E-cadherin Expression In Tumors Research Articles

E-cadherin expression in the tumor microenvironment of advanced epidermal growth factor receptor-mutant lung adenocarcinoma and the association with prognosis

BackgroundThe expression of programmed death-ligand 1 (PD-L1), tumor-infiltrating lymphocytes (TILs), E-cadherin, and vimentin in lung cancer tumor microenvironment is known to impact patient survival or response to therapy. The expression of these biomarkers may also differ between primary lung tumors and brain metastatic tumors. In this study, we investigated the interaction between these biomarkers in lung tumors with or without concomitant brain metastasis and the interaction with paired brain metastatic tumors.MethodsThe study included 48 patients with stage IV epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma. Sixteen of the forty-eight patients were diagnosed with brain metastasis, while the remaining thirty-two were not. All sixteen patients with brain metastasis had brain tumors. The expression of PD-L1, TILs (CD8+ T lymphocytes and FOXP3+ regulatory T lymphocytes), E-cadherin, and vimentin were evaluated using immunohistochemical (IHC) staining.ResultsPatients with brain metastasis exhibited a higher frequency of exon 19 deletion and uncommon EGFR mutations, a higher lung tumor vimentin score, worse progression-free survival (PFS), and overall survival (OS) than patients without brain metastasis. IHC staining showed no difference between paired lung and brain tumors. Patients with low PD-L1 expression had better PFS and OS. After multivariate analysis, higher body mass index, the presence of brain metastasis, bone metastasis, and uncommon EGFR mutations were correlated with worse PFS, while the presence of brain metastasis and high lung tumor E-cadherin score was associated with worse OS.ConclusionsIn patients with stage IV EGFR-mutant lung adenocarcinoma, high E-cadherin expression in the lung tumor might be associated with worse OS. Vimentin expression in the lung tumor was positively related to the risk of brain metastasis.

O218 E-CADHERIN EXPRESSION CORRELATES WITH PROGRESSION OF ESOPHAGEAL SQUAMOUS CELL CARCINOMA

Abstract Background & Aim E-cadherin is a member of the cadherin family and plays a major role in cell–cell adhesion of normal epithelium. Decreased or lack of E-cadherin expression has been reported in breast, colon and bladder cancer, and the disruption of the E-cadherin-mediated cell–cell adhesion is considered the first crucial step of metastasis. The esophageal stratified epithelial cells have many spines that intermingle with each other through desmosomes, in which E-cadherins play an important role for cell–cell adhesion. However, the significance of E-cadherin expression in the progression of esophageal squamous cell carcinoma (ESCC) still remains to be clarified. In the present study, we investigated the relation between E-cadherin expression in tumors and the clinicopathological features of ESCC. Method E-cadherin expression was evaluated in ESCC specimens of 61 patients who underwent surgery from April 2008 to December 2015 at the National Center for Global Health and Medicine by immunohistochemical staining. Distribution of E-cadherin in cancerous tissues was classified as Preserved (expression in cell membrane alone) or Disturbed (expression in cytoplasm or decreased/lack of expression of E-cadherin). Significance of the distribution pattern was analyzed by comparison to clinicopathological features. Results Thirty-seven of 61 (61%) cases revealed Preserved expression pattern while Disturbed distribution was observed in 24 of 61 (39%) cases. Compared to Preserved pattern, Disturbed expression of E-cadherin in tumors was significantly associated with advanced TNM stages (P=0.0462). Conclusion E-cadherin was expressed in most cases of ESCC; however, its distribution was not restricted in membrane but in the cytoplasm of tumors. Disturbed expression of E-cadherin in tumors correlated with advanced stage of ESCC, suggesting that the disruption of the E-cadherin-mediated cell–cell adhesion may play an important role in the progression of ESCC.

Clinical significance of altering epithelial-mesenchymal transition in metastatic lymph nodes of gastric cancer.

