The expression of six biomarkers in the four most common ovarian cancers: correlation with clinicopathological parameters

This study aimed to evaluate the relationship of fascin‐1, matrix metalloproteinase (MMP)‐2, MMP‐9, cortactin, survivin, and epidermal growth factor receptor (EGFR) expression with clinicopathological parameters for the four most common ovarian surface epithelial carcinomas. Six biomarkers were investigated immunohistochemically using tissue microarrays of 185 specimens including 79 serous cystadenocarcinomas, 47 mucinous cystadenocarcinomas, 45 endometrioid adenocarcinomas, and 14 clear cell carcinomas. The four most common ovarian carcinomas showed significant expression of fascin‐1, cortactin, survivin, and EGFR, but not of MMP‐2 and MMP‐9. In addition, higher immunostaining scores for fascin‐1 in mucinous cystadenocarcinomas correlated with T stage, N stage, American Joint Committee on Cancer AJCC clinical stage, and a poorer survival rate; for cortactin in serous cystadenocarcinomas correlated with T stage; for cortactin in clear cell carcinomas correlated with T and clinical AJCC stages; and for survivin in clear cell carcinomas correlated with T stage and AJCC clinical stage. In addition, higher immunostaining scores for fascin‐1, cortactin, and survivin correlated with poorer tumor differentiation in serous, mucinous, and endometrioid adenocarcinomas. Thus, the expression of fascin‐1, cortactin, and survivin may be helpful in evaluating the aggressiveness of ovarian mucinous, serous, and clear cell adenocarcinoma. Additionally, the expression of fascin‐1 may be an independent prognostic risk factor in mucinous cystadenocarcinoma.     

Study of molecular prognostic factors Bcl-2 and EGFR in rectal mucinous carcinomas

Colorectal cancer is an important public health problem worldwide, being the fourth most common cancer in men and the third most common in women. Colorectal cancer incidence is higher in developing countries due to the prevalence of obesity associated with reduced physical activity. Rectal mucinous carcinomas, especially the "signet ring cell" type, have a worse prognosis compared with other varieties of colorectal carcinomas, being diagnosed in more advanced stage and more prone to lymph node and peritoneal metastases. Our study comprised 37 cases with rectal adenocarcinoma with mucinous component operated in the Surgical Clinics of the Emergency County Hospital of Craiova, between 2006 and 2010. The aim of this study was to evaluate some molecular prognostic factors for rectal mucinous carcinomas namely B-cell lymphoma 2 (Bcl-2) and epidermal growth factor receptor (EGFR), and their correlations with the main morpho-clinical parameters of these patients. Thus, we immunohistochemically assessed the degree of apoptosis of tumor cells in mucinous rectal carcinomas using the Bcl-2 marker, and tumor aggressiveness using the EGFR responsiveness. In our study, the percentage of Bcl-2+ cases was 43.24%, with no significant statistical correlation between the Bcl-2 expression and histopathological subtype of mucinous adenocarcinoma. The evaluation of tumor aggressiveness in terms of EGFR responsiveness showed a reduced expression in carcinomas correlated with the increase in quantity of the mucinous component. In addition, EGFR reactivity was increased in the tumor invasion front.     

Vitexin alleviates interleukin‐1β‐induced inflammatory responses in chondrocytes from osteoarthritis patients: Involvement of HIF‐1α pathway

