Negative E-cadherin Expression Research Articles

Characterization of E-Cadherin, SSEA-1, MSI-1, and SOX-2 Expression and Their Association with Pale Cells in Adenomyosis

Adenomyosis (AM) is a gynecological disease characterized by the invasion of endometrial glands and stroma within the myometrium. The etiology and pathogenesis of AM remain inadequately understood. Pale cells were identified as a novel cell type characterized by the absence of desmosomal contacts and light-colored cytoplasm. These cells were observed to migrate individually through ultra-micro ruptures in the basal membrane of the endometrial glands, translocating into the stroma and then further into the myometrium. Our study aimed to explore the possible stem cell properties of these pale cells. Forty hysterectomy specimens were analyzed using immunohistochemistry and immunofluorescence to assess negative E-cadherin expression and the positive expression of stem cell markers SSEA-1, MSI-1, and SOX-2. Immunohistochemical analysis revealed the presence of pale cells and occasionally rounded, enlarged E-cadherin-negative cells predominantly in the basal endometrial epithelium. The stem cell marker SSEA-1 was significantly elevated in the basalis epithelium, as well as in the ectopic epithelium. SSEA-1 positive cells were also identified in the stroma and myometrium. Sporadic colocalization of SSEA-1+/E-cadherin– cells was confirmed through immunofluorescence. The positive staining of pale cells for SSEA-1 and MSI-1 was also confirmed at the ultrastructural level by immunoelectron microscopy. These findings indicate that pale cells may possess stem cell characteristics, particularly a positive SSEA-1 profile, warranting further in vitro investigation into their role in the pathogenesis of adenomyosis.

The Expression of Ki-67 and E-cadherin in Triple Negative Breast Cancer: A Hospital Based Study

Introduction: Triple negative breast cancer (TNBC) is a subtype of breast cancer with aggressive nature as it does not respond to targeted chemotherapeutics. Expression of biomarkers is considered as an important factor for selecting treatment strategies and assessing prognosis. Objective: The aim of this study was to assess the expression of Ki-67 and E–cadherin in TNBC tissue. Methods: Immunohistochemical staining was done to assess the expression of proliferative marker Ki-67 and junctional complex marker E-cadherin in the Formalin Fixed Paraffin Embedded tissue (FFPE) of 80 TNBC patients. Ki-67 positive cells were identified on the basis of brown staining nuclei and E-cadherin expressed cells were considered positive on the basis of brown staining cell membrane. Ki-67 positive TNBCs were divided into low and high Ki-67 groups. Results: Ki-67 was expressed in 65% of the TNBC, the median of Ki-67 index was 30.25%. The difference between the medians of Ki-67 expression of low and high groups (7.5% and 53.25% respectively) was highly significant (p = 0.000). E-cadherin was negative in 67.5% of TNBC patients. The mean (±SD) age at diagnosis was 43.79 (±9.10) years. Most of the cancers were invasive ductal cell carcinoma (95%) and of grade II (70%). Lymph node metastasis was present in 73.8% patients and in 51% cases the cancer was in the left breast. Ki-67 expression was significantly associated with lymph node metastasis and E-cadherin expression was more negative in the left sided cancer. Conclusion: The study signifies that the TNBC has an early age at onset and Ki-67 expression is significantly high and associated with lymph node metastasis pointing the importance of this biomarker in TNBC patients. Negative expression of E-cadherin suggests close monitoring of the left sided TNBC.

Expression of Ki-67 and E-cadherin in patients with non-small cell lung cancer attending a tertiary care hospital

