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Asporin is predominantly expressed in CAFs of gastric cancer. (A) Left: total cell lysates were prepared from CAFs or NFs (established from three patients with scirrhous gastric cancer) and from 44As3 cells. Lysates were subjected to IB with anti-Asporin and anti-Decorin antibodies. Right: total RNA was isolated from each cell type and used for RT–PCR detection of mRNAs encoding Biglycan, Tsukushi and α-SMA. (B) Left: NF-37 (1.0 × 106) and 44As3 cells (2.0 × 106) were subcutaneously implanted into nude mice, separately or together, for 5 days. Right: NF-37 cells were cultured in CM of 44As3 cells for 4 days. Lysates were prepared from the subcutaneous nodules or cells, respectively, and immunoblotted with anti-Asporin antibody. (C) Asporin expression in human scirrhous gastric cancer tissue. Immunohistochemistry with anti-Asporin antibody. Enlargement of square a is shown at right. Ca indicates the cancer cell. Bottom: the same specimen was immunostained with anti-Asporin (red), anti-α-smooth muscle actin (green) and DAPI (blue). Regions corresponding to squares a, b and c are shown. M, mucosa; MP, muscularis propria; SM, submucosa. (a–c) Bar, 50 μm. Bar in the large picture indicates 200 μm.
Source publication
Scirrhous gastric cancer, which has the worst prognosis among the various types of gastric cancer, is highly invasive and associated with abundant stromal fibroblasts. Although cancer-associated fibroblasts (CAFs) have been proposed to generate a tumor-supportive extracellular matrix that promotes the expansion of this type of cancer, the molecular...
Contexts in source publication
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... is a secretory protein belonging to the SLRP family of ECM proteins. Western blots revealed that Asporin was expressed at higher levels in all CAF cell lines than in all NF cells that we examined, whereas it was not expressed in 44As3 scirrhous gastric cancer cell line ( Figure 1A). We also analyzed the expression of other SLRP-family members, namely, Decorin and Biglycan, which like Asporin are members of the canonical class I of SLRPs, and Tsukushi, a member of the typical class IV. ...
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... also analyzed the expression of other SLRP-family members, namely, Decorin and Biglycan, which like Asporin are members of the canonical class I of SLRPs, and Tsukushi, a member of the typical class IV. Expression levels of these proteins varied among cell lines, and no significant difference between CAFs and NFs was revealed by immunoblot- ting (IB) or RT-PCR ( Figure 1A). Information regarding other members of the SLRP family, obtained by microarray analyses, is shown in Table 1. ...
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... expression of Asporin was significantly elevated in CAFs, we then investigated whether gastric cancer cells induce Asporin expression in fibroblasts. When NF-37 and 44As3 cells were injected subcutaneously into nude mice, either together or separately, Asporin expression was induced in tumor nodules consisting of NFs and 44As3 5 days after implantation, whereas no expression was detected in nodules containing NFs or 44As3 alone ( Figure 1B, left). In addition, expression of Asporin in NF-37 was elevated upon incubation of the cells in conditioned medium (CM) from 44As3 cells, although the level of expression was lower than in CAFs ( Figure 1B, right). ...
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... NF-37 and 44As3 cells were injected subcutaneously into nude mice, either together or separately, Asporin expression was induced in tumor nodules consisting of NFs and 44As3 5 days after implantation, whereas no expression was detected in nodules containing NFs or 44As3 alone ( Figure 1B, left). In addition, expression of Asporin in NF-37 was elevated upon incubation of the cells in conditioned medium (CM) from 44As3 cells, although the level of expression was lower than in CAFs ( Figure 1B, right). ...
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... we examined expression of Asporin in human gastric cancer tissues, using immunohistochemistry. Fibrillar or amorphous staining of Asporin was detected in cancer stroma, whereas it was not observed in the normal mucosa in all cases that we examined (n ¼ 10) ( Figure 1C). When Asporin and cytokeratin 20, a marker of many types of epithelial cells, were immunostained in human gastric cancer tissue, no Asporin deposition was detected in nests of cancer cells (Supplementary Figure 1a). ...
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... we examined expression of Asporin in human gastric cancer tissues, using immunohistochemistry. Fibrillar or amorphous staining of Asporin was detected in cancer stroma, whereas it was not observed in the normal mucosa in all cases that we examined (n ¼ 10) ( Figure 1C). When Asporin and cytokeratin 20, a marker of many types of epithelial cells, were immunostained in human gastric cancer tissue, no Asporin deposition was detected in nests of cancer cells (Supplementary Figure 1a). Instead, the staining pattern of Asporin matched that of Vimentin-positive fibroblasts (Supplementary Figure 1b). ...
