Expression Of Extracellular Matrix Metalloproteinase Inducer Research Articles

Identification of an active site of EMMPRIN for the augmentation of matrix metalloproteinase-1 and -3 expression in a co-culture of human uterine cervical carcinoma cells and fibroblasts

Objective Extracellular matrix metalloproteinase inducer (EMMPRIN) is highly expressed on malignant tumor cell surface and accelerates tumor invasion. We previously reported that human uterine cervical carcinoma SKG-II cells exhibit the progression of in-vitro invasiveness by utilizing the enhanced production of matrix metalloproteinase (MMP) in human uterine cervical fibroblasts (HUCF) under an in-vitro co-culture model (Sato T et al., Gynecol Oncol 2004; 92:47–56). The aim of this study was to clarify the active site of EMMPRIN in the augmentation of MMP production in the co-culture of SKG-II cells and HUCF. Methods Western and Northern blot analyses were used to examine EMMPRIN and MMP expression in a co-culture of SKG-II cells or EMMPRIN-transfected COS-7 cells and HUCF. A systematic peptide screening method using nine synthetic EMMPRIN peptides was used to identify active site(s) of EMMPRIN for MMP induction. Results SKG-II cells constitutively expressed 53-kDa EMMPRIN on the cell surface and EMMPRIN production was enhanced under the co-culture. The concomitant augmentation of proMMP-3 production was diminished by adding an EMMPRIN antibody. EMMPRIN-transfected COS-7 cells stimulated HUCF to predominantly augment proMMP-1 and -3 expressions. A systematic peptide screening method revealed that 42SLNDSATEVTGHRWLK 57 in the first loop domain of EMMPRIN participated in the augmentation of proMMP-1 production. Conclusions These results provide a novel mechanism of malignancy of uterine cervical carcinoma, in that the augmentation of EMMPRIN expression by tumor–stromal cell interaction progresses tumor invasion along with the increase of MMP expression via an active site of EMMPRIN, 42SLNDSATEVTGHRWLK 57.

Extracellular matrix metalloproteinase inducer (EMMPRIN) is present in smooth muscle cells of human aneurysmal aorta and is induced by angiotensin II in vitro

The aim of the present study was to determine whether EMMPRIN (extracellular matrix metalloproteinase inducer) is present and is up-regulated in human aneurysmal aortas, and to assess a possible association with AngII (angiotensin II)-induced aneurysm formation. The presence of EMMPRIN was assessed in 41 surgical specimens from patients with a TAA (thoracic aortic aneurysm) (Type A aortic dissection, n=12; Type B aortic dissection, n=7; and TAA without dissection, n=7) or an AAA (abdominal aortic aneurysm, n=15) by immunohistochemistry. EMMPRIN expression in aortic aneurysm tissues was compared with 12 aortas obtained during autopsy (free of any vascular diseases), and scored for both staining intensity and the percentage of vascular cells stained. EMMPRIN protein levels in cultured human aortic SMCs (smooth muscle cells) following stimulation of AngII were analysed by Western blotting. Significant EMMPRIN immunoreactivity was detected in aortic aneurysm lesions from patients with TAAs and AAAs. In the aneurysmal wall, alpha-actin-positive SMCs were the main source of EMMPRIN. The frequency of EMMPRIN overexpression was significantly higher (P=0.026) in TAAs with dissection (68.4%) than TAAs without dissection (14.3%). AngII stimulation up-regulated the expression of EMMPIRN in cultured human aortic SMCs, which was suppressed by the addition of the AT1R (AngII type 1 receptor) antagonist losartan. In conclusion, the present study is the first to report the expression of EMMPRIN in aortic aneurysmal diseases, and we speculate that EMMPRIN may be important in the pathogenesis of these diseases. Whether these abnormalities are potential therapeutic targets deserve further investigation.