Background: E-cadherin and Ki-67 expressions may provide real-time insights into the tumor's status and can be utilized as targeted therapeutics for lung cancer. We aimed to explore the expression of Ki-67 and E-cadherin in non-small cell lung cancer (NSCLC) patients and to identify their association with clinicopathological features. Methods: In this cross-sectional study, forty formalin-fixed paraffin-embedded (FFPE) NSCLC tissue blocks were identified from January to October 2022, based on hospital records from the Department of Pathology of National Institute of Diseases of the Chest and Hospital, Dhaka Medical College and Hospital. Samples were reevaluated for tissue quality, diagnosis, and exclusion-inclusion criteria. Finally, twenty-five samples were analyzed, and relevant clinicopathological data were collected from patients or authorized representatives. Ki-67 and E-cadherin expression were analyzed by immunohistochemistry and their relationships with each other. Results: Ki-67 expression was positive in 40% of the NSCLC tissue, and E-cadherin was negative in 40% of the NSCLC tissue. No statistically significant relation was found between the expressions of Ki-67 and E-cadherin. No statistically significant association was found in Ki-67 and E-cadherin expressions with clinicopathological characteristics except E-cadherin with comorbidity. Conclusion: Positive expression of Ki-67 and negative expression of E-cadherin was found in two-fifths of NSCLC tissues but not significant. Simultaneous estimated of Ki-67 and E-cadherin may contribute to the treatment planning and predict prognosis of the NSCLC patients.

Negative E-cadherin expression on bone marrow myeloma cell membranes is associated with extramedullary disease.

Background: The loss of E-cadherin expression and the induction of N-cadherin are known as hallmarks of the epithelial-to-mesenchymal transition, an essential initial step in the process of metastasis in solid tumors. Although several studies have reported expressions of these cadherins in patients with multiple myeloma (MM), their clinical significance is unknown as MM cells are non-epithelial. Methods: In this study, we examined the expression of E- and N-cadherins by immunohistochemistry using bone marrow (BM) biopsy specimens from 31 newly diagnosed MM patients and in subsequent biopsy specimens from six of these. Results: Negative E-cadherin expression on BM myeloma cell membranes was significantly associated with the presence of soft-tissue masses arising from bone lesions and breaking through the cortical bone, referred to as extramedullary disease (EMD). Conclusions: Given the aggressive nature of EMD, our study suggests that screening for E-cadherin using BM immunohistochemistry is one measure that could predict the development of EMD in patients with MM.

The Correlation of E-cadherin Expression with Tumor Gradeand Tumor Budding in Endometrioid Endometrial Carcinoma

BackgroundEndometrial carcinoma is the fourth most common malignancy among women worldwide with increasing incidence and death rate every year. One of the types of endometrial carcinoma is endometrioid carcinoma, originated from atypical hyperplasia and develop into carcinoma. Lack of intercellular cohesiveness in the epithelial tumors such as endometrioid carcinoma occur due to lack of expression of E-cadherin. It can also causetumor invasion and metastatic through the epithelial-mesenchymal transition (EMT) process. Tumor differentiation and tumor budding are presumed to be histopathologic representations due to lack of cohesiveness and the occurrence of the EMT process so that these two things need to be related with the expression of E-cadherin on tumor cells.MethodsAn observational study was conducted using a cross-sectional approach with 46 cases of endometrioid endometrial carcinoma.Samples were obtained from 4 Anatomical Pathology Laboratories in West Sumatra during 2016-2019 in the form of paraffin blocks and HE slides that obtained from hysterectomy surgery. Reevaluation of tumor grade and tumor budding in HE slides was performed. E-cadherin expression in tumor cells was seen by immunohistochemical staining. Bivariate statistical analysis was performed using Fisher's Exact test and the results were considered significant if the p value was <0.05.ResultsSpecimens with negative E-cadherin expression were more common in grade 2 tumors (41.7%) and grade 3 tumors (50.0%) and in specimes with positive tumor budding (91.7%). Statistical analysis revealed a significant correlation between E-cadherin expression with tumor grade (p=0.000) and tumor budding (p=0.000).ConclusionExpression of E-cadherin has a significant correlation with the tumor grade and tumor budding in endometrioid endometrial carcinoma.