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... Asporin and cytokeratin 20, a marker of many types of epithelial cells, were immunostained in human gastric cancer tissue, no Asporin deposition was detected in nests of cancer cells (Supplementary Figure 1a). Instead, the staining pattern of Asporin matched that of Vimentin-positive fibroblasts (Supplementary Figure 1b). High expression of Asporin was also observed in CAFs from early-stage gastric cancers, when cancer invasion is restricted to the submucosal space (n ¼ 5; data not shown). ...
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... CAFs, a-SMA-positive cells are considered to represent a biologically active myofibroblast-like population. Within tumors, Asporin was diffusely stained in the central region of submucosal invasion, where a-SMA-positive cells also accumu- lated ( Figure 1C, bottom a). Asporin deposits at least beneath and surround the a-SMA-positive CAFs (Supplementary Figure 1c). ...
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... tumors, Asporin was diffusely stained in the central region of submucosal invasion, where a-SMA-positive cells also accumu- lated ( Figure 1C, bottom a). Asporin deposits at least beneath and surround the a-SMA-positive CAFs (Supplementary Figure 1c). In the periphery of tumors, a-SMA-positive cells were less abundant, although diffuse deposition of Asporin persisted ( Figure 1C, bottom b). ...
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... deposits at least beneath and surround the a-SMA-positive CAFs (Supplementary Figure 1c). In the periphery of tumors, a-SMA-positive cells were less abundant, although diffuse deposition of Asporin persisted ( Figure 1C, bottom b). We detected neither Asporin nor a-SMA in the normal submucosal area, except for the expected a-SMA staining in the muscle layer of the arterial wall ( Figure 1C, bottom c). ...
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... the periphery of tumors, a-SMA-positive cells were less abundant, although diffuse deposition of Asporin persisted ( Figure 1C, bottom b). We detected neither Asporin nor a-SMA in the normal submucosal area, except for the expected a-SMA staining in the muscle layer of the arterial wall ( Figure 1C, bottom c). These observations indicate that Asporin is broadly expressed in a- SMA-positive and negative CAFs throughout the entire tumor. ...
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Decorin (DCN) is an important small leucine-rich proteoglycan present in the extracellular matrix (ECM) of many organs and tissues. Endothelial progenitor cells (EPCs) are able to interact with the surrounding ECM and bind to molecules such as DCN. Here, we recombinantly produced full-length human DCN under good laboratory practice (GLP) conditions...
Citations
... Indeed, asporin competes with decorin in collagen binding and promotes osteoblast collagen mineralization [49]. Due to its ubiquitous expression, asporin has been implicated as an activator of invasion in schirrhous gastric carcinomas [50], as well as growth and migration of gastric cancers cells [51,52]. Moreover, stromalderived asporin is a biomarker associated with prostate cancer progression [53], bladder cancer where asporin levels correlate with the degree of malignancy [54]. ...
Solid tumors present a formidable challenge in oncology, necessitating innovative approaches to improve therapeutic outcomes. Proteoglycans, multifaceted molecules within the tumor microenvironment, have garnered attention due to their diverse roles in cancer progression. Their unique ability to interact with specific membrane receptors, growth factors, and cytokines provides a promising avenue for the development of recombinant proteoglycan‐based therapies that could enhance the precision and efficacy of cancer treatment. In this study, we performed a comprehensive analysis of the proteoglycan gene landscape in human breast carcinomas. Leveraging the available wealth of genomic and clinical data regarding gene expression in breast carcinoma and using a machine learning model, we identified a unique gene expression signature composed of five proteoglycans differentially modulated in the tumor tissue: Syndecan‐1 and asporin (upregulated) and decorin, PRELP and podocan (downregulated). Additional query of the breast carcinoma data revealed that serglycin, previously shown to be increased in breast carcinoma patients and mouse models and to correlate with a poor prognosis, was indeed decreased in the vast majority of breast cancer patients and its levels inversely correlated with tumor progression and invasion. This proteoglycan gene signature could provide novel diagnostic capabilities in breast cancer biology and highlights the need for further utilization of publicly available datasets for the clinical validation of preclinical experimental results.
... ASPN, also known as periodontal ligament associated protein-1 (PLAP-1) is a member of class I small leucinerich proteoglycans (SLRP) class I family, which directly binds to type I collagen and can play a crucial part in collagen fibrillogenesis [60]. In various tumors, ASPN, expressed primarily by stromal fibroblasts, is positively associated with pancreatic, colorectal, gastric, and prostate cancers [43,[61][62][63]. While in breast cancer, also plays dual roles with both pro-and anti-tumor effects [64]. ...