Expression of extracellular matrix metalloproteinase inducer and enhancement of the production of matrix metalloproteinase-1 in tongue squamous cell carcinoma

Recent studies have found that in addition to promoting cellular invasion, overexpression of metalloproteinase -1 (MMP-1) is associated with the initial stages of cancer development. Extracellular matrix metalloproteinase inducer (EMMPRIN), a transmembrane glycoprotein, has been reported to be highly expressed in tumor cells and induce production of MMPs from peritumor fibroblasts (PTFs) adjacent to the tumor cells. The expression of EMMPRIN in tongue squamous cell carcinoma (SCC) was investigated in this study. It was found that EMMPRIN was expressed at the cell membrane throughout the entire lesion in tongue SCC. Immunofluorescence staining localized EMMPRIN to the cell membrane in a highly invasive tongue SCC cell line (Tca 8113). EMMPRIN mRNA was expressed at a high level in Tca 8113, whereas MMP-1 mRNA was expressed in PTF but harder to be detected in Tca 8113. Co-culture of Tca 8113 with PTF stimulated production of MMP-1. EMMPRIN was highly expressed in tongue SCC, and could induce local production of MMP-1. These data indicate that EMMPRIN might play an important role in tongue SCC progression and invasion.

Extracellular matrix metalloproteinase inducer is expressed in the proximal tubular epithelial cells of the human kidney

Matrix metalloproteinases (MMP) affect matrix remodelling, and extracellular matrix metalloproteinase inducer (EMMPRIN) has been reported to increase the levels of several MMP. However, the expression of EMMPRIN in the human kidney and its regulatory mechanisms are not well known. In this study, we examined EMMPRIN expression in the human kidney with the biopsied specimens, cultured proximal tubular epithelial cells (PTEC) and human mesangial cells (HMC). EMMPRIN expression was examined by immunofluorescent (IF) study, reverse transcription polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay. We also examined soluble EMMPRIN in the conditioned medium of PTEC stimulated by various agents and its effect in the activities of MMP-2 and MMP-9. Also, IF study in the several kidney diseases was performed to elucidate its role in pathological condition. EMMPRIN expression was diffusely observed in the tubular epithelial cells of most patients and healthy adults, but was never observed in glomeruli. Cultured PTEC expressed EMMPRIN, while HMC did not. Soluble EMMPRIN was also detected by enzyme-linked immunosorbent assay in the conditioned medium of PTEC. Epidermal growth factor (50 ng/mL) and phorbol 12-myristate 13-acetate (10(-7) mol/L) stimulated the secretion of soluble EMMPRIN and increased the MMP-2 activity, although these agents did not increase the level of EMMPRIN mRNA. From the IF study, EMMPRIN expression was shown to decrease in tubulointerstitial nephritis. EMMPRIN is widely distributed in the tubular epithelial cells of the adult human kidney and may regulate MMP-2 activity via its secretion from PTEC.

Increased expression of extracellular matrix metalloproteinase inducer is associated with matrix metalloproteinase‐1 and ‐2 in gingival tissues from patients with periodontitis

Extracellular matrix metalloproteinase inducer (EMMPRIN), an immunoglobulin-like cell surface glycoprotein, could promote collagenolytic balance in favor of the expression and activation of matrix metalloproteinases (MMPs). This study was to investigate the expression of EMMPRIN in gingival tissues from different periodontal conditions and to correlate it with the production of MMP-1 and MMP-2. Gingival biopsies were collected from 15 patients with untreated advanced chronic periodontitis and 15 patients with aggressive periodontitis (AgP). The control group consisted of 12 subjects diagnosed either as periodontally healthy individuals or as individuals with a gingival index of one (H/G1). The peptides and mRNA of EMMPRIN, MMP-1 and MMP-2 were detected by immunohistochemistry and semi-quantitative reverse transcriptase-polymerase chain reaction, respectively. The expression of EMMPRIN, MMP-1 and MMP-2 peptides in periodontally healthy tissues was mainly confined to the gingival epithelium. The EMMPRIN was strongly expressed in the cell membrane of the basal layer. Immunoreactivity for EMMPRIN was more intensive and more widespread in periodontitis, extended from the epithelial layers to the underlying connective tissues, and was essential in both inflammatory and fibroblast-like cells. In addition, MMP-1 and MMP-2 showed the same localized expression. The chronic periodontitis group had a significantly higher mRNA expression of EMMPRIN and MMP-2 compared with the H/G1 subjects (p < 0.05). Production of MMP-1 and MMP-2 by gingival tissues was correlated with the mRNA level of EMMPRIN (r = 0.463, p = 0.013 for MMP-1 and r = 0.404, p = 0.033 for MMP-2). The expression of EMMPRIN in human normal and diseased gingiva might contribute to periodontal physiological and pathological processes; moreover, its increased production might be associated with MMP-1 and MMP-2 expression.

