Potential Use of CTCs as Biomarkers in Renal Cancer Patients

We demonstrated that the CellCollector is an appropriate tool for detecting CTCs in RCC patients. We examined EpCAM and MUC1 expression levels in RCC tissues and cell lines and analyzed the detection rate of CTCs in blood samples ex vivo using an anti-EpCAM antibody-covered straight or spiraled CellCollector. Eight matched samples were examined for affinity to the anti-EpCAM vs. anti-EpCAM/anti-MUC1 antibody-covered wire. The use of this combination of antibodies allowed us to classify patients with lung metastasis. Finally, four patients were analyzed in vivo. In conclusion, both straight (ex vivo, in vivo) and spiraled (ex vivo) wires detected CTCs.

Evaluation of Ki67 Expression in Relation to Tumor Stage and Fuhrman Nuclear Grade of Renal Cell Carcinoma and MUC1 Expression in Clear Cell Renal Cell Carcinoma

BACKGROUND Renal cell carcinoma (RCC) is the most common malignant renal tumour in adults. Prognosis of RCC depends on various factors like tumour stage, nuclear grade and histological type. For planning adjuvant therapy, accurate prediction of prognosis is mandatory. In many studies, ki67 and MUC1 has shown to be of prognostic significance and immunohistochemical expression of these two markers plays an important role in determining the prognosis of RCC. The purpose of this study was to evaluate the Ki67 expression in histologically confirmed cases of RCC and MUC1 expression in clear cell renal cell carcinoma, and to correlate them with the stage and Fuhrman nuclear grade of the tumour, in order to determine their role as prognostic markers in RCC. METHODS This study was a retrospective study. A total of 50 specimens of renal cell carcinoma were studied. The specimens were total and partial nephrectomy done in the department of urology for a period of 3 years. Expression of Ki67 and MUC1 in RCC were studied by immunohistochemistry (IHC). Statistical analysis was performed and P value < 0.05 was considered significant. RESULTS Out of 50 RCC studied, Ki67 labelling index ≥ 15 % was found in 35 cases. For MUC1, immunoreactivity of more than 10 % of tumor cell was found in 28/34 of clear cell RCC. In this study, Ki67 labelling index showed statistically significant expression with the stage of tumor and the nuclear grade. MUC1 expression also showed significant correlation with nuclear grade and stage of clear cell RCC. CONCLUSIONS High Ki67 labelling index in renal cell carcinoma is seen to correlate with higher nuclear grade and stage of tumor. High level expression with circumferential staining pattern of MUC1 is seen in high grade tumours with increased risk of metastasis. So MUC1 and Ki67 can be considered as a marker of prognosis of RCC. KEY WORDS Renal Cell Carcinoma, Immunohistochemistry, Ki67, MUC1

Prognostic Value of MUC1 and its Correlation with Tumor-Infiltrating Immune Cells in Breast Cancer

Background: MUC1 is a transmembrane glycoprotein, aberrantly glycosylated and overexpressed in a variety of epithelial cancers, and plays a crucial role in cancer progression, especially in breast cancer. It is also an essential regulator for immune functionality, but the mechanisms whereby it effects immune infiltration in breast cancer remain uncertain. Methods: In this research, MUC1 expression was analyzed by the Oncomine and TIMER database. The association between MUC1 and prognosis was evaluated by Kaplan-Meier plotter database. The correlations of MUC1 with immune infiltration and immunological markers were assessed by TIMER database. Results: We found that MUC1 expression was significantly correlated with outcomes in multiple cancers, with the effect being particularly pronounced in breast cancer. Pathologically, elevated MUC1 expression was related with worse prognosis depending on intrinsic subtypes, ER status, patient stage, lymph node and TP53 mutation status in breast cancer. Specifically, low level of MUC1 seemed to be more favorable to luminal B patients with systemic treatment. MUC1 expression had significant negative correlations with infiltrating levels of CD8+ T cells, CD4+ T cells, B cells, macrophages, neutrophils, monocytes and dendritic cells (DCs) in breast cancer. Besides, MUC1 displayed strong regulations on macrophage polarization and diverse immunological gene markers. The patients with lower MUC1 and deeper immune infiltration predicted better prognosis. Conclusions: MUC1 is significantly associated with clinical prognosis and potentially plays an essential role in modulating cytotoxic T lymphocytes (CTLs), tumor associated macrophages (TAMs), natural killer cells (NK cells) and DCs in breast tumors. Collectively, MUC1 could be served as a valuable biomarker of predicting prognosis and immune infiltration for breast cancer patients.