Особливості експресії молекул клітинної адгезії Е-кадгерину в різних молекулярних підтипах інвазивного протокового раку грудної залози

The purpose of the study was to determine the role of E-cadherin in the development and progression of molecular subtypes of invasive ductal breast cancer by assessing the expression of E-cadherin in various clinical and pathological prognostic parameters. Materials and methods. We demonstrated a comprehensive morphological, including immunohistochemical study of 193 cases of invasive ductal breast cancer with the molecular phenotype definition. General histological processing of samples was performed according to standard methods. Immunohistochemical studies for E-cadherin, ER, PR, c-erbB2, Ki-67 were performed according to standardized analytically validated protocols with the necessary controls. The grade of malignancy was determined according to the modified scheme of P. Scarff, H. Bloom and W. Richardson. E-cadherin expression level was quantified in 86 observations using the Qureshi counting system. Comparison of E-cadherin expression in different clinical and pathological parameters was evaluated using Pearson's test χ2. For all types of analysis, differences were considered significant at p <0.05. Results and discussion. E-cadherin expression in ductal breast cancer tissue is significantly lower in cases with lymph node metastases than without metastatic lymph node involvement (χ2 = 4.55, p = 0.031). Low expression of E-cadherin or its absence was associated with pT3 tumors, clinical stage 3, with G2 and G3 malignancies. Loss of E-cadherin expression has an unfavorable prognostic value. E-cadherin expression is associated with the molecular type of invasive breast ductal carcinoma. High E-cadherin expression was common in ER-positive tumors of the luminal A phenotype and was determined in patients of both premenopausal and postmenopausal age, suggesting that ER-positive expression may be involved in the regulation of E-cadherin expression. Low tumor activity of cells of invasive ductal breast cancer of the luminal subtype is accompanied by an increase in the adhesive properties of these cells due to the high level of expression of E-cadherin. E-cadherin is considered an independent marker of triple-negative breast cancer and is characterized by an unfavorable prognosis and short life expectancy. Triple-negative cancer was associated with a significant predominance of patients with low and negative E-cadherin expression (p = 0.011). Conclusion. Thus, E-cadherin is a potent tumor suppressor of breast cancer. According to this role in the progression of breast cancer, it was found that partial or complete loss of E-cadherin expression correlates with an unfavorable prognosis in patients

Negative E-cadherin expression on bone marrow myeloma cell membranes is associated with extramedullary disease.

Background: The loss of E-cadherin expression and the induction of N-cadherin are known as hallmarks of the epithelial-to-mesenchymal transition, an essential initial step in the process of metastasis in solid tumors. Although several studies have reported expressions of these cadherins in patients with multiple myeloma (MM), their clinical significance is unknown as MM cells are non-epithelial. Methods: In this study, we examined the expression of E- and N-cadherins by immunohistochemistry using bone marrow (BM) biopsy specimens from 31 newly diagnosed MM patients and in subsequent biopsy specimens from six of these. Results: Negative E-cadherin expression on BM myeloma cell membranes was significantly associated with the presence of soft-tissue masses arising from bone lesions and breaking through the cortical bone, referred to as extramedullary disease (EMD). Conclusions: Given the aggressive nature of EMD, our study suggests that screening for E-cadherin using BM immunohistochemistry is one measure that could predict the development of EMD in patients with MM.

Expression of Acute Phase Protein, Glial Fibrillary Acidic Protein, Epithelial Cadherin, and S100 Protein in Brain Tissues from Natural Cases of Ovine Encephalitic Listeriosis.

Listeriosis is a disease that is induced by infection with the Gram-positive bacterium Listeria monocytogenes. Much is still unknown about the pathogenesis of encephalitic listeriosis. We aimed to identify the contribution of glial fibrillary acidic protein (GFAP), epithelial cadherin (E-cadherin), S100, and acute-phase proteins (APPs) in pathogenesis, clinical and preclinical diagnosis of natural cases of encephalitic listeriosis. Of 1,325 Ossimi sheep, 64 were suspected of having listeriosis from 2018 to 2020. Prospective cohort evaluation of clinical and postmortem findings was performed, in addition to bacterial isolation, the measurement of APPs in serum and cerebrospinal fluid (CSF), and the histopathological and immunohistochemical evaluation of GFAP, S100, and E-cadherin. Infected sheep showed nervous symptoms ranging from neck stretching to complete paralysis. APPs were significantly increased in the CSF of both clinically and preclinically diseased animals; however, serum APPs were only significantly increased in clinically diseased animals. Histopathological evaluation revealed microabscesses, meningoencephalitis, and perivascular cuffing of the brainstem of infected sheep. Immunohistochemical investigations revealed strong expression of GFAP and S100 in necrotic areas and negative expression of E-cadherin. The measurement of CSF APPs could be useful in the preclinical diagnosis of sheep listeriosis. GFAP and S100 proteins could be involved in the pathogenesis of listeriosis; however, E-cadherin does not appear to be involved.

