Levels Of Tenascin-C Expression Research Articles

IL-33/ST2 axis promotes glioblastoma cell invasion by accumulating tenascin-C

Tenascin-C (TNC), a very large multimeric glycoprotein, is overexpressed in human glioblastomas, leading to a highly motile and invasive phenotype of glioma cells. However, the regulation of TNC expression in glioma has remained unclear until now. Our data suggest that interleukin-33 (IL-33) may promote the accumulation of TNC protein by autocrine or paracrine modes of action in glioma. In the present study, the expression levels of TNC, IL-33, and ST2 were measured in glioma tissue specimens, and the impact of altered IL-33 expression on TNC was investigated in vitro and in vivo. In contrast with control treatment, IL-33 treatment increased TNC expression, and knockdown of IL-33 attenuated TNC expression in glioma cells. Furthermore, IL-33 induced the activation of nuclear factor κB (NF-κB) and increased the expression of TNC in U251 cells. In addition, blockage of the IL-33-ST2-NFκB pathway resulted in downregulation of TNC production. IL-33 promoted glioma cell invasion by stimulating the secretion of TNC. Similarly, knockdown of TNC inhibited the invasiveness of glioma cells. These findings provide a novel perspective on the role of the IL-33/NF-κB/TNC signalling pathway in supporting cancer progression. Thus, targeting the IL-33/NF-κB/TNC signalling pathway may be a useful therapeutic approach in glioma.

Tenascin-C expression is significantly associated with the progression and prognosis in gastric GISTs

Tenascin-C (TNC), an extracellular matrix glycoprotein, has been implicated in progression of various types of cancer. However, few reports exist on TNC expression in gastrointestinal stromal tumors (GISTs). We here attempted to investigate the expression pattern and prognostic significance of TNC in gastric GISTs. We studied TNC expression in 122 gastric GISTs tissue samples by immunohistochemistry, and examined the correlations of TNC expression with clinicopathological parameters and survival of gastric GISTs. The TNC-high expression was observed in 30 (24.6%) of 122 of gastric GISTs. The high levels of TNC expression in gastric GISTs was significantly associated with tumor size (P < .001), multivisceral resection (P = .006), metastasis at initial diagnosis (P = .006), mitotic count (P = .002) and NIH risk classification (P = .015). The TNC mRNA and protein levels were found to significantly downregulated in tumors without progression compared to those tumors which occurred tumor progression during the follow-up period (P < .05). As for the prognostic analysis, it revealed that tumor size, mitotic count, surgical margins, multivisceral resection, and TNC expression were independent predictors of PFS for gastric GISTs (P < .05). The overexpression of TNC may be as a possible marker for the metastatic potential of gastric GISTs patients.

Abstract 1067: Tenascin C in colorectal cancer stroma is a predictive marker for liver metastasis and is a potent target of miR-198 as identified by microRNA analysis