Background
Heart failure (HF) and cancer share common risk factors and pathophysiological mechanisms, including fibrosis. Identifying biomarkers and therapeutic targets for both conditions is crucial.
Materials and methods
RNA sequencing data from HF patients were analyzed to identify 12 genes associated with myocardial fibrosis. Validation was performed using public datasets, and functional enrichment analyses were conducted. Gene expression patterns and prognostic value in various cancers were assessed.
Results
Fibromodulin (FMOD), Periostin (POSTN), Latent Transforming Growth Factor Beta Binding Protein 2 (LTBP2), Collagen Type I Alpha 1 Chain (COL1A1), Collagen Type VIII Alpha 1 Chain (COL8A1), Asporin (ASPN), and Hemoglobin Subunit Beta (HBB) showed significant dysregulation in heart failure tissues and were implicated in multiple cancer types. Pan-cancer analysis revealed associations between these genes and prognosis. Correlations with cancer-associated fibroblasts were also observed.
Conclusion
FMOD, POSTN, LTBP2, COL1A1, COL8A1, ASPN, and HBB are potential biomarkers for HF and cancer with fibrotic microenvironments. Targeting fibrosis may offer novel therapeutic approaches. Further validation and mechanistic studies are needed. This study contributes to understanding HF and cancer at the molecular level and suggests personalized treatment strategies.
... The preceding infiltration of CAFs followed by cancer cells is reported in squamous cell carcinoma, lung adenocarcinoma and gastric cancer (Gaggioli et al. 2007;Neri et al. 2015;Satoyoshi et al. 2015). This mode of invasion does not depend on the A c c e p t e d M a n u s c r i p t 6 invasive properties of the cancer cells themselves, and it has been suggested that the invasive properties of CAFs contribute to cancer cell invasion and are ratelimiting factors (Gaggioli et al. 2007;Neri et al. 2015). ...
... They also found that CAFs infiltrated gels even when CAFs were cultured alone (Gaggioli et al. 2007). It has also been reported that CAFs are present at the leading edge of infiltration and induce cancer cell invasion in lung and gastric cancers (Neri et al. 2015;Satoyoshi et al. 2015). If a similar invasion pattern occurs in gallbladder cancer, the In addition to the mechanism of "CAF-led invasion", in which CAF itself precedes invasion, the secreted proteins from CAF enhance the malignancy of cancer cells (Yamamura et al. 2015). ...
... Direct cancer cells-CAFs interaction also enhances the invasion of lung carcinoma cells in a 3D co-culture system (Otomo et al., 2014) and in vivo (Duda et al., 2010). Gastric carcinoma studies showed that CAFs promote a strong invasive phenotype when in direct contact with scirrhous gastric carcinoma cells (Yamaguchi et al., 2014;Satoyoshi et al., 2015), whereas CAFs conditioned media did not, emphasizing the importance of direct contact between cancer cells and fibroblasts in invasion (Satoyoshi et al., 2015). Furthermore, TGFβ1 enhances the dissemination of colon carcinoma cells to the liver through attachment to CAFs (Gonzalez-Zubeldia et al., 2015). ...
... Direct cancer cells-CAFs interaction also enhances the invasion of lung carcinoma cells in a 3D co-culture system (Otomo et al., 2014) and in vivo (Duda et al., 2010). Gastric carcinoma studies showed that CAFs promote a strong invasive phenotype when in direct contact with scirrhous gastric carcinoma cells (Yamaguchi et al., 2014;Satoyoshi et al., 2015), whereas CAFs conditioned media did not, emphasizing the importance of direct contact between cancer cells and fibroblasts in invasion (Satoyoshi et al., 2015). Furthermore, TGFβ1 enhances the dissemination of colon carcinoma cells to the liver through attachment to CAFs (Gonzalez-Zubeldia et al., 2015). ...