Inhibiting extracellular matrix metalloproteinase inducer maybe beneficial for diminishing the atherosclerotic plaque instability

Atherosclerotic plaque rupture and local thrombosis activation in the artery cause acute serious incidents such as acute coronary syndrome and stroke. The exact mechanism of plaque rupture remains unclear but excessive degradation of the extracellular matrix scaffold by matrix-degrading metalloproteinases (MMPs) has been implicated as one of the major molecular mechanisms in this process. Convincing evidence is available to prove that extracellular matrix metalloproteinase inducer (EMMPRIN) induces MMP expression and is involved in the inflammatory responses in the artery wall. The inflammation and MMPs have been shown to play a critical role for atherosclerotic lesion development and progression. More recent data showed that increased EMMPRIN expression was associated with vulnerable atherosclerotic lesions. Therefore, we speculate that EMMPRIN may be pivotal for atherosclerotic plaque instability, and hence inhibition of EMMPRIN expression could be a promising approach for the prevention or treatment of atheroma instability.

Expression of Fibrinolytic- and Matrix Degradation and Remodeling-Related Genes in Injured Peritoneum that Healed with Adhesions and without Adhesions in a Mouse Model

Study Objective: To compare the mRNA levels of tissue-type plasminogen activator (tPA), plasminogen activator inhibitor 1 (PAI-1), extracellular matrix metalloproteinase inducer (EMMPRIN) and tissue inhibitor of metalloproteinases-1 (TIMP-1) among injured peritoneum that healed with adhesions, injured peritoneum that healed without adhesions, and normal non-injured peritoneum. Design: Prospective experimental study. Setting: Academic facility. Patients: Adult female Balb/c and C57BL6 mice. Intervention: On day 0, a laparotomy was performed to develop peritoneal adhesions in C57BJ6 (n 5 10) and Balb/c mice (n 5 10). On day 7, a second laparotomy was performed to assess adhesion formation. Measurements and Main Results: Expression levels of tPA, PAI-1, EMMPRIN and TIMP-1 mRNA in injured peritoneum that healed with adhesions, injured peritoneum that healed without adhesions, and normal non-injured peritoneum.Expression levels of tPA and PAI-1 were significantly higher in injured peritoneum that healed with adhesions than in either peritoneum without adhesions or non-injured normal peritoneum, in both strains. TIMP-1 expression was significantly higher in injured peritoneum regardless of adhesions than in non-injured normal peritoneum in both strains. There was no significant difference in EMMPRIN expression among any groups in either strain. Conclusion: The present study demonstrated that tPA and PAI-1 mRNA were differentially expressed in injured peritoneum that healed with adhesions and without adhesions in both Balb/c and C57BL6 mice.