It has been reported that vitexin has anti‐inflammatory effects in osteoarthritis (OA) rats. However, the effects of vitexin on interleukins‐1β (IL‐1β)‐stimulated OA patient‐derived chondrocytes have not been reported. The purpose of this study was to investigate the anti‐inflammatory effects of vitexin on IL‐1β‐stimulated human osteoarthritis chondrocytes and to reveal the involvement of hypoxia‐inducible factor 1α (HIF‐1α) pathway. Enzyme‐linked immunosorbent assay, quantitative real‐time PCR and Western blotting assays were employed. ELISA results demonstrated that the proinflammatory cytokine levels of interleukins‐6 (IL‐6) and tumour necrosis factor α (TNF‐α) in the serum and synovial fluid and HIF‐1α level in the synovial fluid were significantly elevated in OA patients compared to normal healthy subjects. Moreover, the Western blotting results indicated that the protein expression of HIF‐1α was significantly higher in the cartilage tissues of OA patients. OA patient‐derived chondrocytes were stimulated by IL‐1β and treated with different concentration of vitexin for 24 hours. Vitexin showed no cytotoxicity and increased the survival of chondrocytes under IL‐1β stimulation. Vitexin suppressed IL‐1β‐induced production of NO and prostaglandin E2 (PGE₂) in chondrocytes culture. The treatment of vitexin significantly inhibited IL‐1β‐induced expressions of proinflammatory cytokine levels of IL‐6, TNF‐α, matrix metalloproteinase (MMP)‐1, MMP‐3 and MMP‐13. Furthermore, Western blotting results demonstrated that HIF‐1α is involved in vitexin's protective effects on IL‐1β‐stimulated injuries in OA patient‐derived chondrocytes. Our study demonstrates that vitexin alleviates IL‐1β‐induced inflammatory responses in chondrocytes from osteoarthritis patients, which may be attributed partly to the inhibition of HIF‐1α pathway.     

Colorectal signet ring cell carcinoma: Influence of EGFR,E-cadherin and MMP-13 expression on clinicopathological features and prognosis

Signet ring cell carcinoma (SRCC) is unique rare subtype of mucin-producing colorectal adenocarcinoma characterized by presence of signet ring cells, in > 50% of the tumor tissue. This study aims to investigate expression of EGFR, E-cadherin and MMP-13 expression on clinicopathological features of signet ring cell type and its prognostic effect using manual tissue microarray technique. In this work, we studied tumor tissue specimens from 150 patients with colorectal cancer cases among which 19 cases of SRCC. High density manual tissue microarrays were constructed using modified mechanical pencil tips technique and immunohistochemistry for EGFR, E-cadherin and MMP-13 expression was done. We found that SRCC was significantly associated with younger age and more frequency of LN metastasis than all other groups. SRCC was also significantly associated with annular gross picture, more depth of invasion, advanced stage, more lymphovascular emboli, more perineural invasion and less arousal from an overlying adenoma. In conclusion, colorectal SRCC has distinctive clinicopathological and histological features with different unique mechanisms of carcinogenesis and more aggressive biologic behavior than other colorectal carcinoma subtypes. Negative/low expressions of EGFR and E-cadherin and MMP-13 were found in SRCC with no effect on the prognosis.     

Involvement of α‐ and β‐catenins and E‐cadherin in the development of mammary phyllodes tumours

Tsang J Y S, Mendoza P, Lam C C F, Yu A M C, Putti T C, Karim R Z, Scolyer R A, Lee C S, Tan P H & Tse G M
(2012) Histopathology  61, 667–674 Involvement of α‐ and β‐catenins and E‐cadherin in the development of mammary phyllodes tumours Aims: Phyllodes tumours (PT) are rare but clinically important fibroepithelial tumours of the breast. β‐Catenin, a key component in Wnt signalling, has been shown to be important in the development of PT. It also functions as a component of the cadherin complex, which may therefore be implicated in PT pathogenesis. By assessing stromal α‐catenin, β‐catenin and E‐cadherin expression in 158 PT cases using immunohistochemistry and examining associations with clinicopathological features, we aimed to determine the role of these proteins in PT pathogenesis. Methods and results: Cytoplasmic β‐catenin correlated with α‐catenin expression. A significantly higher expression of both markers was observed in borderline than in benign PT (P = 0.003 and <0.001, respectively), but a lower level was found in malignant PT. Cytoplasmic E‐cadherin expression was significantly higher in borderline and malignant than in benign PT (P = 0.001 and 0.012, respectively), but was not correlated with other markers. Both E‐cadherin and α‐catenin showed stronger correlations with histological parameters than β‐catenin. α‐Catenin showed a significant correlation with recurrence (P = 0.005 and 0.016, respectively). Conclusions: α‐ and β‐catenins may be important in the early stages of PT development, while E‐cadherin may be required for malignant development. The correlation of α‐catenin expression with tumour recurrence may be relevant in predicting PT behaviour.     