The E-cadherin, N-cadherin, and Snail genes are epithelial-mesenchymal transition (EMT)-inducible genes. Previous studies demonstrated that the expression of EMT markers in the primary tumor sites of gastric cancer correlates with tumor progression and prognosis. However, the clinical significance of the expression of these EMT markers in metastatic lymph nodes remains unclear. In the present study, we investigated the expression of these EMT markers in the primary tumor sites and metastatic lymph nodes. Immunohistochemistry was used to investigate the expression of E-cadherin, N-cadherin, and Snail in 89 primary tumors and 511 metastatic lymph nodes obtained from patients with gastric cancer. The weak expression of E-cadherin in tumors and lymph nodes increased with more lymph node metastasis and in more undifferentiated tumors. The strong expression of N-cadherin in lymph nodes correlated with more lymph nodes metastasis, an advanced stage, and poor prognosis. The weak expression of Snail in tumors correlated with lymphatic invasion. The strong expression of Snail in lymph nodes correlated with more lymph node metastasis and an advanced stage. The strong expression of Snail in tumors and its weak expression in lymph nodes correlated with more lymph node metastasis, an advanced stage, and poor prognosis. The expression of N-cadherin in metastatic lymph nodes is useful for predicting the prognosis of patients with gastric cancer. The Snail switch-namely, the positive-to-negative conversion of the Snail status-between primary tumors and lymph node metastasis may be important for confirming EMT and mesenchymal-epithelial transition.

SP-0015: The effects of radiotherapy on glioblastoma cell invasion

growth factors and tumor oxygenation collaborate to promote EMT and thereby contribute to radioresistance. Since within the tumor microenvironment, a plethora of signaling factors in conjunction with hypoxia may influence EMT induction, we also investigated the role of TGFβ addition or expression of the constitutive EGFRvIII mutant, stimuli known to influence EMT. We have previously shown that EGFRvIII contributes to increased malignancy and others have reported that EGFRvIII expression disrupts adherens junctions and that silencing EGFRvIII reduces expression of factors involved in EMT. Our data indicate that various stimuli emanating from tumor cells or their microenvironment can induce mesenchymal conversion of normal and cancerous epithelial cells. Interestingly, we show that combining these signals results in a more pronounced EMT-like induction. Moreover, EMT-like transformation by the microenvironment is reversible upon reoxygenation. We have correlated these findings with changes in the expression of E-cadherin, and found that cancer cells that express Ecadherin were more sensitive to radiation than their counterparts without E-cadherin. Thus changes in E-cadherin expression in tumors induced by changes in the microenvironment such as hypoxia and reoxygenation may contribute to the intrinsic sensitivity of tumor cells to radiotherapy. Interestingly, in addition to its influence on the process of EMT, our data provide evidence that EGFRvIII over expression promotes autophagy and survival of cancer cells under stress conditions such as hypoxia and starvation. Data on how these observations contribute to radiation resistance will be presented.

SP-0016: Head and neck cancer

growth factors and tumor oxygenation collaborate to promote EMT and thereby contribute to radioresistance. Since within the tumor microenvironment, a plethora of signaling factors in conjunction with hypoxia may influence EMT induction, we also investigated the role of TGFβ addition or expression of the constitutive EGFRvIII mutant, stimuli known to influence EMT. We have previously shown that EGFRvIII contributes to increased malignancy and others have reported that EGFRvIII expression disrupts adherens junctions and that silencing EGFRvIII reduces expression of factors involved in EMT. Our data indicate that various stimuli emanating from tumor cells or their microenvironment can induce mesenchymal conversion of normal and cancerous epithelial cells. Interestingly, we show that combining these signals results in a more pronounced EMT-like induction. Moreover, EMT-like transformation by the microenvironment is reversible upon reoxygenation. We have correlated these findings with changes in the expression of E-cadherin, and found that cancer cells that express Ecadherin were more sensitive to radiation than their counterparts without E-cadherin. Thus changes in E-cadherin expression in tumors induced by changes in the microenvironment such as hypoxia and reoxygenation may contribute to the intrinsic sensitivity of tumor cells to radiotherapy. Interestingly, in addition to its influence on the process of EMT, our data provide evidence that EGFRvIII over expression promotes autophagy and survival of cancer cells under stress conditions such as hypoxia and starvation. Data on how these observations contribute to radiation resistance will be presented.