Cytoplasmic and Nuclear Maspin Expression in Lung Carcinomas

Maspin, a serine protease inhibitor related to the serpin family, can inhibit invasion and metastasis of malignancies although direct evidence of the clinicopathologic significance of cytoplasmic relative to nuclear expression is limited. Here, maspin expression was examined on tissue microarrays containing lung carcinoma (n=155) and adjacent noncancerous tissue (n=20) and also 4 lung carcinoma cell lines (LC-1/Sq, LC-IF, PC-14, and AoI) by immunohistochemistry. Maspin expression was compared with clinicopathologic parameters of the tumors. Maspin expression showed positive nuclear staining in basal cells, LC-IF, and PC-14 cell lines, and also cytoplasmic immunoreactivity in secretory and ciliated cells, LC-1/Sq cell line. Cytoplasmic staining was the lowest in adenocarcinoma (AD) and the highest in squamous cell carcinoma as compared with other types of lung carcinoma (P<0.05), and positively correlated with expression of p53 and caspase-3 (P<0.05). The cytoplasmic one showed stronger immunoreactivity in male carcinoma patients than female ones (P<0.05). The nuclear maspin expression gradually increased through squamous cell carcinoma, AD, large cell carcinoma to small cell carcinoma (P<0.05) and was also positively associated with the levels of vascular epithelial growth factor and extracellular matrix metalloproteinase inducer expression (P<0.05). Kaplan-Meier analysis indicated that the cytoplasmic or nuclear maspin expression was not a good prognostic marker for lung carcinomas overall (P>0.05), but the cytoplasmic pattern pointed to good survival for AD cases (P<0.05). It was concluded that the cytoplasmic and nuclear expression patterns of maspin are involved in the cellular differentiation of normal lung tissue and the histogenesis of different lung carcinomas. The cytoplasmic maspin may play an important role in lung carcinomas by regulating apoptosis and thus is a favorable prognostic marker for AD patients, whereas the nuclear location may be linked to promotion of angiogenesis.

Clinical implications and prognostic value of EMMPRIN/CD147 and MMP2 expression in pediatric gliomas

Extracellular matrix metalloproteinase inducer (EMMPRIN), a member of the immunoglobulin superfamily, is present on the surface of tumor cells where it stimulates adjacent fibroblasts to produce matrix metalloproteinases (MMPs). We have analyzed the clinicopathological characteristics of EMMPRIN and MMP2 expression in normal brain tissue and pediatric gliomas and evaluated their prognostic value in diagnosing the latter. Immunochemistry analysis revealed EMMPRIN and MMP2 expression in cryo-sections of pediatric gliomas (45 samples) and normal brain tissue (20 samples). Both EMMPRIN and MMP2 were expressed in normal brain and glioma tissues with different levels of malignancy. The intensively positive expression rates of EMMPRIN (22/27) and MMP2 (21/27) in anaplastic astrocytoma and glioblastoma tissues were significantly higher than those in normal brain and low-grade astrocytoma tissues (2/28 and (1/2)8, respectively). Spearman analysis indicated that the expression level of EMMPRIN was significantly positively correlated with that of MMP2 (r = 0.86, p < 0.01). The positive expression of EMMPRIN and MMP2 was associated with higher grade gliomas. Patients with EMMPRIN+/MMP2+ expression had the lowest survival rate (p < 0.01). Based on these results, we conclude that EMMPRIN and MMP2 are expressed differently in normal brain and pediatric gliomas. The detection of their co-expression may facilitate the prediction of pediatric gliomas prognosis.

Pulmonary large cell carcinoma displays high expression of EMMPRIN and VEGF

To investigate the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) and vascular endothelial growth factor (VEGF) in lung carcinomas, and to clarify their roles in carcinoma progression. Expression of EMMPRIN and VEGF was examined with tissue microarrays (TMAs) of lung carcinomas (n = 181), and their suppression in adjacent normal lung samples (n = 40) were determined by immunohistochemistry. The results were compared with clinicopathological findings for the same tumors. Both EMMPRIN and VEGF were occasionally expressed in pseudostratified columnar epithelium and frequently in lung carcinomas. Histologically, EMMPRIN and VEGF displayed higher levels in large (LCC) cell carcinomas than adenocarcinoma (AD), squamous (SQ) and small cell carcinomas (SCC) (P < 0.05). EMMPRIN was more highly expressed in SQ as compared with AD (P < 0.05), while the converse was true for VEGF (P < 0.05). Binding was generally more intense for EMMPRIN in samples from male compared to female patients (P < 0.05), whereas the latter tended to exhibit more VEGF expression (P < 0.05). Positive associations of VEGF expression with the TNM stage and amounts of EMMPRIN were noted in the lung carcinomas (P < 0.05). EMMPRIN and VEGF possibly contribute to physiological repair of normal lung and histogenesis of lung carcinoma. Both proteins might be involved in the molecular basis for diff erences in the incidence of lung carcinoma between men and women.

Extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinases (MMPs) as regulators of tumor–host interaction in a spontaneous metastasis model in rats

EMMPRIN has a role in invasion and metastasis through the induction of MMPs and the consequent modulation of cell-substrate and cell-cell adhesion processes. The present study evaluates the expression of EMMPRIN protein and MMP-2/9 activity in tumor and parenchymal cells in a spontaneous metastasis model in rats. Moreover, we explore the regulation of EMMPRIN and MMP-9 by tumor-epithelial cell interactions in vitro. By zymography, we observed an increased proMMP-9 expression in both metastasized liver and spleen samples from tumor bearing rats. Immunohistochemical studies showed EMMPRIN-positive tumor cells in tumor biopsies as well as in spleen and liver samples from tumor bearing rats. Interestingly, a significant increase in EMMPRIN expression in hepatic cells was also detected. The regulation of EMMPRIN expression in tumor and liver cells in response to tumor-host interaction was investigated in vitro through a tumor cell line culture on extracellular matrix (ECM) molecules or in co-culture with normal rat liver cells (BRL3A cells). No significant changes in EMMPRIN expression were detected in tumor cells cultured on ECM molecules. On the other hand, EMMPRIN protein and MMP-9 mRNA expression were induced in BRL3A cells. The increase in EMMPRIN expression in BRL3A cells was inhibited by an anti-EMMPRIN antibody. These results reinforce the main role of EMMPRIN mediating tumor-host interactions that may evolve new opportunities for therapeutic interventions.

Matrix metalloproteinases 2 and 9 as the factors of head and neck tumor metastases

The levels of metalloproteinases (MMP-2,-9), their tissue inhibitors (TIMP-1,-2) and extracellular matrix metalloproteinase inducer (EMMPRIN) were studied in tumor tissue and blood serum from patients with head and neck squamous sell carcinoma. Immunohistochemical investigation showed much higher expression of MMP-9 and TIMP-1 in tumor tissue than MMP-2 and TIMP-2. Different expression of the studied parameters (except TIMP-1) in cancer cells and stroma was observed. Enhanced level of MMP-2,-9 and TIMP-2 expression was revealed mainly in fibrocytes, leukocytes and stromal extracellular matrix. Expression of EMMPRIN in tumor cells was higher than in stromal cells. It is possible that carcinoma cells express EMMPRIN to increase MMP production by surrounding cells. Significant decrease of inhibitor expression was observed in cells of carcinomas with N1 grade of metastasis than in tumors without metastasis. Also the decrease in TIMP-1 level was determined in blood serum of patients with metastasis to regional lymph nodes compared with serum of patients without metastasis. Thus, MMP-9 and TIMP-1 play an important role in the development of head and neck squamous sell carcinoma and TIMP-1 level in blood serum and cancer tissues is linked to first grade of regional lymph node metastasis.

Resveratrol inhibits EMMPRIN expression via P38 and ERK1/2 pathways in PMA-induced THP-1 cells

The extracellular matrix metalloproteinase inducer (EMMPRIN) is significant in the regulation of matrix metalloproteinase (MMP) synthesis in atherosclerosis-related cells, and is possibly involved in the progression of atherosclerotic plaque. EMMPRIN expression is also up-regulated in PMA-induced THP-1 cells and is inhibited by resveratrol. However, it remains unclear how resveratrol inhibits EMMPRIN expression. We thus investigated the role of the MAPK signaling pathway in resveratrol inhibiting the up-regulation of EMMPRIN in PMA-induced THP-1 cells. We found that the ERK1/2 and p38 pathways, but not the JNK, are activated during the up-regulation of EMMPRIN expression. We also observed that while resveratrol suppresses the up-regulation of EMMPRIN, it also suppresses both the ERK1/2 and p38 pathways in a dose-dependent manner. Taken together, we established that it is through both the ERK1/2 and p38 MAPK pathways that resveratrol inhibits the expression of EMMPRIN in PMA-induced THP-1 cells.

