMPRIN in cultured HEp-2 cells derived from a human laryngeal epidermoid carcinoma, fibroblasts, and their co-culture, and confirmed the importance of direct cell–cell association between tumor cells and fibroblasts and the contribution of EMMPRIN to stimulation of MMP-2 production and activation from fibroblasts, namely, stromal cells.
Section snippets
Cells and cell culture
HEp-2 cells, a human laryngeal carcinoma cell line, were provided by Cell Resource Center for Biomedical Research Institute of Development, Aging and Cancer, Tohoku University (Sendai, Japan). Human fibroblasts were isolated from the dermis of a healthy subject. Cells were grown in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) in a humidified atmosphere containing 5% CO2 at 37°C. The protocol used in this study for specimen preparation was previously approved
Stimulation of pro-MMP-2 production and activation by co-culture of HEp-2 cells and fibroblasts
It has been reported that the interaction of cancer cells with stromal cells is a pivotal step to produce pro-MMP-2 and/or activate pro-MMP-2 for tumor progression. To explore an involvement of cell–cell interaction in pro-MMP-2 production and/or pro-MMP-2 activation, a cell line HEp-2 derived from a human larynx epidermoid carcinoma was co-cultured with human skin fibroblasts to analyze pro-MMP-2 production and activation. Zymographic analysis of the conditioned media from fibroblast and HEp-2
Discussion
MMPs, a family of zinc-dependent proteinases, are capable to degrade a variety of ECM components, and have various physiological functions in normal tissues [25], [26], whereas the degradation of ECM components by MMPs is a determining step in tumor cell invasion and metastasis [1], [2], [27], [28]. Of MMPs, MMP-2 (gelatinase A) is able to degrade type IV collagen in the basement membrane for tumor progression [1], and therefore pro-MMP-2 expression and activation are important events in many
Acknowledgements
The authors thank Drs. Katsuyuki Imai, Takeya Sato, Da-Ren Wang, and Nobuyo Higashi (Department of Cell Biology and Histology, Akita University School of Medicine) for their valuable discussions. The expert technical support provided by Naosuke Kojima (Department of Cell Biology and Histology, Akita University School of Medicine) was also gratefully appreciated.
References (41)
- et al.
Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation
Cell
(1991) - et al.
Transcriptional and post-transcriptional regulation of 72-kDa gelatinase/type IV collagenase by transforming growth factor-beta 1 in human fibroblasts. Comparisons with collagenase and tissue inhibitor of matrix metalloproteinase gene expression
J. Biol. Chem.
(1991) - et al.
Glioma cell extracellular matrix metalloproteinase inducer (EMMPRIN) (CD147) stimulates production of membrane-type matrix metalloproteinases and activated gelatinase A in co-cultures with brain-derived fibroblasts
Cancer Lett.
(2000) - et al.
Stimulation of matrix metalloproteinase production by recombinant extracellular matrix metalloproteinase inducer from transfected Chinese hamster ovary cells
J. Biol. Chem.
(1997) - et al.
Characterization of the gene for human EMMPRIN, a tumor cell surface inducer of matrix metalloproteinases
Gene
(1998) - et al.
Secretion of metalloproteinases by stimulated capillary endothelial cells. II. Expression of collagenase and stromelysin activities is regulated by endogenous inhibitors
J. Biol. Chem.
(1986) - et al.
Mechanism of cell surface activation of 72-kDa type IV collagenase. Isolation of the activated form of the membrane metalloprotease
J. Biol. Chem.
(1995) - et al.
Tumor cell interactions with the extracellular matrix during invasion and metastasis
Annu. Rev. Cell Biol.
(1993) - et al.
A matrix metalloproteinase expressed on the surface of invasive tumour cells
Nature
(1994) - et al.
Basigin (CD147) is expressed on melanoma cells and induces tumor cell invasion by stimulating production of matrix metalloproteinases by fibroblasts
Int. J. Cancer
(2002)
Stromal expression of 72 kDa type IV collagenase (MMP-2) and TIMP-2 mRNAs in colorectal neoplasia
Am. J. Pathol.
Regulation of the Mr 72,000 type IV collagenase by the type I insulin-like growth factor receptor
Cancer Res.
Independent expression and cellular processing of Mr 72,000 type IV collagenase and interstitial collagenase in human tumorigenic cell lines
Cancer Res.
Transforming growth factor-beta 1 stimulates glomerular mesangial cell synthesis of the 72-kD type IV collagenase
Am. J. Pathol.
Coordinate increase in collagenase mRNA and enzyme levels in human fibroblasts treated with the tumor cell factor, TCSF
Biochem. Int.
Basigin (murine EMMPRIN) stimulates matrix metalloproteinase production by fibroblasts
J. Cell. Physiol.
The human tumor cell-derived collagenase stimulatory factor (renamed EMMPRIN) is a member of the immunoglobulin superfamily
Cancer Res.
Basigin, a new member of the immunoglobulin superfamily: genes in different mammalian species, glycosylation changes in the molecule from adult organs and possible variation in the N-terminal sequences
Cell Struct. Funct.
Neurothelin: amino acid sequence, cell surface dynamics and actin colocalization
Eur. J. Cell Biol.
Human leukocyte activation antigen M6, a member of the Ig superfamily, is the species homologue of rat OX-47, mouse basigin, and chicken HT7 molecule
J. Immunol.
Cited by (103)
The Role of Matrix Metalloproteinases in Development, Repair, and Destruction of the Lungs
2017, Progress in Molecular Biology and Translational ScienceCitation Excerpt :In addition, the MMP inducer CD147 is present during lung development,23 though its expression decreases to low levels in normal adult lung.24 Studies have shown that CD147 increases the expression of MMP1, MMP2, MMP3, MMP9, and MMP14 in the lung,25–27 and early increases in CD147 are seen in models of lung damage, such as bleomycin-induced lung injury28 and ventilator-associated acute lung injury (ALI).29 MMP2, MMP14, and CD147 are constitutively expressed in all stages of mouse lung development, though their expression tapers off once lung development is complete.12
Nanoparticle-mediated siRNA delivery assessed in a 3D co-culture model simulating prostate cancer bone metastasis
2016, International Journal of PharmaceuticsCitation Excerpt :It was not possible to characterise enzyme secretion from the individual cell types in the co-culture system. Therefore secretion of total MMP levels into the supernatant was analysed (Suzuki et al., 2004). As previously reported, PC3 cells secreted MMP9 under normal monoculture conditions whereas LNCaP cells did not, correlating with the more invasive phenotype of the PC3 cell line (Fig. 4(a) and (c) vs (b) and (d)) (Fitzgerald et al., 2015a).