Systematic assessment of protein phenotypes characterizing high‐grade tumour budding in mismatch repair‐proficient colorectal cancer

Karamitopoulou E, Lugli A, Panayiotides I, Karakitsos P, Peros G, Rallis G, Patsouris E S, Terracciano L & Zlobec I (2010) Histopathology 57 , 233–243
Systematic assessment of protein phenotypes characterizing high‐grade tumour budding in mismatch repair‐proficient colorectal cancer Aims: A tumour bud is defined as a single tumour cell or tumour cell cluster of up to five cells at the invasive tumour front. Significant differences in survival have been detected in colorectal cancer patients with low‐ compared to high‐grade budding. The aim of this study was to identify potential multi‐marker phenotypes characterizing low‐ and high‐grade budding in mismatch repair (MMR)‐proficient colorectal cancer. Methods and results: Established and promising prognostic proteins such as epidermal growth factor receptor (EGFR), pERK, RHAMM, RKIP, β‐catenin, E‐cadherin, pAKT, p16, p21, Ki67, Bcl‐2, vascular endothelial growth factor (VEGF), apoptotic protease activating factor‐1 (APAF‐1), MUC1, EphB2, matrix metalloproteinase 7, pSMAD2, CDX2, laminin5γ2 and MST1 were analysed on 208 MMR‐proficient colorectal cancers with complete clinicopathological data. The most accurate markers for predicting high‐grade budding (more than six tumour buds) were EphB2 (P < 0.001), Bcl‐2 (P < 0.001), RKIP (P < 0.001), E‐cadherin (P = 0.004), laminin5γ2 (P = 0.004) and APAF‐1 (P = 0.005). On multivariable analysis, only loss of Bcl‐2 (P < 0.001) and EphB2 (P < 0.001) were independent predictors of high‐grade budding. Bcl‐2?/EphB2? tumours were more frequently poorly differentiated (P < 0.001), of advanced pT stage (P = 0.002), lymph node positive (P = 0.023), presented vascular (P = 0.053) and lymphatic invasion (P = 0.005) and had a negative impact on patient survival (P = 0.012). Conclusions: The multi‐marker phenotype EphB2?/Bcl‐2? is an independent predictor of high‐grade budding and implies increased aggressive behaviour in MMR‐proficient colorectal cancer.     

Co‐expression of TLR‐9 and MMP‐13 is associated with the degree of tumour differentiation in prostate cancer

In vitro experiments demonstrated that stimulation of Toll‐like receptor 9 (TLR‐9) by synthetic TLR‐9 ligands induces the invasion of TLR‐9‐expressing prostate cancer cells through matrix metalloproteinase 13 (MMP‐13). However, the clinical value of TLR‐9 and MMP‐13 co‐expression in the pathophysiology of the prostate is unknown. In the study, we evaluated the expression levels and clinical significance of the TLR‐9 and MMP‐13 in a series of prostate tissues. One hundred and eighty prostate tissues including prostate cancer (PCa) (n = 137), high‐grade prostatic intraepithelial neoplasia (HPIN) (n = 18) and benign prostatic hyperplasia (BPH) (n = 25) were immunostained for the TLR‐9 and MMP‐13 markers. Subsequently, the correlation between the TLR‐9 and MMP‐13 staining scores and clinicopathological parameters was obtained. Higher expressions of TLR‐9 and MMP‐13 were found in PCa and high‐grade prostatic intraepithelial neoplasia compared to benign prostatic hyperplasia tissues. Among PCa samples, a positive relationship was revealed between the MMP‐13 expression and Gleason score (P < 0.001). There was a significant correlation between TLR‐9 expression and regional lymph node involvement (P = 0.04). The expression patterns of TLR‐9 and MMP‐13 markers demonstrated a reciprocal significant correlation between the two markers in the same series of prostate samples (P < 0.001). Furthermore, the Gleason score of TLR‐9^high/MMP‐13^high and TLR‐9^low/MMP‐13^low phenotypes showed a significant difference (P = 0.002). Higher expressions of TLR‐9 and MMP‐13 can confer aggressive behaviour to PCa. Therefore, these markers may be used as a valuable target for tailored therapy of PCa.     