Abstract Background: Liver metastasis in colorectal cancer (CRC) is still one of the major determinants of survival. Tumor stroma has important roles in the development of colorectal cancer (CRC) metastasis. MicroRNAs (miRNAs) are known to regulate the expression of genes involved in many physiological conditions, including cancer. Previous studies reported important roles of miRNAs in cancer cells in CRC development and metastasis. However, only few studies have focused on miRNA expression in CRC stroma. In this study, we revealed the miRNA expression profiles in tumor stroma by miRNA array analysis using Laser captured microdissection (LCM) to identify miRNAs and their target genes as novel molecular markers involved in CRC liver metastasis. Methods: Tumor stroma was isolated from formalin-fixed, paraffin-embedded tissues of primary CRCs with or without liver metastasis by LCM, and miRNA expression was analyzed using TaqMan miRNA arrays. Results: Hierarchical clustering classified 16 CRCs into two groups according to the existence of synchronous liver metastasis. Combinatory target prediction identified tenascin C (TNC) as a predicted target of miR-198, one of the top 10 miRNAs downregulated in tumor stroma of CRCs with synchronous liver metastasis. Immunohistochemical analysis of TNC in 139 primary CRCs revealed that a high staining intensity was correlated with synchronous liver metastasis (P&amp;lt;0.001). TNC staining intensities in the tumor stroma of 83 CRC liver metastases revealed a higher staining in liver metastases than in primary CRCs independently of preoperative chemotherapy. Univariate and multivariate analyses revealed that the TNC staining intensity was an independent prognostic factor to predict postoperative overall survival (P=0.005; n=139) and liver metastasis-free survival (P=0.001; n=128). To evaluate the biological link between miR-198 and TNC, we first checked expression levels of miR-198 in the 16 samples in miRNA array data, and compared them with TNC staining intensities. The CRC samples expressing miR-198 tended to have lower levels of TNC expression in comparison with those without miR-198 expression, although the difference was not statistically significant (P=0.237). Next, we transfected a miR-198 mimic or a miR-198 inhibitor into SW620 human colon cancer cells and CCD-18Co human colon fibroblast cells and analyzed the expression change of TNC protein by western blotting. The relative protein expression of TNC was suppressed by treatment with the miR-198 mimic and increased by miR-198 inhibitor. Conclusions: Alterations of miRNAs in CRC stroma appear to form a metastasis-permissive environment that can elevate TNC to promote liver metastasis. TNC in primary CRC stroma has the potential to be a novel biomarker to predict postoperative prognosis. Citation Format: Tomohiro Murakami, Hirotoshi Kikuchi, Hisahito Ishimatsu, Sanshiro Kawata, Amane Hirotsu, Tomohiro Matsumoto, Yusuke Ozaki, Yoshihiro Hiramatsu, Kinji Kamiya, Megumi Baba, Satoshi Baba, Masatoshi Kitagawa, Takanori Sakaguchi, Hiroyuki Konno, Hiroya Takeuchi. Tenascin C in colorectal cancer stroma is a predictive marker for liver metastasis and is a potent target of miR-198 as identified by microRNA analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1067.

Tenascin-C is a potential cancer-associated fibroblasts marker and predicts poor prognosis in prostate cancer

Tenascin-C (TNC), as a member of the extracellular matrix (ECM), plays an important role in cancer cell proliferation and migration and tumor invasion in various types of cancer. Here, we attempted to investigate the role of TNC as a prognostic factor in prostate cancer. We studied TNC expression via immunohistochemistry in 145 prostate cancer tissue samples. The clinicopathological relevance of TNC expression was examined, as well as other cancer-associated fibroblasts (CAFs)-related factors. Our results showed that the high levels of TNC expression in prostate cancer stroma was significantly associated with lymph node metastasis (P = 0.024) and clinical stage (P = 0.032). Furthermore, TNC was positively correlated with increased micro-vessel density (MVD) (P = 0.017) and tumor associated macrophage (TAM) population (P = 0.025). In both univariate and multivariate Cox regression analyses, TNC (P < 0.001) was an independent poor prognostic factor for overall survival in prostate cancer patients. Moreover, over-expression of TNC (P < 0.001), SMA (P = 0.042) and vimentin (P = 0.010) were significantly correlated with the lower overall survival. In addition, TNC expression in prostate cancer stroma was significantly associated with FSP1 (P = 0.011), SMA (P = 0.021), and vimentin (P = 0.002). In conclusion, our study revealed that high level of TNC as a potential biomarker of CAFs was significantly correlated with the poor prognosis for prostate cancer patients.

Tenascin-C Downregulates Wnt Inhibitor Dickkopf-1, Promoting Tumorigenesis in a Neuroendocrine Tumor Model

The extracellular matrix molecule tenascin-C (TNC) is a major component of the cancer-specific matrix, and high TNC expression is linked to poor prognosis in several cancers. To provide a comprehensive understanding of TNC's functions in cancer, we established an immune-competent transgenic mouse model of pancreatic β-cell carcinogenesis with varying levels of TNC expression and compared stochastic neuroendocrine tumor formation in abundance or absence of TNC. We show that TNC promotes tumor cell survival, the angiogenic switch, more and leaky vessels, carcinoma progression, and lung micrometastasis. TNC downregulates Dickkopf-1 (DKK1) promoter activity through the blocking of actin stress fiber formation, activates Wnt signaling, and induces Wnt target genes in tumor and endothelial cells. Our results implicate DKK1 downregulation as an important mechanism underlying TNC-enhanced tumor progression through the provision of a proangiogenic tumor microenvironment.