Suppression of circulating AP001429.1 long non-coding RNA in obese patients with breast cancer.

Long non-coding RNAs (lncRNAs), a type of cellular RNA, play a critical regulatory role in several physiological developments and pathological processes, such as tumorigenesis and tumor progression. Obesity is a risk factor for a number of serious health conditions, including breast cancer (BC). However, the underlying mechanisms behind the association between obesity and increased BC incidence and mortality remain unclear. Several studies have reported changes in lncRNA expression due to obesity and BC, independently encouraging further investigation of the relationship between the two in connection with lncRNAs. The present study was designed to first screen for the expression of 29 selected lncRNAs that showed a link to cancer or obesity in the blood of a selected cohort of 6 obese and 6 non-obese patients with BC. The expression levels of significantly expressed lncRNAs, AP001429.1, PCAT6, P5549, P19461 and P3134, were further investigated in a larger cohort of 69 patients with BC (36 obese and 33 non-obese), using reverse transcription-quantitative polymerase chain reaction. Results showed not only that AP001429.1 remained significantly downregulated in the larger cohort (P=0.002), but also that it was associated with several clinicopathological characteristics, such as negative HER2 status, negative E-cadherin expression, negative vascular invasion, negative margin invasion and LCIS. These findings suggest that obesity may have a role in inhibiting AP001429.1 expression, which may serve as a novel potential biomarker and therapeutic target for BC.

Analysis of the causes of unsuccessful hormonal treatment of non-atypical endometrial hyperplasia in premenopousal women

Annotation. Non-atypical endometrial hyperplasia (NEH), despite of benign morphology, may be insensitive to the hormonal treatment and has a tendency to recurrence and progression to atypia in some cases. The study purpose was the investigation of different type of progestins treatment results and comparison of progesterone receptors (PGR) and Е-cadherin expression in the sensitive NEH(+) and resistant NEH(–) to progestin treatment type of NEH. Prospective study of three groups of women with histologically confirmed NEH, who took different progestins during 6 months was done: І group – 96 women, who took micronized progesterone orally 200 mg per day continually, ІІ group – 161 women, who took dydrogesterone 20 mg per day continually, ІІІ group – 54 women, who were inserted LNG-IUD 52 mg. Control histopathological investigations of the endometrial samplings at 3 and 6 months were done. Expression of PGR and Е-cadherin by immunohistochemistry were investigated in the start samples of endometrium for all 63 NEH(–) women, 48 NEH(+) women and 20 control samples of normal proliferative and secretory endometrium. In the result of the study only nonsignificant and unreliable differences between different progestins efficacy were found. It was 75% normal endometrium samples till 6 months for micronized progesterone, 81.4% for dydrogesterone and 83.3% for LNG-IUD. Data analysis of PGR expression in the NEH(–) endometrium has shown significantly less week expression as for glandular cells (50.82±0.73), as for stromal cells (47.34±0.82) in comparison to the NEH(+) endometrial samples (glandular – 183.7±3.1; stroma 166.4±2.3; р<0.05) and normal proliferative (193.2±8.5 і 178.7±6.3 respectively; р<0.05) and secretory (140.2±4,4 і 116.6±3,1 respectively; р<0.05) endometrium. Е-cadherin expression in the glandular endometrial cells NEH(–) mostly was negative (86.4%) and 13.6% cells only demonstrated its week expression. NEH(+) women cells predominantly showed a positive reaction. It was often enough week (49.2%) and moderate (34.4%), but only in the 16.4% samples were negative. Thus, the use of progestogens for the treatment of NEH in women with low expression of PGR and negative expression of E-cadherin in the endometrium is inappropriate. Investigations of PGR та Е-cadherin expression in the endometrium of women with NEH before starting treatment may provide an opportunity to predict negative result in advance and chose alternative therapeutical approach.