Immunohistochemical Expression of HER2/neu, Ki-67 and MUC1 in Benign and Malignant Gall Bladder Lesions and its Association with Clinicopathological Parameters

Introduction: Gall Bladder Carcinoma (GBC) is a diagnostic and a therapeutic challenge. Although it is increasing, chronic cholecystitis remains the most worldwide gall bladder lesions, harbouring many epithelial changes that may end in carcinoma. Aim: To investigate the expression of HER2/neu (Human Epidermal Growth Factor Receptor 2), Ki-67 and MUC1 (Mucin 1) in malignant and non-malignant gall bladder lesions, and to evaluate its relation with clinicopathologic parameters of GBC. Materials and Methods: This retrospective study included 40 cases of GBC, eight cases of gall bladder dysplasia, 10 cases of gall bladder metaplastic changes and 25 cases of chronic cholecystitis as a control group. The blocks were collected from the Department of Pathology of Benha University Hospital, from January 2012 to December 2019. Immunohistochemical staining results of HER2/neu, Ki-67 and MUC1 were analysed and correlated by Statistical Package for the Social Sciences (SPSS) version 16 and Chi-square test or Fisher’s-exact tests. Results: Positive HER2/neu expression (+2, +3) was detected in 47.5% (19/40) of malignant cases and 12.5% (1/8) of dyspastic group, at the same time it was completely absent in the metaplastic and cholecystitis cases (p<0.01). Similarly, Ki-67 Labeling Index (LI) (≥20%) expression was found in 55% (22/40) of malignant group, while it was completely absent in the other three studied groups. All cases of malignant group 100% (40/40), 50% (4/8) of dysplastic one, one case of metaplastic (1/10) showed cytoplasmic expression of MUC1, at the same time it was completely absent in control group (0/25) (p<0.01). High MUC1 expression was found in 75% of both malignant (30/40) and dysplastic (6/8) studied cases and only one case (10%) of metaplastic group (p<0.01). There was a significant correlation between MUC1 expression and studied parameters of GBC. Conclusion: HER2/neu, and Ki-67 are overexpressed in GBC cases compared with control and dysplastic group. The study also highlights that MUC1 would be a better marker of malignant transformation of gall bladder epithelium and its depolarised expression would be reliable for detection of invasive carcinoma, so a new therapeutic agent can target these cell surface adhesion molecule (MUC1). HER2/neu can be considered as a candidate for targeted therapy in GBC treatment strategy.

The Expression Profile and Textural Characteristics of C595-Reactive MUC1 in Pancreatic Ductal Adenocarcinoma for Targeted Radionuclide Therapy

Improvements in the prognosis of pancreatic ductal adenocarcinoma (PDAC) rely on the development of effective treatments to target advanced disease. Mucin 1 (MUC1) is a transmembrane glycoprotein which is involved in the metastatic progression of PDAC and is a receptor-of-interest for targeted radionuclide therapy. The aim of this study was to determine the feasibility of MUC1-based targeted radionuclide therapy for PDAC, by evaluating the expression profile of MUC1 in different pancreatic cells and tissues using the C595 antibody. MUC1 expression was evaluated in four PDAC cell lines (PANC-1, BxPC-3, CAPAN-1 and AsPC-1) using flow cytometry and immunocytochemistry. Immunohistochemistry was performed on primary and metastatic PDAC, pancreatitis, pancreatic intra-epithelial neoplasia and normal pancreatic tissue samples to identify potential changes in C595-reactive MUC1 expression across different disease groups. C595-reactive MUC1 expression was found to varying degrees in the cell lines (11.5–93.1%). A pixel analysis of the immunohistochemical staining demonstrated highest MUC1 expression in primary PDAC tissue (mean pixel value of 205.4), followed by other pancreatic cancer types (204.9), pancreatic intra-epithelial neoplasia (203.8), metastatic PDAC (201.5), chronic pancreatitis (198.1) and normal pancreatic tissue (191.4). The increased expression in malignant tissues and reduced expression in benign tissues indicate that C595-reactive MUC1 is a potential target for targeted radionuclide therapy of PDAC.

GATA6 promotes epithelial-mesenchymal transition and metastasis through MUC1/β-catenin pathway in cholangiocarcinoma

GATA6 acts as an oncogene or tumour suppressor in different cancers. Previously, we found that aberrant expression of GATA6 promoted metastasis in cholangiocarcinoma (CCA). However, the mechanism by which GATA6 promotes metastasis in CCA is unclear. In the present study, we aimed to investigate the role of GATA6 in CCA cell epithelial–mesenchymal transition (EMT). Our results showed that GATA6 expression was positively associated with N-cadherin and vimentin expression but negatively associated with E-cadherin expression in 91 CCA samples. GATA6 promoted EMT and metastasis in CCA cells in vitro and in vivo based on knockdown and overexpression analyses. ChIP-sequencing data revealed that MUC1 is a novel downstream target of GATA6. GATA6 upregulated MUC1 expression through binding to both the 1584 and 1456 GATA-motifs in the promoter region and enhancing its transcription by luciferase reporter assays and point-mutant assays. MUC1 expression was positively associated with N-cadherin and vimentin expression but negatively associated with E-cadherin expression in 91 CCA samples. In addition, MUC1 promoted EMT in CCA cells based on knockdown and overexpression analyses. Moreover, MUC1 knockdown significantly abrogated the GATA6-induced EMT in CCA cells, indicating that MUC1 promoted EMT through upregulating MUC1 in CCA cells. β-Catenin is a putative transcriptional coactivator that regulates EMT in cancers. Our data showed that MUC1 expression was positively associated with nuclear β-catenin expression in 91 CCA samples. MUC1 upregulated nuclear β-catenin expression in CCA cells. Moreover, MUC1 bound to β-catenin in CCA cells based on protein immunoprecipitation analyses. MUC1 knockdown significantly decreased the binding of MUC1 to β-catenin, and thereby decreased nuclear β-catenin protein levels in CCA cells, indicating that MUC1 bound to β-catenin and increased its nuclear expression in CCA cells. Together, our results show that GATA6 promotes EMT through MUC1/β-catenin pathway in CCA, indicating potential implications for anti-metastatic therapy.