Paper 162: Increased Expression of Tenascin-C in Osteotendinous Healing Junction under Mechanical Strain

The developments of the fibrocartilage in musculoskeletal organs are strongly related the mechanical strain, however, the influence of mechanical strain on fibrocartilage zone regeneration and remodeling during osteotendinous junction repair are currently poorly understood (1Benjamin M. Ralphs J.R. Fibrocartilage in tendon and ligaments: An adaptation to compressive load.J Anat. 1998; 193: 481-494Crossref PubMed Google Scholar). In present study, the expression of tenascin-C and transforming growth factor – ß 1(TGFß1), two key factors involved in strength and elasticity of extracellular matrix remodeling (2Jarvinen T.A. Jozsa L. Kannus P. Jarvinen T.L. Kvist M. Hurme T. Isola J. Kalimo H. Jarvinen M. Mechanical loading regulates tenascin-C expression in the osteotendinous junction.J Cell Sci. 1999; 18: 3157-3166Google Scholar, 4Zhao Y. Young S.L. TGF-beta regulates expression of tenascin alternative-splicing isoforms in fetal rat lung.Am J Physiol. 1995; 268: 173-180Google Scholar), were examined in the healing osteotendinous junction under controlled mechanical loading through quadriceps muscles tension induced by functional electric stimulation (FES) by using our established partial patellectomy model in rabbits (5Wang W. Chen H.H. Yang X.H. Xu G. Chan K.M. Qin L. Postoperative programmed muscle tension augmented osteotendinous junction repair.Int. J Sports Med. 2007; 28: 691-696Crossref PubMed Scopus (14) Google Scholar). Standard transverse osteotomy was performed between the proximal 2/3 and the distal 1/3 of the patella, and distal 1/3 of the patella was excised. The patellar tendon was directly sutured back to the proximal remaining patella while protected with a ‘figure of eight' tension band wire. The knee was immobilized with long leg cast in 90° of knee flexion, 21 rabbits were randomly divided into two groups. The controlled mechanical strain generated from quadriceps muscle contraction induced by FES was applied on the healing patella–patellar tendon junction for 30 minutes per day, 5 days per week for 6 weeks in FES group compaired with non-mechanical loading in control group. The quadriceps muscles-patella-patellar tendon-tibia (QPPT) complex was harvested at the end of postoperative week 6 and 12 respectively for biomechanical testing (n=5) and semiquantitative histology (n=2). The samples after biomechanical testing were also used for histological quantification. Image quantification of fibrocartilage zone and proteoglycan content, semiquantitation of immunohistometry of the expression of tenascin-C and transforming growth factor – ß 1(TGFß1) in the healing osteotendinous junction, ultimate stress of (QPPT) complex were compared by two-way ANOVA between the FES group and control group at each healing time points. Statistical significance level was set at p ? 0.05. As compared with control group, better organized collagen fibers orientated along with mechanical loading axis was observed under polarized microscope to bridge the healing osteotendinous interface at both week 6 and 12. An elevated proteoglycant stainability and larger intermitted fibrocartilage zone were found in the osteotendinous healing interface in FES group compared to control group at both week 6 and 12. Semiquantitation of immunohistometry revealed that the expression levels of Tenascin-C and TGFß-1 increased in the osteotendinous healing interface in FES group compared to control group at both week 6 and 12. The decreased cross sectional area, increased failure load and ultimate strength of all healing osteotendinous interface observed with healing over time, but more significant in FES group as compared to the controls at each healing time point. However, the ultimate strength of FES group and control group only reached to 20.2±3.6% and 11.6±4.5% of the intact contralateral hind limb respectively. The failure mode of the tested healing osteotendinous complex specimens in control group was found at the initial osteotomy site, but at the sub-fibrocartilage woven bone in FES group. The dramatically increased levels of tenascin C and TGF-ß1 expression in the healing osteotendinous junction induced by mechanical strain generated from muscle tension contraction have not been reported before. The much better restoration of fibrocartilage zone in terms of morphology as well as biomechanical properties under the mechanical strain was also shown. The present study indicated that the mechanical strain has beneficial effects on osteotendinous healing processes by influencing the extracellular matrix remodeling at fibrocartilage zone. The dilemma between prolonged immobilization in order to providing a static environment for facilitating osteotendinous junction repair and the need of early functional rehabilitation remains a challenge in surgical osteotendinous junction repair clinically (3Thomopoulos S. Williams G.R. Soslowsky L.J. Tendon to bone healing: differences in biomechanical, structural, and compositional properties due to a range of activity levels.J Biomech Eng. 2003; 125: 106-113Crossref PubMed Scopus (296) Google Scholar). Findings of our study suggest that the early muscle tension for patients underwent osteotendinous junction repair should be conducted.