N-cadherin Expression in Feline Mammary Tumors Is Associated With a Reduced E-cadherin Expression and the Presence of Regional Metastasis

The cadherin family of adhesion molecules regulates cell-cell interactions. N-cadherin is expressed by neural and fibroblast cells but not by normal epithelial cells. In human medicine, the role of N-cadherin in breast cancer remains controversial, but some studies have described the switch from E-cadherin to N-cadherin as a critical step in the malignant progression of neoplastic cells. The present study was carried out on 160 feline mammary tumors (21 adenomas and 139 carcinomas). The relationship between the immunohistochemical expression of N-cadherin in neoplastic epithelial cells and 2 established prognostic factors such as regional metastasis and tumor grade was examined. The results of the study showed a statistically significant relation between the expression of N-cadherin and the 2 prognostic factors, and also a reduced expression of E-cadherin in tumors that expressed N-cadherin.

Epithelial-to-Mesenchymal Transition Leads to Docetaxel Resistance in Prostate Cancer and Is Mediated by Reduced Expression of miR-200c and miR-205

Docetaxel is a standard chemotherapy for patients with metastatic prostate cancer. However, the response is rather limited and not all of the patients benefit from this treatment. To uncover key mechanisms of docetaxel insensitivity in prostate cancer, we have established docetaxel-resistant sublines. In this study, we report that docetaxel-resistant cells underwent an epithelial-to-mesenchymal transition during the selection process, leading to diminished E-cadherin levels and up-regulation of mesenchymal markers. Screening for key regulators of an epithelial phenotype revealed a significantly reduced expression of microRNA (miR)-200c and miR-205 in docetaxel-resistant cells. Transfection of either microRNA (miRNA) resulted in re-expression of E-cadherin. Functional assays confirmed reduced adhesive and increased invasive and migratory abilities. Furthermore, we detected an increased subpopulation with stem cell-like properties in resistant cells. Tissue microarray analysis revealed a reduced E-cadherin expression in tumors after neoadjuvant chemotherapy. Low E-cadherin levels could be linked to tumor relapse. The present study uncovers epithelial-to-mesenchymal transition as a hallmark of docetaxel resistance. Therefore, we suggest that this mechanism is at least in part responsible for chemotherapy failure, with implications for the development of novel therapeutics.

Randomized phase II trial of erlotinib (E) plus high-dose celecoxib (HD-C) or placebo (P) in advanced non-small cell lung cancer.

7518 Background: Cyclooxygenase-2 (COX-2)-dependent signalling represents a potential mechanism of resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) therapy in NSCLC. This is mediated in part through an EGFR-independent activation of MAPK/Erk by the COX-2 metabolite PGE2. In addition, PGE2 promotes downregulation of E-cadherin and epithelial to mesenchymal transition. We hypothesize that EGFR and COX-2 inhibition with E and HD-C will augment EGFR TKI efficacy by increasing tumor E-cadherin expression and blocking PGE2-mediated resistance to EGFR inhibition. Methods: Pts with stage IIIB/IV NSCLC who progressed following at least one line of therapy or refused standard chemotherapy were randomized to E (150mg/day)/HD-C (600mg/2x day) vs E/P in a 28-day cycle. Pts were stratified by smoking status and ECOG PS. The primary endpoint was PFS with 80% power to detect a 50% improvement; assessments were performed every 2 cycles. Secondary endpoints included response rate, OS and evaluation of molecular markers in tissue and serum to assess targeting COX-2-related pathways and evaluate EGFR TKI-resistance. All pts were required to have pre-treatment tissue available. Results: 107 pts were enrolled with comparable baseline characteristics in both arms. Disease control rate (DCR) was similar in both arms, and a trend toward improved PFS was seen in the E/HD-C arm with HR 0.81 (see Table). Analysis of those with EGFR wild type revealed a significantly increased PFS while those with EGFR mutation had similar PFS in both groups. Safety analysis showed similar toxicity in both arms. Additional biomarker correlations will be presented. Conclusions: The combination of E/C in metastatic NSCLC with HD-C is well tolerated and demonstrates significantly improved efficacy in EGFR wt population. This warrants further study into the COX-2-dependent mechanisms of primary resistance to EGFR TKI therapy. Supported by NIH 1P50 CA90388, K12 CA01727 and medical research funds from the Dept of Veterans’ Affairs. [Table: see text]

Triptolide inhibits ovarian cancer cell invasion by repression of matrix metalloproteinase 7 and 19 and upregulation of E-cadherin

