Tumour Cells Express Functional Lymphatic Endothelium-Specific Hyaluronan Receptor In Vitro and In Vivo: Lymphatic Mimicry Promotes Oral Oncogenesis?

Lymphatic metastasis represents the main route of tumour cell dissemination in oral squamous cell carcinoma (OSCC). Yet, there are no FDA-approved therapeutics targeting cancer-related lymphangiogenesis to date. The lymphatic vessel endothelial hyaluronic acid receptor 1 (LYVE-1), a specific lymphatic marker, is associated with poor survival in OSCC patients. In this study, we present a potential novel mechanism of lymphatic metastasis in OSCC—lymphatic mimicry (LM), a process whereby tumour cells form cytokeratin+/LYVE-1+, but podoplanin-negative, mosaic endothelial-like vessels. LM was detected in one-third (20/57; 35.08%) of randomly selected OSCC patients. The LM-positive patients had shorter overall survival (OS) compared to LM-negative group albeit not statistically significant. Highly-metastatic tumour cells formed distinct LM structures in vitro and in vivo. Importantly, the siRNA-mediated knockdown of LYVE-1 not only impaired tumour cell migration but also blunted their capacity to form LM-vessels in vitro and reduced tumour metastasis in vivo. Together, our findings uncovered, to our knowledge, a previously unknown expression and function of LYVE-1 in OSCC, whereby tumour cells could induce LM formation and promote lymphatic metastasis. Finally, more detailed studies on LM are warranted to better define this phenomenon in the future. These studies could benefit the development of targeted therapeutics for blocking tumour-related lymphangiogenesis.Peer reviewe

The interplay of matrix metalloproteinase-8, transforming growth factor-beta 1 and vascular endothelial growth factor-C cooperatively contributes to the aggressiveness of oral tongue squamous cell carcinoma

Background: Matrix metalloproteinase-8 (MMP-8) has oncosuppressive properties in various cancers. We attempted to assess MMP-8 function in oral tongue squamous cell carcinoma (OTSCC). Methods: MMP-8 overexpressing OTSCC cells were used to study the effect of MMP-8 on proliferation, apoptosis, migration, invasion and gene and protein expression. Moreover, MMP-8 functions were assessed in the orthotopic mouse tongue cancer model and by immunohistochemistry in patient samples. Results: MMP-8 reduced the invasion and migration of OTSCC cells and decreased the expression of MMP-1, cathepsin-K and vascular endothelial growth factor-C (VEGF-C). VEGF-C was induced by transforming growth factor-beta 1 (TGF-beta 1) in control cells, but not in MMP-8 overexpressing cells. In human OTSCC samples, low MMP-8 in combination with high VEGF-C was an independent predictor of poor cancer-specific survival. TGF-beta 1 treatment also restored the migration of MMP-8 overexpressing cells to the level of control cells. In mouse tongue cancer, MMP-8 did not inhibit metastasis, possibly because it was eliminated in the peripheral carcinoma cells. Conclusions: The suppressive effects of MMP-8 in OTSCC may be mediated through interference of TGF-beta 1 and VEGF-C function and altered proteinase expression. Together, low MMP-8 and high VEGF-C expression have strong independent prognostic value in OTSCC.Peer reviewe

The interplay of matrix metalloproteinase-8, transforming growth factor-beta 1 and vascular endothelial growth factor-C cooperatively contributes to the aggressiveness of oral tongue squamous cell carcinoma

Background: Matrix metalloproteinase-8 (MMP-8) has oncosuppressive properties in various cancers. We attempted to assess MMP-8 function in oral tongue squamous cell carcinoma (OTSCC). Methods: MMP-8 overexpressing OTSCC cells were used to study the effect of MMP-8 on proliferation, apoptosis, migration, invasion and gene and protein expression. Moreover, MMP-8 functions were assessed in the orthotopic mouse tongue cancer model and by immunohistochemistry in patient samples. Results: MMP-8 reduced the invasion and migration of OTSCC cells and decreased the expression of MMP-1, cathepsin-K and vascular endothelial growth factor-C (VEGF-C). VEGF-C was induced by transforming growth factor-beta 1 (TGF-beta 1) in control cells, but not in MMP-8 overexpressing cells. In human OTSCC samples, low MMP-8 in combination with high VEGF-C was an independent predictor of poor cancer-specific survival. TGF-beta 1 treatment also restored the migration of MMP-8 overexpressing cells to the level of control cells. In mouse tongue cancer, MMP-8 did not inhibit metastasis, possibly because it was eliminated in the peripheral carcinoma cells. Conclusions: The suppressive effects of MMP-8 in OTSCC may be mediated through interference of TGF-beta 1 and VEGF-C function and altered proteinase expression. Together, low MMP-8 and high VEGF-C expression have strong independent prognostic value in OTSCC.Peer reviewe