Increased Tenascin C And Toll-Like Receptor 4 Levels in Visceral Adipose Tissue as a Link between Inflammation and Extracellular Matrix Remodeling in Obesity

Obesity is associated with an altered inflammatory and extracellular matrix (ECM) profile. Tenascin C (TNC) is an ECM glycoprotein with proinflammatory effects. We aimed to explore the expression levels of TNC in adipose tissue analyzing the contribution of adipocytes and stromovascular fraction cells (SVFC) as well as its impact on inflammation and ECM regulation. We also analyzed the effect of the stimulation with TNF-α and lipopolysaccharide (LPS) on both SVFC and adipocytes. Samples obtained from 75 subjects were used in the study. Expression levels of TNC, TLR4, MMP2, and MMP9 were analyzed in visceral adipose tissue (VAT) as well as in both adipocytes and SVFC. In addition, Tnc expression was measured in two mice models of obesity. We show, for the first time, that VAT expression levels of TNC are increased in normoglycemic and type 2 diabetic obese patients (P<0.01) as well as in obese patients with nonalcoholic steatohepatitis (P<0.01). Furthermore, expression levels of Tnc in epididymal adipose tissue from two different mice models of obesity were significantly increased (P<0.01). TNC and TLR4 were mainly expressed by SVFC, and its expression was significantly enhanced (P<0.01) by TNF-α treatment. LPS treatment also increased mRNA levels of TNC. Moreover, the addition of exogenous TNC induced (P<0.05) TLR4 and CCL2 mRNA expression in human adipocyte cultures. These findings indicate that TNC is involved in the etiopathology of obesity via visceral adipose tissue inflammation representing a link with ECM remodeling.

Abstract 485: Epidermal growth factor-like repeats of Tenascin C promote melanoma cell invasion, possibly by enabling mesenchymal to ameboid migration transition