Expression of extracellular matrix metalloproteinase inducer in odontogenic cysts

Extracellular matrix metalloproteinase inducer (EMMPRIN) is known to induce matrix metalloproteinase (MMP) production. The expression of EMMPRIN in odontogenic cysts has not been previously studied. This study was done to determine the presence and the variability of EMMPRIN expression in various types of odontogenic cysts. An immunohistochemical study using a polyclonal anti-EMMPRIN antibody was done using 48 odontogenic cyst cases: 13 odontogenic keratocysts (OKCs), 18 dentigerous cysts (DCs), and 17 periapical cysts (PAs). Twelve cases of normal dental follicles (DFs) were also included in this study for comparison. EMMPRIN immunoreactivity was detected in all of the cysts and DFs studied. In odontogenic cysts, EMMPRIN immunoreactivity was generally higher in basal cells than in suprabasal cells. The overall EMMPRIN expression in the epithelial lining of the 3 different types of odontogenic cyst was significantly higher than in the DFs. Overall EMMPRIN expression was also found to be significantly higher in the epithelial lining of OKCs than in the other types of cysts. This study confirmed that EMMPRIN is present in odontogenic cysts and DFs. The higher EMMPRIN expression in OKCs suggests that it may be involved in the aggressive behavior of this type of cyst.

Increased expression of matrix metalloproteinase-2 (MMP-2) predicts tumour recurrence and unfavourable outcome in non-small cell lung cancer.

The purpose of this study was to analyse the expression of matrix metalloproteinase-2 (MMP-2) and its extracellular matrix metalloproteinase inducer (EMMPRIN) in non-small cell lung cancer (NSCLC), and to evaluate their significance to predict tumour behaviour. The study consists of 212 patients treated by the resection of the tumour. Tumour samples were stained immunohistochemically, and the expression of MMP-2 and EMMPRIN was evaluated both in tumour cells and in peritumoural stromal tissue. The results were compared with clinicopathological factors and survival of the patients. High expression of MMP-2 in tumour cells was found in 83 out of 191 cases (44%). Adenocarcinomas showed more often high expression of MMP-2 as compared with squamous cell or large cell carcinomas (p=0.001). High cancer cell associated MMP-2 expression was associated with increased tumour recurrence (p=0.001). Tumour stroma showed positive staining in 162 (98%) cases and was considered highly stained in 120 (72%) cases. The high stromal MMP-2 expression was noticed more often among large cell carcinomas as compared with other histological types (p=0.007). High cancer cell associated EMMPRIN expression was found in 115 (61%) cases and was associated only with high MMP-2 expression in tumour cells (p=0.006). In overall survival (OS) and disease free survival (DFS) analyses, type of tumour (p=0.001 and p=0.0004), advanced stage (p=0.001 and p=0.013) and high MMP-2 expression in tumour cells (p=0.018 and p=0.001) were associated with poor survival. Also, high stromal MMP-2 expression was related to poor outcome in both OS and DFS analyses (p=0.010 and 0.045, respectively). In multivariate analysis, stromal MMP-2 expression retained its prognostic value to predict OS and DFS (p=0.028 and p=0.039, respectively), together with tumour type and stage (p=0.017, p=0.001 and p=0.021, p=0.008, respectively). The present study shows the significant prognostic value of MMP-2 in NSCLC suggesting that the use of MMP-2 is valuable in determining the patients with more aggressive disease.