Prognostic Value of Aquaporin-3, Vimentin and E-cadherin Expressions in Invasive Breast Carcinoma: An Immunohistochemical Study

Background: The preponderance of breast cancer-related deaths are the result of local invasion and distant metastasis; therefore, it is necessary to identify the factors underlying invasion and metastasis in order to develop novel treatment strategies and improve the survival of patients. In this regard, this study aimed to investigate the immunohistochemical expression and prognostic impact of aquaporin-3 (AQP3) and certain markers associated with epithelial-mesenchymal transition concerning invasive breast carcinoma of no special type. Method: Immunohistochemical expressions of AQP3, vimentin and E-cadherin were performed in 50 paraffin embedded specimens of such cases. We also assessed the relationship of their expressions with the clinicopathological variables and patients’ disease-free survival and overall survival. Results: There were significant associations between positive AQP3 and positive vimentin expressions and high tumor grade, large tumor size, lymph node metastasis, and advanced tumor stage. On other hand, negative E-cadherin expression had a significant correlation with high tumor grade, large tumor size, lymph node metastasis, distant metastasis, and advanced tumor stage. A significant association also existed between positive AQP3, positive vimentin and negative E-cadherin expressions and high tumor recurrence, short ‘three-year’ disease-free survival and overall survival. Conclusion: Positive AQP3, positive vimentin, and negative E-cadherin expressions are known as adverse prognostic markers and may predict survival in invasive breast carcinoma of no special type. It is proposed that AQP3 might play a role in breast cancer progression, invasion, and metastasis through induction of epithelial-mesenchymal transition.

An Integrative Morphomolecular Classification System of Gastric Carcinoma With Distinct Clinical Outcomes.

A robust morphomolecular classification system for gastric carcinoma is required. A 4-tier morphologic classification is proposed, including diffuse, intestinal, tubular, and lymphoid types. A tissue microarray for mismatch repair immunohistochemistry and Epstein-Barr virus (EBV) in situ hybridization were performed in 329 gastric carcinomas. DNA flow cytometry was used to detect aneuploidy in formalin-fixed paraffin-embedded samples. Lymphoid histology was the third most common histologic pattern at our institute and strongly associated with EBV infection and PMS2/MLH1-deficiency (both P<0.001). HER2 overexpression and SATB2 expression more frequently occurred in intestinal histology (both P<0.001). Loss of ARID1A expression was strikingly associated with lymphoid histology (P<0.001) and negative E-cadherin expression was correlated with diffuse histology (P=0.001). Programmed death-ligand 1 expression was most frequently present in lymphoid-type gastric carcinoma than other histologic subtypes and correlated with the molecular features of PMS2/MLH1-deficiency and EBV infection (all P<0.001). Aneuploidy was detected in 53% of gastric carcinomas and was highly correlated with intestinal type and the least with the lymphoid type (P<0.001). Notably, lymphoid-type gastric carcinoma showed the best outcome, whereas tubular type showed the worst survival rate (P<0.001). We integrated aneuploidy with morphologic patterns to propose a morphomolecular classification scheme, which served as a successful and independent prognostic factor in multivariate 5-year disease-free survival analysis (P<0.001). Overall, we describe an integrated morphomolecular classification system for gastric carcinomas to effectively predict patient outcomes. This system is cost-effective and reliable and can help select target therapeutics and facilitate clinical management.

Spinal versus intracranial meningioma: Expression of E-cadherin and Fascin with relation to clinicopathological features.

E-cadherin and Fascin are adhesive proteins that are expressed in many tumors. It was supposed that loss of expression of these proteins is associated with increased aggressiveness of the tumor. Whether spinal and intracranial meningiomas express adhesion proteins in different rates is not yet known. We aimed to investigate the expression of E-cadherin and Fascin in a large number of meningioma specimens and determine if clinical and prognostic significance existsMETHODS: One hundred and thirty-four spinal and intracranial meningioma samples were collected. Manual TMA blocks were constructed and immunohistochemistry for E-cadherin and Fascin was done. Focal or diffuse staining was considered positive. Intracranial meningioma occurred in significantly younger age than spinal ones. Most of spinal meningiomas were of transitional histology. E-cadherin was expressed in 38.8% of cases. Spinal meningiomas showed statistically significant negative expression of E-cadherin than intracranial tumors. All atypical meningiomas showed negative E-cadherin expression. Fascin was expressed in 9% of cases with significant expression in atypical cases. Aggressive behavior of meningioma could be explained in part by loss of E-cadherin and overexpression of Fascin especially in spinal meningiomas. Further studies are suggested to explore the biological aspects of spinal and intracranial meningiomas for constructing tailored targeted therapies.