Clinicopathological characteristics and prognostic factors of advanced colorectal mucinous adenocarcinoma

Yamaguchi T, Taniguchi H, Fujita S, Sekine S, Yamamoto S, Akasu T, Kushima R, Tani T, Moriya Y & Shimoda T
(2012) Histopathology  61, 162–169 Clinicopathological characteristics and prognostic factors of advanced colorectal mucinous adenocarcinoma Aims: Mucinous adenocarcinoma (MUC) is a histological variant of colorectal adenocarcinoma. The aim of the present study was to characterize clinicopathological features and identify prognostic factors of MUCs. Methods and results: A total of 181 patients with MUC who underwent surgery between 1975 and 2003 were reviewed. The clinicopathological features of these patients were compared with those of 4125 non‐MUC patients. Univariate and multivariate analyses were conducted to identify significant prognostic factors in 102 patients with pT3 or pT4 tumour who underwent curative surgery. Patients with MUCs tended to present with more advanced clinical stages. The overall 5‐year survival rate of MUC patients was lower than that of non‐MUC patients; however, no prognostic difference was found when patients with the same clinical stages were compared. Multivariate analysis revealed male sex, bowel obstruction and infiltrating growth type as independent prognostic factors. Five‐year cancer‐specific survival rates for MUC patients with ≤1, 2 and 3 risk factors identified by multivariate analysis were 95.5%, 52.1% and 0.0%, respectively (P < 0.001). Conclusions: Mucinous adenocarcinoma represents a distinct clinicopathological entity. Sex, bowel obstruction and growth patterns might be useful prognostic factors to identify patients with a high risk of recurrence after curative resection of advanced MUCs.     

Prognostic significance of single isolated cells with decreased E‐cadherin expression in pseudomyxoma peritonei

Pseudomyxoma peritonei (PMP) cases can be classified into the prognosis‐related subtypes of disseminated peritoneal adenomucinosis (DPAM) and peritoneal mucinous carcinomatosis (PMCA). To investigate the mechanisms of mucinous invasion and the differing prognoses of these two subtypes, we examined the expression levels of proteins involved in cellular adhesion and invasion, including E‐cadherin, vimentin, β‐catenin, and S100A4, in single isolated tumor cells (SICs) and cohesive cellular strips within mucin pools isolated from DPAM (n = 31) and PMCA (n = 21) patients. In both PMCA and DPAM cases, SICs showed a complete loss of E‐cadherin expression, whereas cells in cohesive cellular clusters retained E‐cadherin expression. The frequency of high numbers of SICs (>8) in PMCA cases was significantly greater than that in DPAM cases (86% and 26%, respectively) and was correlated with poor progression‐free survival (P = 0.019) in a univariate analysis. In both PMP subtypes, strong vimentin expression was identified in most of the SICs but not the cohesive cellular strips. The relatively slow progression of DPAM may be attributable to the smaller number of SICs that lack E‐cadherin expression and have increased vimentin expression, whereas the rapid progression of PMCA may be due to larger numbers of these SICs.     