Matrix metalloproteinase 8 as a marker and actor in cancer

Abstract Cancer is a disease with a significant health impact: approximately 10 million cancer-related deaths occur worldwide every year. Cancer research has unraveled the diverse nature between different cancer types, patients and even within a patient or an individual tumor. Understanding the variating biology is the key in creating biomarkers to detect cancer and discover new targets for cancer drug treatment. Taking advantage of the new biomedical tools for cancer research will help to improve the efficiency and thoroughness of cancer research. Matrix metalloproteinases (MMP) are proteolytic enzymes that degrade the components of extracellular matrix, but also participate in, for example, cell signalling by processing cytokines. Many different substrates for different MMPs have been described, but we lack a complete picture of their substrate repertoires and thus their effects in healthy tissue or pathological conditions. Here, we focus on one of the MMP family members, matrix metalloproteinase 8 (MMP8), and its role in cancer, especially in oral tongue squamous cell carcinoma (OTSCC). The effect of MMP8 on cancer biology and usability depends on the cancer type, as evidenced by our systematic review. By utilizing Biobank Borealis, we showed that possible delays in sample logistics and processing lead to the degradation of blood samples and changes in MMP8 levels, highlighting the importance of well-documented sample handling. In vitro, MMP8 overexpression was shown to decrease the motility and increase cell-cell adhesion of OTSCC cells. Using terminal amine isotopic labeling of substrates (TAILS), we identified 36 novel, potential substrates of MMP8 including FXYD domain-containing ion transport regulator 5 (FXYD5). Cleavage of FXYD5, an anti-adhesive protein upregulated in many cancers, could explain the adhesion-supporting and thus motility-diminishing functions of MMP8 in OTSCC. In all, utilization of various cancer research tools enabled us to paint a fuller picture of “MMP8 in cancer”.Tiivistelmä Syöpä on merkittävien terveysvaikutusten sairaus: noin 10 miljoonaa syöpäkuolemaa tapahtuu vuosittain maailmanlaajuisesti. Syöpätutkimus on paljastanut eroja niin syöpätyyppien ja potilaiden välillä, kuin potilaiden ja yksittäisen kasvaimen sisällä. Muuntelevan syöpäbiologian ymmärtäminen auttaa meitä kehittämään biologisia merkkiaineita syövän tunnistamiseksi sekä löytämään uusia kohdemolekyylejä lääkehoidolle. Uusien biolääketieteellisten menetelmien hyödyntäminen parantaa syöpätutkimuksen tehokkuutta ja perusteellisuutta. Matriksin metalloproteinaasit (MMP) ovat proteolyyttisiä entsyymejä, jotka pilkkovat soluväliaineen rakenneosia, mutta ne osallistuvat myös esimerkiksi solujen väliseen kommunikaatioon sytokiinejä prosessoimalla. Eri MMP:n substraatteja on tunnistettu, mutta meiltä puuttuu hyvä kokonaiskuva monen MMP:n substraattikokonaisuuksista ja siten niiden vaikutuksista terveessä kudoksessa ja sairaustiloissa. Tässä tutkimuksessa keskitymme matriksi metalloproteinaasi 8:n (MMP8) toimintaan syövässä, erityisesti kielisyövässä. MMP8:n syöpäbiologiset vaikutukset ja käytettävyys riippuvat syöpätyypistä, kuten systemaattinen katsauksemme paljasti. Biobankki Borealista hyödyntämällä totesimme verinäytteiden mahdollisten logistiikka- ja prosessointiviiveiden vaikuttavan niiden hajoamiseen ja MMP8-tasoihin, mikä korostaa hyvin dokumentoidun näytekäsittelyn merkitystä. In vitro, MMP8:n yliekspressio vähensi kielisyöpäsolujen liikkuvuutta ja lisäsi solu-solu-adheesiota. Käyttämällä massapektometriaan pohjautuvaa TAILS-menetelmää, tunnistimme 36 uutta MMP8:n substraattia, mukaan lukien ionien siirtoa säätelevän FXYD5-proteiinin. FXYD5:n, joka yli-ilmentyy ja vähentää adheesiota useissa syövissä, pilkonta selittänee MMP8:n adheesiota lisääviä ja liikettä vähentäviä vaikutuksia kielisyövässä. Kokonaisuudessaan erilaisten syöpätutkimuksen työkalujen hyödyntäminen mahdollisti syvemmän ymmärryksen ”MMP8:n roolista syövissä”