Mixed-type gastric carcinomas exhibit more aggressive features and indicate the histogenesis of carcinomas

To investigate the pathobiological behaviors of gastric mixed-type (MT) carcinomas and gastric carcinogenesis, the clinicopathological characteristics of MT carcinomas were analyzed and compared with intestinal-type (IT) and diffuse-type (DT) carcinomas. The expression of Ki-67, caspase-3, p53, fragile histine triad (FHIT), maspin, extracellular matrix metalloproteinase inducer (EMMPRIN), vascular growth factor (VEGF), MUC-2, 4, 5AC and 6, CD44, E-cadherin, β-catenin, and phosphorylated glycogen synthase kinase 3β-ser9 (P-GSK3β-ser9) was examined on tissue microarrays using immunohistochemistry. It was found that MT carcinomas exhibited large size, deep invasion, frequent local invasion, and lymph node metastasis in comparison with IT and DT carcinomas (p < 0.05). All the markers except MUC-5AC showed higher expression in IT than DT carcinomas (p < 0.05). The expression of maspin, EMMPRIN, VEGF, MUC-4, and membrane E-cadherin was stronger in MT intestinal than diffuse component (p < 0.05). Immunoreactivities to Ki-67, EMMPRIN, and VEGF were weaker in IT carcinoma than in the MT intestinal portion (p < 0.05), while the opposite was true for CD44, MUC-2, and MUC-6 (p < 0.05). The MT diffuse component displayed a higher expression of FHIT, VEGF, and P-GSK3β-ser9 than DT carcinoma (p < 0.05). The accumulative survival rate of the IT carcinoma patients was higher than the other types (p < 0.05). The invasive depth, venous invasion, lymph node, peritoneal or liver metastasis, and Lauren's classification were independent prognostic factors for gastric carcinomas (p < 0.05). These findings suggested that MT carcinomas were also indicated to be more aggressive than IT and DT carcinomas. Significant differences were observed in the proliferation, apoptosis, angiogenesis, mucin secretion, and cell adhesion between IT and DT carcinomas, whereas only a few of these characteristics showed differences between the MT intestinal and diffuse parts, thus suggesting that both the MT components might originate from the stem cells with similar genetic traits, but follow different histogenic pathways.

Role of basement membrane in EMMPRIN/CD147 induction in rat tracheal epithelial cells

Extracellular matrix metalloproteinase inducer (EMMPRIN) is a glycosylated transmembrane protein known to induce matrix metalloproteinases (MMPs). Although the expression of EMMPRIN is physiologically limited to fetal lung epithelium, the transcriptional regulation of this protein remains to be elucidated. We hypothesized that the interaction of epithelial cells with the basement membrane regulates EMMPRIN expression. The basement membrane has highly integrated architecture composed of specific extracellular matrix, such as laminins and type IV collagen, and exhibits multiple functions. We previously developed a structured basement membrane mimic, a synthesized basement membrane (sBM) substratum, in which laminin-111, a unique component of embryonic lungs, is incorporated. In the present study we quantified expression of EMMPRIN mRNA of rat tracheal epithelial cells cultured on sBM, laminin-111, type IV collagen, or laminin-332. EMMPRIN was upregulated on sBM and laminin-111, although this was not accompanied by MMP-9 induction. In contrast, type IV collagen and laminin-332 did not induce EMMPRIN. These findings suggest potential roles for basement membrane in the transcriptional regulation of tracheal epithelial EMMPRIN.

Selective progesterone receptor modulator asoprisnil down-regulates collagen synthesis in cultured human uterine leiomyoma cells through up-regulating extracellular matrix metalloproteinase inducer