错配修复蛋白表达及RAS基因突变与结直肠癌临床病理特征的关系

目的探讨结直肠癌组织中错配修复(mismatch repair,MMR)蛋白表达及RAS基因突变状态与临床病理特征的相关性。方法采用免疫组化法检测186例结直肠癌组织中4种MMR蛋白(MSH2、MSH6、MLH1、PMS2)的表达,并采用ARMS-PCR法检测RAS基因突变状态。结果186例结直肠癌组织中MMR蛋白表达缺失率为6.99%(13/186),其中MSH2和MSH6共同缺失2例,MLH1和PMS2共同缺失9例,MSH6单独缺失1例,PMS2单独缺失1例。RAS突变者97例(52.15%),其中KRAS突变88例(47.31%),NRAS突变9例(4.84%)。MMR蛋白表达缺失多发生于50岁以下、肿瘤部位多为右半结肠、黏液腺癌、肿瘤低分化、肿瘤分期Ⅰ+Ⅱ期的患者(P<0.05)。RAS突变状态与结直肠癌临床病理特征无相关性(P>0.05)。MMR蛋白表达缺失病例中,RAS突变状态与肿瘤是否伴黏液腺癌成分有显著相关性(P=0.015),伴RAS突变病例不伴有黏液腺癌,而RAS野生型多伴黏液腺癌。结论MMR蛋白表达状态与结直肠癌患者年龄、肿瘤部位、分化程度、肿瘤分期和病理分型有关,结合RAS突变检测,可为结直肠癌患者的个体化治疗提供更多的依据。     

Slug regulates E‐cadherin expression in metastatic adenocarcinoma cells isolated from pleural fluid

Slug protein is a key regulator of epithelial‐mesenchymal transition, but its expression in cancer is less well studied. To evaluate the expression of slug, E‐cadherin and vimentin in adenocarcinoma cells from malignant pleural effusions, we analyzed 121 malignant pleural fluid specimens. Twenty‐eight nonmalignant pleural fluid specimens were analyzed as control. Besides clinical cytological diagnosis tests, immunofluorescence, immunocytochemistry and Western blotting methods were used. Results showed strong membrane staining of E‐cadherin in adenocarcinoma cells from pleural fluid. Slug mainly showed nucleus staining. Cytoplasma positive of vimentin was found in adenocarcinoma cells isolated from pleural fluid. Slug, E‐cadherin and vimentin expression was found in 43/121 (36%), 87/121 (72%) and 102/121 (84%) cases, respectively. Our data showed elevated levels of slug were accompanied by down regulation of E‐cadherin and the expression of vimentin in adenocarcinoma cells. In addition, there was no relationship between slug expression and patient's age or gender or tumor site. Hyperplasia epithelium cells from nonmalignant pleural fluid were uniformly negative for E‐cadherin and slug. In conclusion, the results demonstrated the inverse expression of slug and E‐cadherin in the majority of malignant pleural fluid cases compared with nonmalignant pleural fluid. The slug protein may be helpful to access the prognosis of patients with pleural fluid. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

P‐cadherin functional role is dependent on E‐cadherin cellular context: a proof of concept using the breast cancer model

P‐cadherin overexpression is associated with worse breast cancer survival, being a poor prognostic marker as well as a putative therapeutic target for the aggressive triple‐negative and basal‐like carcinomas (TNBCs). Previously, we have shown that P‐cadherin promotes breast cancer invasion of cells where membrane E‐cadherin was maintained; however, it suppresses invasion in models without endogenous cadherins, like melanomas. Here, we investigated if P‐cadherin expression would interfere with the normal adhesion complex and which were the cellular/molecular consequences, constituting, in this way, a new mechanism by which E‐cadherin invasive‐suppressor function was disrupted. Using breast TNBC models, we demonstrated, for the first time, that P‐cadherin co‐localizes with E‐cadherin, promoting cell invasion due to the disruption caused in the interaction between E‐cadherin and cytoplasmic catenins. P‐cadherin also induces cell migration and survival, modifying the expression profile of cells expressing wild‐type E‐cadherin and contributing to alter their cellular behaviour. Additionally, E‐ and P‐cadherin co‐expressing cells significantly enhanced in vivo tumour growth, compared with cells expressing only E‐ or only P‐cadherin. Finally, we still found that co‐expression of both molecules was significantly correlated with high‐grade breast carcinomas, biologically aggressive, and with poor patient survival, being a strong prognostic factor in this disease. Our results show a role for E‐ and P‐cadherin co‐expression in breast cancer progression and highlight the potential benefit of targeting P‐cadherin in the aggressive tumours expressing high levels of this protein. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Cytoplasmic β‐catenin accumulation is a good prognostic marker in upper and lower gastrointestinal adenocarcinomas