Triptolide, a compound extracted from the traditional Chinese medicine preparation of Tripterygium wilfordii Hook F., has been reported to have anti-inflammatory and anti-cancer activities. However, its effect on ovarian cancer invasion is unknown. We observed that MMP7 and MMP19 expression increased in ovarian cancer tissue. Triptolide treatment inhibited the migration and invasion of ovarian cancer cells SKOV3 and A2780 at the concentration of 15 nM. We also observed that triptolide suppressed MMP7 and MMP19 promoter activity in a dose-dependent manner, down-regulating the expressions of these promoters on mRNA and protein level. Moreover, triptolide enhanced E-cadherin expression in ovarian cancer cells. In vivo, triptolide inhibited tumor formation and metastasis in nude mice, and suppressed MMP7 and MMP19 expression; it also enhanced E-cadherin expression in tumor in a dose-dependent manner. Over expression of MMP7 and MMP19, or suppression of E-cadherin expression partially abolished the inhibitory effect of triptolide on invasion of ovarian cancer cells. To summarize, triptolide significantly inhibited the migration and invasion of ovarian cancer cells by suppression of MMP7 and MMP19 and up-regulation of E-cadherin expression. This study shows that triptolide is a good candidate for the treatment of ovarian cancer and reduction of metastasis.

E-cadherin and β-catenin expression in sebaceous eyelid adenocarcinomas

Since the E-cadherin-catenin complex is of importance in the cancerogenesis of epithelial tumors, we examined the expression of E-cadherin and β-catenin in sebaceous adenocarcinomas of the eyelid and correlated the results with clinicopathological parameters. The study included histological specimen of 32 patients with sebaceous adenocarcinomas of the eyelid. The expression of E-cadherin, β-catenin, and Ki-67 nuclear antigen was analyzed by immunohistochemistry. The membranous expression of E-cadherin and of β-catenin was reduced in 17 (53%) patients and 18 (56%) patients, respectively, while elevated cytoplasmic levels and a nuclear accumulation of β-catenin were found in 27 (84%) and six (19%) patients, respectively. The low expression of membranous β-catenin was associated with a high degree of tumor infiltration (p = 0.02), high amount of inflammation (p = 0.01), and high degree of poor tumor differentiation (p < 0.001). In a similar manner, a low membranous E-cadherin expression in tumors was significantly associated with poor tumor differentiation (p = 0.001) and high tumor cell proliferation rate (p = 0.004). The Ki-67 proliferation index was significantly higher in the group with low membranous E-cadherin expression than in the group with high membranous E-cadherin expression (62.3 ± 20.8 versus 42.1 ± 15.3; p = 0.004). The membranous E-cadherin expression and the membranous β-catenin expression were significantly associated with each other (Fisher's exact probability test, p = 0.03). Sebaceous eyelid adenocarcinomas showed a decrease in the membranous expression of E-cadherin and of β-catenin. These changes were associated with poor tumor differentiation and an increase in the tumor infiltration and inflammation, pointing at a potential role of a low E-cadherin and low β-catenin expression for poor prognosis of sebaceous eyelid adenocarcinomas. Correspondingly, the cell proliferation index and the expression of E-cadherin were inversely correlated with each other. The findings suggest that the immunohistochemical detection of a low E-cadherin and low β-catenin expression may help in the examination and staging of sebaceous eyelid adenocarcinomas.

HOXA7 in Epithelial Ovarian Cancer: Interrelationships Between Differentiation and Clinical Features

Interrelationships between HOXA7 expression and ovarian cancer progression are investigated by cDNA array and by immunohistochemistry of normal ovaries and 538 epithelial ovarian tumor microarrays. Outcomes of patients and HOXA7 expression were compared by Kaplan-Meier survival curves. HOXA7 mRNA expression was higher in carcinomas than in benign tumors. HOXA7 protein was absent in normal surface epithelium but appeared in metaplastic regions. All carcinomas stained for HOXA7 protein, with staining intensities unrelated to histotype, grade, or patient outcome. There were significant associations of strong HOXA7 staining of stroma and tumor nuclei with the clear cell histotype (stroma: P = .0022, nuclei: P = .0003) and of weak/absent staining with serous carcinomas. Tumor E-cadherin expression correlated significantly with HOX7 staining in stroma (P = .0002) but not within tumors. HOXA7 staining of tumor cell nuclei is correlated significantly with improved disease-specific survival (P = .0104), which is suggestive of the biological and potentially clinical importance of subcellular HOXA7 localization.