Association of low expression of E-cadherin and β-catenin with the progression of early stage human squamous cervical cancer.

The precise involvement and mechanisms of human papilloma virus type 16 (HPV16) in epithelial-mesenchymal transition (EMT) of cervical intraepithelial neoplasia (CIN) and squamous cervical cancer (SCC) remain unknown. The present study aimed to examine the expression of EMT indicators and their association with HPV16 in CIN and early stage SCC, and their prognostic value in early stage SCC. The expression levels of E-cadherin, N-cadherin, β-catenin, vimentin, and fibronectin were determined by immunohistochemistry in 40 patients with normal uterine cervix, 22 patients with CIN1, 60 patients with CIN2-3, and 86 patients with SCC, stage Ia-IIa, according to the International Federation of Gynecology and Obstetrics. The expression of the epithelial indicators E-cadherin and β-catenin gradually declined, and the mesenchymal indicators N-cadherin, vimentin, and fibronectin increased with progression of the cervical lesions (P<0.05). Patients with SCC with lymph node metastasis, parametrial invasion, negative E-cadherin, and negative β-catenin expression had shorter overall survival (P=0.001, P=0.015, P=0.014, and P=0.043, respectively) and disease-free survival (P=0.002, P=0.021, P=0.025, and P=0.045, respectively) time. Multivariate survival analysis indicated that lymph node metastasis [Hazard ratio (HR)=3.544; P=0.010], parametrial invasion (HR=2.014; P=0.007) and E-cadherin expression (HR=0.163; P<0.001) were independently associated with overall survival, but also with disease-free survival (HR=3.612, P=0.009; HR=1.935, P=0.011; HR=0.168, P<0.001, respectively). In patients with CINs, HPV16 infection was negatively correlated with the expression of E-cadherin, and positively correlated with the expression of N-cadherin, vimentin, and fibronectin. EMT occurs during the progression of CINs to early stage SCC, and is associated with HPV16 infection in CINs. Lymph node metastasis and parametrial invasion are poor prognostic factors for SCC, while positive E-cadherin expression may serve as a protective prognostic factor for SCC.

Metadherin overexpression in perihilar cholangiocarcinoma is associated with lymph node metastasis and poor prognosis.

Metadherin (MTDH) is a protein that is also named astrocyte elevated gene-1, and is highly expressed in a number of different tumor tissues. Although the expression of MTDH is associated with tumor invasion and recurrence, the expression of this protein in perihilar cholangiocarcinoma (PCCA) and its clinical use have not yet been investigated. In the present study, the expression of MTDH in patients with PCCA was investigated in order to determine its clinicopathological use. An immunohistochemical method was used to detect MTDH expression and the epithelial-mesenchymal transition markers E-cadherin and vimentin in 66 cases of PCCA. In addition to the expression of MTDH, the clinical and pathological data and the postoperative outcomes were analyzed. The MTDH positive expression rate was 48.5% (32/66) in PCCA. A significantly higher MTDH expression level was identified in the poor tumor differentiation group compared with the well differentiation group (P=0.007). In the positive lymph node metastasis group, a significantly higher MTDH expression level was revealed compared with the negative lymph node metastasis group (P=0.023). No association was noted with regard to the expression of MTDH and the variables age, sex, tumor diameter, tumor grade and tumor classification stage. Positive MTDH expression was significantly associated with high vimentin expression (P=0.037) compared with negative vimentin expression and inversely associated with positive E-cadherin expression compared with negative E-cadherin expression (P=0.030). Survival analysis suggested that the high MTDH expression group was associated with a worse overall survival (OS) rate and recurrence free survival (RFS) rate compared with the low MTDH expression group (P<0.001 and P=0.01, respectively). Cox regression analysis indicated that the Tumor-Node-Metastasis, surgery margin and high MTDH expression were independent OS and RFS factors for PCCA. MTDH expression may serve an important function in PCCA tumor growth and metastasis. Targeting MTDH may have important therapeutic applications for patients with PCCA.