Abstract In malignant melanoma, the most dangerous skin cancer, Tenascin-C (TNC) is overexpressed, especially at the invasive fronts of invasive melanomas. We found that the migration and invasiveness of a set of melanoma cell lines correlates with levels of TNC expression suggesting, surprisingly, that the melanoma cells themselves provide much of this matrix component. Adding TNC to matrices enhanced individual cell migration in cells that did not express much TNC, whereas it did not enhance the rapid motility of the cells expressing high levels. As TNC is a multidomain protein, the signaling pathways that promote this motogenic invasiveness are not obvious. Herein, we examined the potential role of the EGF-like repeats of TNC that have been shown to preferentially activate motogenic-signaling cascades. To analyze the effects of EGFL signaling on melanoma migration and invasion, we overexpressed TNC fragment containing the assembly and EGFL domains of TNC (TNCEGFL). Interestingly, this led to impaired cell migration in in vitro wound healing assays and live cell tracking experiments. This was likely due to a shift in the adhesion-contractility balance during cell migration. TNCEGFL expressing cells presented a spindle-shaped morphology and formed cords in culture. Expression of TNCEGFL delayed melanoma cell attachment and spreading, as determined in cell adhesion and inverted centrifugation assays. This anti-adhesive phenotype was concomitant with increased MLC2 phosphorylation. Inhibition of ROCK activity, which drives transcellular contractility, restored adhesion of TNCEGFL expressing cells and improved migration in 2D. In distinction to the findings in 2D, TNCEGFL expressing cells had higher invasion potential in Matrigel invasion assays, with cells expressing high TNC having rounded “ameboid” morphology, underlying the importance of morphological shifts, and possibly ROCK signaling, in 3D movement through barrier matrices during invasion and dissemination. These results imply that TNCEGFL repeats are involved in melanoma invasion both by directly influencing cell migration and indirectly changing the tumor microenvironment composition, possibly allowing transition from mesenchymal to ameboid migrational mode. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 485. doi:1538-7445.AM2012-485

Association of invasion-promoting tenascin-C additional domains with breast cancers in young women

IntroductionTenascin-C (TNC) is a large extracellular matrix glycoprotein that shows prominent stromal expression in many solid tumours. The profile of isoforms expressed differs between cancers and normal breast, with the two additional domains AD1 and AD2 considered to be tumour associated. The aim of the present study was to investigate expression of AD1 and AD2 in normal, benign and malignant breast tissue to determine their relationship with tumour characteristics and to perform in vitro functional assays to investigate the role of AD1 in tumour cell invasion and growth.MethodsExpression of AD1 and AD2 was related to hypoxanthine phosphoribosyltransferase 1 as a housekeeping gene in breast tissue using quantitative RT-PCR, and the results were related to clinicopathological features of the tumours. Constructs overexpressing an AD1-containing isoform (TNC-14/AD1/16) were transiently transfected into breast carcinoma cell lines (MCF-7, T-47 D, ZR-75-1, MDA-MB-231 and GI-101) to assess the effect in vitro on invasion and growth. Statistical analysis was performed using a nonparametric Mann-Whitney test for comparison of clinicopathological features with levels of TNC expression and using Jonckheere-Terpstra trend analysis for association of expression with tumour grade.ResultsQuantitative RT-PCR detected AD1 and AD2 mRNA expression in 34.9% and 23.1% of 134 invasive breast carcinomas, respectively. AD1 mRNA was localised by in situ hybridisation to tumour epithelial cells, and more predominantly to myoepithelium around associated normal breast ducts. Although not tumour specific, AD1 and AD2 expression was significantly more frequent in carcinomas in younger women (age ≤40 years; P < 0.001) and AD1 expression was also associated with oestrogen receptor-negative and grade 3 tumours (P < 0.05). AD1 was found to be incorporated into a tumour-specific isoform, not detected in normal tissues. Overexpression of the TNC-14/AD1/16 isoform significantly enhanced tumour cell invasion (P < 0.01) and growth (P < 0.01) over base levels.ConclusionsTogether these data suggest a highly significant association between AD-containing TNC isoforms and breast cancers in younger women (age ≤40 years), which may have important functional significance in vivo.

Differential expression of tenascin-C in the developing human lung: an immunohistochemical study