A recent clinical trial demonstrated that selective progesterone receptor modulator asoprisnil is effective in reducing uterine leiomyoma volume. We investigated the effects of asoprisnil in vitro on the expression of the extracellular matrix (ECM)-remodeling enzymes and collagens in cultured leiomyoma and matching normal myometrial cells. The expression of extracellular matrix metalloproteinase inducer (EMMPRIN), matrix metalloproteinases (MMPs), tissue inhibitors of MMP (TIMPs) and collagens were assessed by western blot analysis. Untreated cultured leiomyoma cells had significantly lower EMMPRIN (P < 0.05), MMP-1 (P < 0.05) and membrane type 1-MMP (MT1-MMP) (P < 0.01) protein contents, but significantly higher TIMP-1 (P < 0.05), TIMP-2 (P < 0.01), type I (P < 0.05) and type III (P < 0.01) collagen protein contents compared with untreated cultured myometrial cells. Treatment with asoprisnil at concentrations > or =10(-7) M for 48 h significantly (P < 0.05) increased EMMPRIN, MMP-1 and MT1-MMP protein contents, and decreased TIMP-1 (P < 0.05), TIMP-2 (P < 0.01), type I (P < 0.01) and type III (P < 0.05 at 10(-7) M; P < 0.01 at 10(-6) M) collagen protein contents in cultured leiomyoma cells compared with control cultures. However, asoprisnil treatment did not affect the protein contents of ECM-remodeling enzymes and collagens in cultured myometrial cells. These results suggest that asoprisnil may reduce collagen deposit in the ECM of cultured leiomyoma cells through decreasing collagen synthesis and enhancing the expression of EMMPRIN, MMPs and TIMPs without comparable effects on cultured myometrial cells.

Progesterone receptor modulator CDB-2914 induces extracellular matrix metalloproteinase inducer in cultured human uterine leiomyoma cells

Effects of progesterone receptor modulator CDB-2914 on the expression of the extracellular matrix (ECM) components were examined in cultured human uterine leiomyoma and myometrial cells. ECM metalloproteinase inducer (EMMPRIN), matrix metalloproteinases (MMPs), tissue inhibitors of MMP (TIMPs) and collagen levels were assessed by Western blot analysis, MMP activity assay and real-time RT-PCR. RNA interference (RNAi) of EMMPRIN was performed using small interfering mRNA. In cultured leiomyoma cells, CDB-2914 treatment at concentrations greater than or equal to 10(-8) M significantly increased EMMPRIN, MMP-1 and MMP-8 protein contents and MMP-1, MMP-2, MMP-3 and MMP-9 mRNA levels, and activity of MMP-1, MMP-2, MMP-3 and MMP-9 in the medium. TIMP-1 and TIMP-2 were significantly decreased at mRNA and protein levels by CDB-2914 treatment at concentrations > or =10(-7) M in these cells. CDB-2914 treatment decreased types I and III collagen protein contents. However, CDB-2914 treatment did not affect the ECM component expression in cultured myometrial cells. RNAi of EMMPRIN abrogated CDB-2914-mediated both induction of MMPs and reduction of TIMPs and collagens in cultured leiomyoma cells. These results suggest that CDB-2914 modulates the expression of EMMPRIN, MMPs, TIMPs and collagens in cultured leiomyoma cells without comparable effects on myometrial cells.

Pancreatic stellate cells—role in pancreas cancer

Adenocarcinomas of the pancreas are characterized by a rapid progression, an early metastasis, a limited response to chemo- and radiotherapy, and an intense fibrotic reaction known as tumor desmoplasia. Carcinoma cells are surrounded by a dense stroma consisting of myofibroblast-like cells, collagens, and fibronectin. This review describes the interaction of activated pancreatic stellate cells (myofibroblast-like cells) with tumor cells in pancreas adenocarcinomas. Our data were obtained in cell culture experiments and in in vivo investigations. Carcinoma cells produce soluble mediators and stimulate motility, proliferation, matrix-, and MMP synthesis of stellate cells. Vice versa-activated stellate cells release mitogens, stimulating proliferation of cancer cells. Cancer cell proliferation and resistance to apoptosis might further be induced by the microenvironment (extracellular matrix), which is primarily provided by stellate cells. A very important aspect in the interaction of stellate cells with cancer cells is the expression of EMMPRIN (extracellular matrix metalloproteinase inducer) by cancer cells, the shedding of the extracellular part of EMMPRIN by matrix metalloproteinases (MMPs), and the induction of MMPs in stellate cells by soluble EMMPRIN. In particular, the stellate cells in close proximity to tumor cells therefore express MMPs and degrade connective tissue. Through complex interactions between stellate cells and carcinoma cells, tumor progression and cancer cell invasion are accelerated. As we gain better understanding of these mechanisms, adequate therapies to reduce tumor cell invasion and cancer progression might be developed.