Smoking-associated increase in mucins 1 and 4 in human airways

Rationale Smoking-related chronic obstructive pulmonary disease (COPD) is associated with dysregulated production of mucus. Mucins (MUC) are important both for mucus secretion and epithelial defense. We have examined the distribution of MUC1 and MUC4 in the airway epithelial cells of never-smokers and smokers with and without COPD. Methods Mucosal biopsies and bronchial wash samples were obtained by bronchoscopy from age- and sex-matched COPD-patients (n = 38; GOLD I-II/A-B), healthy never-smokers (n = 40) and current smokers with normal lung function (n = 40) from the Karolinska COSMIC cohort (NCT02627872). Cell-specific expressions of MUC1, MUC4 and regulating factors, i.e., epithelial growth factor receptor (EGFR) 1 and 2, were analyzed by immunohistochemistry. Soluble MUC1 was measured by quantitative immunodetection on slot blot. Results The levels of cell-bound MUC1 expression in basal cells and in soluble MUC1 in bronchial wash were increased in smokers, regardless of airway obstruction. Patients with chronic bronchitis had higher MUC1 expression. The expression of MUC4 in cells with goblet cell phenotype was increased in smokers. The expression of EGFR2, but not that of EGFR1, was higher in never-smokers than in smokers. Conclusions Smoking history and the presence of chronic bronchitis, regardless of airway obstruction, affect both cellular and soluble MUC1 in human airways. Therefore, MUC1 may be a novel marker for smoking- associated airway disease.

Prognostic and Clinicopathological Significance of MUC Family Members in Colorectal Cancer: A Systematic Review and Meta-Analysis

Objective. To assess the association between MUC expression levels in colorectal cancer (CRC) tissues and prognosis and investigate the associations between MUC expression levels and CRC clinicopathological characteristics. Methods. The PubMed, Embase, Cochrane Library, and Web of Science databases were searched from inception through September 13, 2019, to identify studies investigating the association between MUC expression levels in CRC tissues and prognosis. Pooled hazard ratios (HRs) or odds ratio (ORs) with 95% confidence intervals (CIs) were used to evaluate associations between MUC expression levels and prognosis or clinicopathological characteristics, respectively. The heterogeneity between studies was assessed by the I2 values, whereas the likelihood of publication bias was assessed by Egger’s linear regression and Begg’s rank correlation test. Results. Among 33 included studies (n=6032 patients), there were no associations between combined MUC phenotype expression levels and overall survival (OS) or disease-free survival (DFS)/relapse-free survival (RFS) in patients with CRC. In subgroup analyses, the upregulated MUC1 expression (HR=1.50; 95% CI, 1.29–1.74; P<0.00001) was associated with poor OS. However, the upregulated MUC2 expression (HR=0.64; 95% CI, 0.52–0.79; P<0.00001) was associated with better OS. Furthermore, a high level of MUC1 expression (HR=1.99; 95% CI, 0.99–3.99; P=0.05) was associated with shorter DFS/RFS. However, patients with a low level of MUC2 tumors showed better DFS/RFS than patients with a high level of MUC2 tumors (HR=0.71; 95% CI, 0.49–1.04; P=0.08; P=0.0.009, I2=67%) and MUC5AC expression (HR=0.56; 95% CI, 0.38–0.82; P=0.003) was associated with longer DFS/RFS. In addition, a high level of MUC1 expression was associated with CRC in the rectum, deeper invasion, lymph node metastasis, distant metastasis, advanced tumor stage, and lymphatic invasion. A high level of MUC2 expression had a protective effect. High secretion of MUC5AC is associated with colon cancer compared with rectal cancer. Conclusion. The protein expression of MUC1 might be a poor biomarker in colorectal cancer and might play a role in tumor transformation and metastasis. However, the protein expression of MUC2 expression might have a protective effect. Furthermore, randomized controlled trials (RCTs) of large patients are needed to confirm the results.