Prognostic value of dysadherin in cancer:a systematic review and meta-analysis

Abstract Cancer is a leading cause of death worldwide and novel prognostic factors are reported with increasing numbers. Systematic reviews and meta-analyses on cumulative research data are crucial in estimating the true prognostic value of proposed factors. Dysadherin (FXYD Domain Containing Ion Transport Regulator 5; FXYD5) is a cell membrane glycoprotein that modulates Na+, K+-ATPase activity and cell-cell adhesion. It is abundantly expressed in a variety of cancer cells, but only in a limited number of normal cells and its levels are increased in many different tumor types. The expression or level of dysadherin has been suggested as an independent predictor for metastasis and poor prognosis by number of studies, yet we lack a definitive answer. In this study, we systematically evaluated the prognostic value of dysadherin in cancer and summarized the current knowledge on the subject. PubMed, Scopus, Web of Science and relevant clinical trial and preprint databases were searched for relevant publications and PRISMA and REMARK guidelines were applied in the process. After a careful review, a total of 23 original research articles were included. In each study, dysadherin was pointed as a marker for poor prognosis. Meta-analyses revealed 3- and 1.5-fold increases in the risk of death (fixed effects HR 3.08, 95% CI 1.88–5.06, RR 1.47, 95% CI 1.06–2.05 on overall survival, respectively) for patients with high (>50%) tumoral FXYD5 level. In many studies, a connection between dysadherin expression or level and metastatic behavior of the cancer as well as inverse correlation with E-cadherin level were reported. Thus, we conclude that dysadherin might be a useful prognostic biomarker in the assessment of disease survival of patients with solid tumors

The interplay of matrix metalloproteinase-8, transforming growth factor-β1 and vascular endothelial growth factor-C cooperatively contributes to the aggressiveness of oral tongue squamous cell carcinoma

Matrix metalloproteinase-8 (MMP-8) has oncosuppressive properties in various cancers. We attempted to assess MMP-8 function in oral tongue squamous cell carcinoma (OTSCC). MMP-8 overexpressing OTSCC cells were used to study the effect of MMP-8 on proliferation, apoptosis, migration, invasion and gene and protein expression. Moreover, MMP-8 functions were assessed in the orthotopic mouse tongue cancer model and by immunohistochemistry in patient samples. MMP-8 reduced the invasion and migration of OTSCC cells and decreased the expression of MMP-1, cathepsin-K and vascular endothelial growth factor-C (VEGF-C). VEGF-C was induced by transforming growth factor-β1 (TGF-β1) in control cells, but not in MMP-8 overexpressing cells. In human OTSCC samples, low MMP-8 in combination with high VEGF-C was an independent predictor of poor cancer-specific survival. TGF-β1 treatment also restored the migration of MMP-8 overexpressing cells to the level of control cells. In mouse tongue cancer, MMP-8 did not inhibit metastasis, possibly because it was eliminated in the peripheral carcinoma cells. The suppressive effects of MMP-8 in OTSCC may be mediated through interference of TGF-β1 and VEGF-C function and altered proteinase expression. Together, low MMP-8 and high VEGF-C expression have strong independent prognostic value in OTSCC.1171007101

Novel human lymph node-derived matrix supports the adhesion of metastatic oral carcinoma cells

Abstract Background: 3D culture is increasingly used in cancer research, as it allows the growth of cells in an environment that mimics in vivo conditions. Metastases are the primary cause of morbidity and mortality in cancer patients, and solid tumour metastases are mostly located in lymph nodes. Currently, there are no techniques that model the pre-metastatic lymph node microenvironment in vitro. In this study, we prepared a novel extracellular matrix, Lymphogel, which is derived from lymph nodes, mimicking the tumour microenvironment (TME) of metastatic carcinoma cells. We tested the suitability of the new matrix in various functional experiments and compared the results with those obtained using existing matrices. Methods: We used both commercial and patient-derived primary and metastatic oral tongue squamous cell carcinoma (OTSCC) cell lines. We characterized the functional differences of these cells using three different matrices (human uterine leiomyoma-derived Myogel, human pre-metastatic neck lymph node-derived Lymphogel (h-LG), porcine normal neck lymph node-derived Lymphogel (p-LG) in proliferation, adhesion, migration and invasion assays. We also performed proteomic analyses to compare the different matrices in relation to their functional properties. Results: OTSCC cells exhibited different adhesion and invasion patterns depending on the matrix. Metastatic cell lines showed improved ability to adhere to h-LG, but the effects of the matrices on cell invasion fluctuated non-significantly between the cell lines. Proteomic analyses showed that the protein composition between matrices was highly variable; Myogel contained 618, p-LG 1823 and h-LG 1520 different proteins. The comparison of all three matrices revealed only 120 common proteins. Analysis of cellular pathways and processes associated with proteomes of each matrix revealed similarities of Myogel with h-LG but less with p-LG. Similarly, p-LG contained the least adhesion-related proteins compared with Myogel and h-LG. The highest number of unique adhesion-related proteins was present in h-LG. Conclusions: We demonstrated that human pre-metastatic neck lymph node-derived matrix is suitable for studying metastatic OTSCC cells. As a whole-protein extract, h-LG provides new opportunities for in vitro carcinoma cell culture experiments