E-cadherin and ?-catenin expression patterns in malignant melanoma assessed by image analysis

We investigated the expression of E-cadherin and beta-catenin in melanoma. Both proteins are components of adherens junctions but also play signalling roles in the wnt signal transduction pathway. Seventy malignant melanomas were analysed by immunohistochemistry and evaluated by image analysis as staining density, i.e. light permeability (LP). Comparison of mean values of relative LP for E-cadherin and beta-catenin in tumor tissue shows that levels of E-cadherin protein are significantly lower (259.67-116.23; t=22.7; p=0.000). The comparison of mean values of the relative LP of E-cadherin in melanoma to the LP in the adjacent normal skin also shows that the expression of E-cadherin in tumor is significantly lower (256.06-169.87; t=11.55, p=0.000). beta-catenin was observed in the cytoplasm in 30.6% of patients, in 24.2% in the cell membrane, in 21% in both the cytoplasm and membrane, in 1.6% in the membrane and nucleus and in 4.8% in the cytoplasm and nucleus, whereas in 17.7% of patients beta-catenin could not be observed. Patients with Clark 4 and 5 had significantly less beta-catenin than patients with Clark 2 and 3 (chi2=12.854; p=0.005). Changes in E-cadherin and beta-catenin levels have important roles in melanoma and could be used as molecular markers of disease progression.

The influence of pre-operative radiotherapy on the expression of p53 and adhesion molecules: correlation with treatment results in patients with squamous cell carcinoma or adenocarcinoma.

E-cadherin and the catenins are responsible for inter-cellular adhesion in epithelial tissues. E-cadherin and/or catenin expression is often altered in malignancies, leading to increased invasiveness and metastatic activity of tumour cells. Intact adhesion molecules reduce the risk on distant metastases. This is confirmed by studies mostly performed on surgical series, correlating e-cadherin expression in tumours with prognosis and treatment outcome. It has become more apparent that anti-cancer treatment by itself can also affect the expression of adhesion molecules. We therefore suggest that the prognostic value of e-cadherin expression may depend on the treatment modality and the sequence of therapies administered for malignant tumours. We used paraffin embedded specimens from patients with rectal tumours and patients with laryngeal tumours treated by short course radiotherapy before definitive surgery. Expression of p53, e-cadherin, and beta-catenin was determined in pre-radiotherapy biopsies and in the surgical specimens. Material was available from 37 patients. We found no correlation between the expression of p53, e-cadherin or beta-catenin and pre-treatment parameters. Mutated p53 in pre-radiation biopsy correlated with increased occurrence of distant metastases and there was an unexpected trend for abnormal e-cadherin expression to correlate with reduced metastases. The prognostic value of p53 no longer existed after examination of the surgical specimens (post-radiotherapy). There was a trend for e-cadherin to reverse from abnormal to normal expression in laryngeal squamous cell carcinomas after radiotherapy, in 5/7 cases it was accompanied with p53 conversion from positive to negative expression. Based on this study it is suggested that the predictive value of the e-cadherin expression for the occurrence of distant metastases in tumours treated by radiotherapy before surgery may be different from that found in tumours treated by surgery only. This may be related to the influence of radiotherapy on e-cadherin expression, especially in squamous cell carcinomas. Alteration in p53 expression was of predictive value only in pre-treatment biopsies and the beta-catenin status did not correlate with treatment outcome in this series.

Loss of E-cadherin expression associated with lymph node metastases in small breast carcinomas.

The National Breast Screening Programme affords the opportunity to study breast carcinomas at an early stage in their development. E-cadherin is a calcium-dependent, intercellular adhesion molecule whose loss of expression may facilitate the processes of invasion and metastasis of some human tumours. From a group of screen-detected ductal carcinomas less than or equal to 10 mm in diameter, 16 with lymph node metastasis were identified and matched for grade, size and patient age with node negative tumours. The level of expression of E-cadherin (detected by immunocytochemistry) was compared in the matched pairs using a simple semi-quantitative intensity distribution scoring system. The results showed a significant (P = 0.05 Wilcoxon paired rank test) reduction of E-cadherin expression in tumours with lymph node metastases compared to those without. In the context of the small size of these tumours it is proposed that these results support the hypothesis that reduction in E-cadherin expression is an early event in the development of metastases.