The interaction between the tumor microenvironment (TME) and the cancer cells is a complex and mutually beneficial system that leads to rapid cancer cells proliferation, metastasis, and resistance to therapy. It is now recognized that cancer cells are not isolated, and tumor progression is governed among others, by many components of the TME. The reciprocal cross-talk between cancer cells and their microenvironment can be indirect through the secretion of extracellular matrix (ECM) proteins and paracrine signaling through exosomes, cytokines, and growth factors, or direct by cell-to-cell contact mediated by cell surface receptors and adhesion molecules. Among TME components, cancer-associated fibroblasts (CAFs) are of unique interest. As one of the most abundant components of the TME, CAFs play key roles in the reorganization of the extracellular matrix, facilitating metastasis and chemotherapy evasion. Both direct and indirect roles have been described for CAFs in modulating tumor progression. In this review, we focus on recent advances in understanding the role of direct contact between cancer cells and cancer-associated fibroblasts (CAFs) in driving tumor development and metastasis. We also summarize recent findings on the role of direct contact between cancer cells and CAFs in chemotherapy resistance.
... ASPN plays a role in the feedback mechanism with TGF-β1 in knee osteoarthritis, and it is involved in its pathogenesis [19]. However, in recent years, ASPN has been reported to be associated with various cancers, such as breast [20], gastric [21], and colorectal cancers [22]. ...
Nonalcoholic fatty liver disease (NAFLD) develops as a result of unhealthy lifestyle but improves with laparoscopic sleeve gastrectomy (LSG). The transforming growth factor (TGF)-β signaling pathway reportedly contributes to liver fibrosis, mainly in animal experiments. The aim of the present study was to evaluate changes in serum proteins before and after LSG by proteomic analysis and to investigate their association with NAFLD. This study enrolled 36 severely obese patients who underwent LSG at our hospital from January 2020 to April 2022. As a pilot study, proteomic analysis was conducted on six patients using serum collected before and at 6 months after LSG, and significantly fluctuating proteins were extracted. Subsequently, verification by enzyme-linked immunosorbent assay (ELISA) using collected serum was performed on the remaining 30 patients. The mean weight of enrolled patients was 118.5 kg. Proteomic analysis identified 1,912 proteins, many of which were related to the TGF-β signaling pathway. Among these proteins, we focused on five TGF-β-related proteins: asporin, EMILIN-1, platelet factor-4, serglycin, and thrombospondin-1. Verification by ELISA revealed that asporin (p = 0.006) and thrombospondin-1 (p = 0.043) levels significantly fluctuated before and after LSG. Univariate analysis with a linear regression model showed that aspartate aminotransferase (p = 0.045), asporin (p = 0.011), and thrombospondin-1 (p = 0.022) levels were significantly associated with postoperative liver fibrosis. On multivariate analysis, asporin was an independent prognostic factor for postoperative liver fibrosis (95% confidence interval: 0.114–1.291, p = 0.002). TGF-β-related proteins dramatically fluctuated before and after LSG and were correlated with NAFLD pathogenesis. Asporin may be a useful prognostic marker of liver fibrosis in NAFLD after LSG.
... Three-dimensional (3D) Matrigel invasion assay. The 3D Matrigel invasion assay was performed as previously described (27)(28)(29)(30). Cancer cells were labeled with DiI (Thermo Fisher Scientific, Inc.). ...
Ras-related protein 25 (Rab25) is a member of small GTPase and is implicated in cancer cell progression of various types of cancer. Growing evidence suggests the context-dependent role of Rab25 in cancer invasiveness. Claudin-7 is a tight junction protein and has been known to suppress cancer cell invasion. Although Rab25 was reported to repress cancer aggressiveness through recycling β1 integrin to the plasma membrane, the detailed underlying mechanism remains to be elucidated. The present study identified the critical role of claudin-7 in Rab25-induced suppression of colon cancer invasion. 3D Matrigel system and modified Boyden chamber analysis showed that enforced expression of Rab25 attenuated colon cancer cell invasion. In addition, Rab25 inactivated epidermal growth factor receptor (EGFR) and increased E-cadherin expression. Unexpectedly, it was observed that Rab25 induces claudin-7 expression through protein stabilization. In addition, ectopic claudin-7 expression reduced EGFR activity and Snail expression as well as colon cancer cell invasion. However, silencing of claudin-7 expression reversed the tumor suppressive role of Rab25, thereby increasing colon cancer cell invasiveness. Collectively, the present data indicated that Rab25 inactivates EGFR and colon cancer cell invasion by upregulating claudin-7 expression.
... Similar to NID2, asporin has a stabilizing function in binding to ECM collagens [41]. Up-regulation of asporin has been observed in colorectal, gastric, pancreatic, and prostate cancer [42][43][44][45]. The most important signaling pathways associated with the dysregulation of asporin expression are transforming growth factor beta, epidermal growth factor receptor, and CD44 signaling leading to the promotion of cell proliferation, migration, and invasiveness [46]. ...