High expression of E-cadherin and Ki-67 associated with functional/dysfunctional phenotypes of tumor-infiltrating lymphocytes among Chinese patients with operable breast cancer.

ObjectiveBreast cancer has become the most common cancer in women in China, and the clinicopathological features differ from those in Western patients. This study was performed to investigate the distribution of programmed cell death protein 1 (PD-1)+/PD-1− tumor-infiltrating lymphocytes (TILs) and its association with clinicopathological features among Chinese patients with breast cancer.MethodsIn total, 133 consecutive patients with primary breast cancer were recruited into this cross-sectional study from 2012 to 2013. TILs were measured by cell counts under high-power fields (HPFs). Immunohistochemistry was used to detect PD-1 expression on tumor-infiltrating lymphocytes in the microenvironment.ResultsThe median cell counts of the overall TILs, PD-1+ TILs, and PD-1− TILs were 80, 18, and 55/HPF, respectively. The number of PD-1− TILs was significantly lower in older than younger patients (50 vs. 60/HPF). Patients with positive E-cadherin expression had more PD-1− TILs than patients with negative E-cadherin expression (57 vs. 27/HPF). The Ki-67 index was positively correlated with the cell counts of PD-1+ TILs, and the correlation coefficient was 0.29.ConclusionsPD-1 expression on TILs had different clinicopathological features in Chinese patients with breast cancer. E-Cadherin expression was associated with PD-1− TILs; however, Ki-67 expression was associated with PD-1+ TILs.

Clinicopathological and prognostic significance of Ras association and pleckstrin homology domains 1 (RAPH1) in breast cancer.

Ras association and pleckstrin homology domains 1 (RAPH1) is involved in cytoskeleton regulation and re-epithelialisation in invasive carcinoma and, therefore, may play a key role in carcinogenesis and metastasis. We, herein, investigated the biological and clinical significance of RAPH1 in breast cancer using large annotated cohorts. The clinicopathological and prognostic significance of RAPH1 was assessed at the genomic and transcriptomic levels using The Cancer Genome Atlas (TCGA) dataset (n = 1039) and the results were validated using the Molecular taxonomy of breast cancer international consortium (METABRIC) cohort (n = 1980). RAPH1 protein expression was evaluated by immunohistochemistry in a large, well-characterised cohort of early-stage breast cancer (n = 1040). In both the TCGA and METABRIC cohorts, RAPH1 mRNA expression and RAPH1 copy number alteration were strongly correlated. RAPH1 mRNA overexpression was significantly correlated with high expression of adhesion and EMT markers including CDH1, TGFβ1 and CD44. RAPH1 mRNA overexpression was a significant predictor of a poor prognosis (Hazard ratio 3.88; p = 0.049). High RAPH1 protein expression was associated with higher grade tumours with high proliferation index, triple negative phenotype and high E-cadherin expression. High RAPH1 protein expression was an independent predictor of shorter survival (Hazard ratio 4.37; p = 0.037). High RAPH1 expression is correlated with aggressive breast cancer phenotypes and provides independent prognostic value in invasive breast cancer.

Expression of PD-1 on CD4+ Tumor-Infiltrating Lymphocytes in Tumor Microenvironment Associated with Pathological Characteristics of Breast Cancer.