Norwood M G A, Bailey N, Nanji M, Gillies R S, Nicholson A, Ubhi S, Darnton J J, Steyn R S, Womack C, Hughes A, Hemingway D, Harrison R, Waters R & Jankowski J A
(2010) Histopathology 57, 101–111
Cytoplasmic β‐catenin accumulation is a good prognostic marker in upper and lower gastrointestinal adenocarcinomas Aims: β‐Catenin is an important molecule in cancer biology. Membranous β‐catenin enhances cellular differentiation and inhibits invasion by its action on E‐cadherin. The aim was to ascertain whether the cellular expression of these molecules in colorectal and oesophageal cancer specimens is associated with survival in patients with gastrointestinal cancer. Methods and results: Tumour samples from 149 patients undergoing resection for colorectal adenocarcinoma and 147 patients undergoing resection for oesophageal adenocarcinoma were retrospectively analysed using immunohistochemical techniques to assess β‐catenin expression. Increasing β‐catenin expression in the cytoplasm was associated with improved survival for colorectal cancer cases on both univariate (P = 0.003) and multivariate (P = 0.01) analysis. In addition, increased expression in the most recent cohort of oesophageal adenocarcinoma patients was associated with improved TNM staging (P = 0.007). Membrane expression was weakly associated with survival in colorectal cancer on univariate analysis (P = 0.09), but not on multivariate analysis (P = 0.21). Complete absence of β‐catenin expression at all three sites was associated with reduced 5‐year survival in colorectal cancer. Conclusions: This is one of the largest prognostic studies of β‐catenin in gastrointestinal adenocarcinoma. It shows that low levels of cytoplasmic β‐catenin expression are associated with reduced survival in patients with colorectal cancer as well as worse TNM staging in oesophageal adenocarcinoma (a recognized surrogate end‐point for survival). We believe this is the first time that this has been reported. This finding should be tested prospectively in oncological trials to validate whether the presence of cytoplasmic β‐catenin could be used as a prognostic marker for less aggressive disease.     

The prognostic significance of growth factors and growth factor receptors in gastric adenocarcinoma

We evaluated growth factors/receptors expression in gastric adenocarcinoma. Immunohistochemistry was used to evaluate epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), VEGF‐D, VEGF receptor (VEGFR)‐2, VEGFR‐3, transforming growth factor (TGF)‐α, TGF‐β1, and TGF‐β‐RII in tissue microarrays of adenocarcinoma, dysplasia, metaplasia, and gastritis. In adenocarcinoma, the expression rates of EGFR, VEGF, VEGF‐D, VEGFR‐2, VEGFR‐3, TGF‐α, TGF‐β1, and TGF‐β‐RII were 2.0%, 0%, 10.7%, 4.4%, 11.2%, 26.3%, 9.4%, and 19.5%, respectively. VEGF‐D, TGF‐α, TGF‐β1, and TGF‐β‐RII expression rate were higher in adenocarcinoma than in other groups. TGF‐β‐RII expression was correlated with VEGFR‐3, VEGF‐D, and TGF‐α expression in adenocarcinomas. Tumor location, histologic type, stage, lymphatic invasion, perineural invasion, angioinvasion, VEGF‐D, and VEGFR‐2 expressions were associated with patient survival in a log rank test and advanced stage and positive expression of VEGF‐D were poor prognostic factors using Cox analysis. VEGF‐D expression may be of prognostic value in gastric adenocarcinoma, whereas EGFR and TGF family expression may only have a minor influence.     

Mucinous carcinomas of the colon and rectum. An analysis of 62 stage B and C lesions.