Vestibular schwannoma is the most common benign neoplasm of the cerebellopontine angle. Its first symptoms include hearing loss, tinnitus, and vestibular symptoms, followed by cerebellar and brainstem symptoms, along with palsy of the adjacent cranial nerves. However, the clinical picture has unpredictable dynamics and currently, there are no reliable predictors of tumor behavior. Hence, it is desirable to have a fast routine method for analysis of vestibular schwannoma tissues at the molecular level. The major objective of this study was to verify whether a technique using in‐sample specific protein digestion with trypsin would have the potential to provide a proteomic characterization of these pathological tissues. The achieved results showed that the use of this approach with subsequent liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) analysis of released peptides allowed a fast identification of a considerable number of proteins in two differential parts of vestibular schwannoma tissue as well as in tissues of control healthy samples. Furthermore, mathematical analysis of MS data was able to discriminate between pathological vestibular schwannoma tissues and healthy tissues. Thus, in‐sample protein digestion combined with LC‐MS/MS separation and identification of released specific peptides followed by mathematical analysis appears to have the potential for routine characterization of vestibular schwannomas at the molecular level. Data are available via ProteomeXchange with identifier PXD045261.
... Biglycan interacts with CD44, which increases M1 autophagy [222]. Extracellular secreted asporin binds to CD44 to activate Rac1 [223,224]. The GAG chains of biglycan and lumican may interact with CD44, because CD44 interacts with the CS side chain of Serglycin [225]. ...
Small leucine rich repeat proteoglycans (SLRPs) exist in the extracellular matrix. SLRPs contain tandem arrays of LRRs flanked by cysteine clusters at the both N-and C-termini. The extreme N-and/or C-termini contain low complexity sequences, glycosaminoglycan (GAG) chain and/or sulfated tyrosine residues in some members of SLRPs. The LRR solenoid structure may be divided into four parts consisting of a concave surface, an ascending surface, a convex surface, and a descending surface. SLRPs share many biological functions including collagen fibrillogenesis, signaling, innate immunity, and inflammation through the binding of various ligands. We undertake a comprehensive literature search of publications in order to make a list of ligands of SLRPs. We describe and discuss the interacting sites of SLRPs to binding partners. The protein-ligand interactions occur on not only the concave surface but also the ascending surface and the Nor C-terminal capping regions. In addition, the extreme N-and/or C-terminal regions with the GAG chains or sulfated tyrosine residues participate in ligand-interactions.
... 2.9. Three-dimensional (3D) Matrigel invasion assay 3D Matrigel invasion assay was performed as described previously [22][23][24][25]. Cancer cells were labeled with DiI (Thermo sher Scienti c, Waltham, MA, USA). ...
Purpose
Ras-related protein 25 (Rab25) is a member of small GTPase and implicated in various cancer cell progression. Growing evidence suggests the context-dependent roles of Rab25 in cancer invasiveness. Claudin-7 is a tight junction protein and has been known to suppress cancer cell invasion. Although Rab25 was reported to repress cancer aggressiveness through recycling β1 integrin to the plasma membrane, the detailed underlying mechanism remain unanswered. In the present study, we identify the critical role of claudin-7 in Rab25-induced suppression of colon cancer invasion.
Methods
To define the role of Rab25 and claudin-7 in colon cancer cells, we performed plasmid transfection and analyzed cancer cell invasion by utilizing 2D and 3D Matrigel invasion chambers. Immunoblotting, immunofluorescence and ELISA assay were used to identify the level of protein expression and pathways implicated in Rab25-induced colon cancer cell invasion.
Results
Enforced expression of Rab25 attenuates colon cancer cell invasion. In addition, Rab25 inactivated epidermal growth factor receptor (EGFR) and increased E-cadherin expression. Unexpectedly, we observed that Rab25 induces claudin-7 expression through protein stabilization. Moreover, ectopic expression of claudin-7 reduced EGFR activity and Snail expression as well as colon cancer cell invasion. However, silencing of claudin-7 expression reversed the tumor suppressive role of Rab25, thereby increasing colon cancer cell invasiveness.
Conclusion
Collectively, the present data indicate that Rab25 inactivates EGFR and colon cancer cell invasion through upregulating claudin-7 expression.
... As described in a previous study [31], gel containing type I-P-collagen (Nitta Gelatin, Osaka, Japan) and Matrigel (Becton, Dickinson and Company) were overlaid onto the top chambers of the transwells in 24-well plates. N-/A-hPDLSCs and HNSCC cells were labeled with DiI and DiO, respectively (Thermo Fisher Scientific). ...