Much of the specification for the basic embryonic body plan is the result of a hierarchy of developmental decisions at different developmental times. The extracellular matrix (ECM) appears to be a very dynamic structure during embryogenesis. One of the mesenchymal ECM proteins, tenascin, is reported to be transiently expressed during embryonic tissue development, and is absent or much reduced in most fully developed organs. The respiratory system is an outgrowth of the ventral wall of the foregut, and the epithelium of the larynx, trachea, bronchi and alveoli is of endodermal origin. The cartilaginous and muscular components are of mesodermal origin. The aim of this study was to investigate the role of tenascin-C (TNC) in the developing human lung, during the pseudoglandular, canalicular and saccular stage of lung maturation. Formalin-fixed, paraffin-embedded tissue from the lungs of 30 embryos (10 corresponding to the 10th to the 16th gestational week (pseudoglandular stage), 10 to the 17th to the 23rd gestational week (canalicular stage), and 10 to the 24th to the 27th gestational week (saccular stage), were investigated by conventional histology and immunohistology for the expression levels of TNC. The changes observed in the distribution patterns suggest that during embryogenesis, the rate of tenascin synthesis changes significantly. During the pseudoglandular stage, the density of cells expressing TNC was higher in the condensing mesenchyme surrounding the epithelial glands than in the epithelial cells, whereas the inverse result was observed during the canalicular stage. During the saccular stage the pattern of immunoreactivity with TNC was lower than those of the pseudoglandular and canalicular stage, either in epithelial or mesenchymal cells, but it was highly expressed in the basement membranes. This restricted spatiotemporal distribution suggests that tenascin has a key role (1) in mesenchymal tissue remodeling during the pseudoglandular stage, a period that describes the development of the complete bronchial tree and (2) on the epithelial cell shape and function during the canalicular stage, a period that describes the formation of pneumocytes type I and pneumocytes type II. The later, will produce the surfactant, a phospholipid-rich fluid capable of lowering surface tension at the air-alveolar interface. During the saccular stage, tenascin was present mainly in the basement membranes surrounding the acinar and vascular structures, indicating a supporting and mechanical role.

Upregulation of Tenascin-C Expression by IL-13 in Human Dermal Fibroblasts via the Phosphoinositide 3-kinase/Akt and the Protein Kinase C Signaling Pathways

In this study, we examined the genes targeted by IL-13 in human dermal fibroblasts using a cDNA microarray. We focused on the tenascin-C (TN-C) gene, which was identified as one of the genes induced by IL-13. IL-13 induced transcriptional activity of TN-C. IL-13-mediated TN-C expression was inhibited by treatment with wortmannin or LY294002, or Calphostin C. IL-13 induced the phosphorylation of the phosphoinositide 3-kinase (PI3K) regulatory subunit p85, induced tyrosine phosphorylation of Akt, upregulated Akt kinase activity, and activated protein kinase C (PKC)-delta and -epsilon. The IL-13-induced increase in TN-C protein expression was abrogated by the transfection of a dominant-negative mutant of Akt, PKC-delta, or PKC-epsilon. In conclusion, we showed that the PI3K/Akt and/or PKC signaling pathways are essential for the IL-13-mediated increase in TN-C. Both serum levels of IL-13 and the expression levels of TN-C in the dermis are increased in patients with systemic sclerosis. Our findings suggest that the expression of TN-C is upregulated in this disease due to IL-13 signaling, and that a blockade of the PI3K or PKC signaling pathway may also have therapeutic value by reducing the amount of TN-C produced during fibrosis.

Retinoic acid and 1,25-dihydroxyvitamin D3 inhibit tenascin-C expression in rat glioma C6 cells.

Tenascin-C (Tn-C) is an extracellular matrix protein with growth-, invasive-, and angiogenesis-promoting activities. Tn-C is upregulated during wound healing, tumorigenesis, and other pathological conditions. Highly malignant gliomas with poor prognosis exhibit high levels of Tn-C expression. Here we demonstrate that Tn-C RNA expression in glioma C6 cells is inhibited in a dose-dependent manner by retinoic acid (RA) and 1,25-dihydroxyvitamin D3 (1,25-D3). No additive or synergistic effects were found. Inhibition is maximum 24 hr after RA or 1,25-D3 treatment, prior to a delayed cytotoxic effect starting at day 4-5 of treatment, and correlates with a reduction in the synthesis of Tn-C protein. Tn-C expression is also inhibited, but to a lesser extent by prostaglandin D2 (PGD2). Furthermore, both RA and 1,25-D3, but not PGD2 abolish the induction of Tn-C by the tumor promoter 12-O-tetradecanoyl phorbol 13-acetate. The inhibition of Tn-C expression might be relevant for the anti-cancer activity of RA and 1,25-D3.