The significance of a large extracellular mucinous component in colorectal adenocarcinomas continues to be controversial. We studied 62 stage B and C mucinous carcinomas defined by 60% or greater mucinous component. Sixty of these patients had matched nonmucinous adenocarcinoma controls. Patients with mucinous carcinomas with a residual adenoma had a significantly greater survival than those who did not. Overall, no difference was noted in 5-year survival between patients with mucinous carcinoma and nonmucinous adenocarcinoma (64% each). However, when the stages were considered separately, patients with stage B mucinous carcinoma fared significantly worse and had more local sites of treatment failure. We speculate that patients with stage B mucinous carcinoma may have a worse prognosis owing to the extra-cellular mucin, which may make a complete surgical extirpation more difficult.     

Cytokine expression associated with Helicobacter pylori and Epstein‐Barr virus infection in gastric carcinogenesis

Helicobacter pylori and Epstein‐Barr virus (EBV) infection, and associated cytokines are involved in gastric carcinogenesis. We investigated the expression of these cytokines and their relationship with clinicopathological characteristics. The study included specimens from 207 patients with gastric adenocarcinoma, 56 with chronic gastritis, 32 with metaplasia, and 30 with low‐grade epithelial dysplasia. Tissue microarrays were constructed and immunohistochemical staining for IL‐1β, IL‐6, IL‐10, IL‐17, p16, p21, TNF‐α, and TNFR1 was performed. EBV and H. pylori infection status was determined. IL‐1β, IL‐6, IL‐17, p16, and p21 protein expression was significantly higher in adenocarcinoma cases than in the other cases (p < 0.05). EBV was only noted in adenocarcinoma (13 cases, 6.3%). The H. pylori infection rate in adenocarcinoma was significantly higher than that in the other cases (p < 0.005). IL‐6 expression was associated with improved survival (p < 0.05), whereas IL‐17 expression was associated with decreased survival (p < 0.05). IL‐6 expression was inversely associated with angioinvasion, and disease stage (p < 0.05), whereas IL‐17 expression was associated with disease stage (p < 0.05). IL‐10 expression was correlated with IL‐1β and TNF‐α expression, and p16 expression was correlated with IL‐17 and EBV status. Our results indicate that IL‐6 and IL‐17 are associated with gastric carcinogenesis and may be considered prognostic factors.     

BMP‐2 Regulates the Formation of Oral Sulcus in Mouse Tongue by Altering the Balance Between TIMP‐1 and MMP‐13

The aim of this study is to investigate whether BMP‐2 regulates the oral sulcus formation of mouse embryonic tongue by modifying the expression of TIMP and MMP. The BMP‐2 siRNA induced a 180% increase in the depth of oral sulcus cavity (P < 0.01) by stimulating the invagination of oral sulcus into the mesenchymal tissues consisting of tongue floor, whereas the recombinant BMP‐2 suppressed the process in the organ culture system of mouse embryonic tongue. The BMP‐2 siRNA induced a 60% decrease in the expression of TIMP‐1 mRNA (P < 0.05) and a drastic decline in TIMP‐1 protein was observed around the oral sulcus in the BMP‐2 siRNA treated mandibles. The recombinant BMP‐2 induced a 220% increases in the expression of TIMP‐1 mRNA and the area of the immunostaining for TIMP‐1 around the oral sulcus was larger in the mandibles treated with the recombinant BMP‐2 than the vehicle. The BMP‐2 siRNA induced a 60% increase in the expression of MMP‐13 protein and a marked increase in the staining intensity for MMP‐13 was observed in the epithelial region of the BMP‐2 siRNA treated mandibles. The recombinant BMP‐2 induced a 70% decrease in the expression of MMP‐13 mRNA and the decrease was mainly observed in the tissues around oral sulcus. The expressions of BMP‐2, TIMP‐1, and MMP‐13 were verified in the tissues around in vivo developing oral sulcus at E11, 12, and 13 by immunohistochemistry. These results suggest that BMP‐2 regulates the formation of oral sulcus by altering the balance between TIMP‐1 and MMP‐13. Anat Rec 293:1408–1415, 2010. © 2010 Wiley‐Liss, Inc.