Objective This study aimed to investigate the correlation of CD4+/PD-1+ or CD4+/PD-1− tumor-infiltrating lymphocytes with pathological characteristics in breast cancer patients. Methods A cross-sectional study consecutively recruited 133 patients with invasive ductal breast cancer. The expression of CD4, programmed cell death protein 1 (PD-1), CK7, CK20, E-cadherin, or Ki-67 was detected by immunohistochemistry. The associations between CD4+/PD-1+ or CD4+/PD-1− tumor-infiltrating lymphocytes and pathological characteristics were evaluated. Results Elderly patients intended to have a lower level of CD4+/PD-1− tumor-infiltrating lymphocytes (p < 0.05). Patients with positive E-cadherin expression had higher median cell counts of CD4+/PD-1− tumor-infiltrating lymphocytes than patients with negative E-cadherin expression (30/HPF versus 10/HPF, p < 0.05). Counts of CD4+/PD-1+ tumor-infiltrating lymphocytes had a significant correlation with Ki-67 index that the correlation coefficient was 0.29 (p = 0.001). Positive CK20 expression was related to a higher level of CD4+/PD-1− tumor-infiltrating lymphocytes than negative CK20 expression (73/HPF versus 30/HPF, p < 0.05). Conclusion CD4+/PD-1+ or CD4+/PD-1− tumor-infiltrating lymphocytes showed diverse association with pathological features of breast cancer. CD4+/PD-1+ tumor-infiltrating lymphocytes had a significant relationship with Ki-67 expression whereas CD4+/PD-1− tumor-infiltrating lymphocytes had a significant relationship with E-cadherin expression. Further studies are warranted to explore the immunomodulatory effects of phenotypes of CD4+ T cell subsets in breast cancer.

Role of epithelial-mesenchymal transition markers E-cadherin, N-cadherin, β-catenin and ZEB2 in laryngeal squamous cell carcinoma.

Epithelial-mesenchymal transition (EMT) allows neoplastic cells to gain the invasive phenotype and become migratory, which is required for cancer progression and metastasis. In the present study, the expression of EMT-associated biomarkers and their association with clinicopathological parameters in laryngeal squamous cell carcinoma (LSCC) was investigated. E-cadherin, N-cadherin, β-catenin and zinc finger E-box binding homeobox 2 (ZEB2) protein expression was evaluated with immunohistochemistry in a cohort of 76 patients with operable LSCC. The association between these transition markers, clinicopathological parameters and their prognostic impact in LSCC was analyzed. Immunohistochemical analysis revealed that EMT-associated proteins were differentially expressed between LSCC and adjacent non-neoplastic laryngeal tissue. Negative E-cadherin expression and positive N-cadherin, β-catenin and ZEB2 expression were associated with a later tumor (T) stage, decreasing tumor differentiation and a reduced overall survival (OS) time (OS: E-cadherin, P=0.016; N-cadherin, P=0.003; β-catenin, P=0.002; ZEB2, P=0.0003). E-cadherin/β-catenin co-expression was significantly associated with the majority of clinicopathological parameters assessed, including lymph node metastases, T stage and tumor cell differentiation (P=0.004, P=0.005, and P<0.001, respectively). Multivariate analysis indicated that T stage and the positive expression of β-catenin and ZEB2 were independent risk factors for OS in LSCC (P=0.014, P=0.025 and P=0.003, respectively). It was concluded that EMT mediates tumor progression, and reduces OS time in patients with LSCC. E-cadherin/β-catenin co-expression may be associated with clinicopathological parameters. T stage, and the positive co-expression of β-catenin and ZEB2 may be independent predictors of prognosis in LSCC.

P2Y2 receptor promotes the migration and invasion of breast cancer cells via EMT-related genes Snail and E-cadherin.

Adenosine 5′-triphosphate (ATP) is one of the most abundant biochemical constituents within the tumor microenvironment and is postulated to play critical roles in the progression of a number of types of tumors via interaction with the P2Y2 receptor. In the present study, we demonstrated that the P2Y2 receptor was highly expressed in MCF7 and Hs578T breast cancer cells. Downregulation of the P2Y2 receptor by small interfering RNA (siRNA) significantly attenuated ATP- or UTP-driven migration and invasion of the breast cancer cells as well as expression of EMT-related genes Snail and E-cadherin. Consistent with the observations in vitro, the P2Y2 receptor was found to be abundantly expressed at the invasive edge of the tumor, in infiltrating tumor cells in breast adipose tissues and/or the cancer embolus in the lymphatic sinuses compared with the tumor core areas. Furthermore, high Snail expression and weak or negative expression of E-cadherin were observed at the invasive edge of tumors. Taken together, these data indicate that the P2Y2 receptor promoted cell migration and invasion in breast cancer cells via EMT-related genes Snail and E-cadherin.