The roles of epidermal growth factor receptor in viral infections.

Viruses are intracellular pathogen that exploit host cellular machinery for their propagation. Extensive research on virus-host interaction have shed light on an alternative antiviral strategy that targets host cell factors. Epidermal growth factor receptor (EGFR) is a versatile signal transducer that is involved in a range of cellular processes. Numerous studies have revealed how viruses exploit the function of EGFR in different stages of viral life cycle. In general, viruses attach onto the host cell surface and interacts with EGFR to facilitate viral entry, viral replication and spread as well as evasion from host immunosurveillance. Moreover, virus-induced activation of EGFR signalling is associated with mucin expression, tissue damage and carcinogenesis that contribute to serious complications. Herein, we review our current understanding of roles of EGFR in viral infection and its potential as therapeutic target in managing viral infection. We also discuss the available EGFR-targeted therapies and their limitations.

Echinostoma caproni (Trematoda): differential in vivo mucin expression and glycosylation in high- and low-compatible hosts.

Enhanced mucus production and release appears to be a common mechanism for the clearance of intestinal helminths, and this expulsion is normally mediated by Th2-type immune responses. To investigate the factors determining the expulsion of intestinal helminths, we have analysed in vivo expression of mucin genes at the site of infection in two host species displaying different compatibility with Echinostoma caproni (Trematoda). Surprisingly, a general down-regulation on mucin mRNA expression was detected in low-compatible hosts (rats) coinciding with the development of Th2/Th17 responses and the early rejection of the worms from the intestinal lumen. This suggests the existence of a mechanism by which the parasites can modulate the mucus barrier to favour their survival. In highly compatible hosts (mice), some mucin genes were found to be up-regulated throughout the infection, probably, to protect the intestinal epithelium against the infection-induced inflammation developed in this host species. Moreover, infection-induced changes on mucin glycans were also studied by lectin histochemistry. Similar alterations were detected in the ileum of infected mice and rats, except with SNA lectin, indicating that sylated mucins might play an important role in determining the evolution of the infection in each host species.

Topical Adenosine Inhibits Inflammation and Mucus Production in Viral Acute Rhinosinusitis.

OBJECTIVE: Viral acute rhinosinusitis (ARS) is the leading cause of work and school absence and antibiotic over-prescription. There are limited treatment options available to ameliorate the symptoms caused by viral ARS. We have previously demonstrated that topical adenosine treatment enhances mucociliary clearance in the sino-nasal tract. Here, we assessed the therapeutic potential of topical adenosine in a mouse model of viral ARS. METHODS: The effect of topical adenosine on inflammatory response and mucin gene expression was examined in a mouse model of viral ARS induced by respiratory syncytial virus (RSV) nasal-only infection. We also investigated the inflammatory effect of both endogenous and exogenous adenosine in the sino-nasal tract. RESULTS: Topical adenosine significantly inhibited the expression of pro-inflammatory cytokines, goblet hyperplasia, mucin expression, and cell damage in the nose of mice with viral ARS. This treatment did not prolong virus clearance. This inhibitory effect was primarily mediated by the A2A adenosine receptor (AR). Although previous studies have shown that adenosine induces a robust inflammatory response in the lungs, neither endogenous nor exogenous adenosine produced inflammation in the sino-nasal tract. Instead, exogenous adenosine inhibited the baseline expression of TNF and IL-1ß in the nose. Additionally, baseline expression of ARs was lower in the nose than that in the trachea and lungs. CONCLUSION: We demonstrated that intranasal adenosine administration effectively decreased inflammation and mucus production in a mouse model of viral ARS. LEVEL OF EVIDENCE: N/A Laryngoscope, 133:2095-2103, 2023.

Nanosecond pulsed electric fields impair viability and mucin expression in mucinous colorectal carcinoma cell.

Nanosecond pulsed electric fields (nsPEFs) are a non-thermal technology that can induce a myriad of biological responses and changes in cellular physiology. nsPEFs have gained significant attention as a novel cancer therapy. However, studies investigating the application of nsPEF in mucinous carcinomas are scarce. In this study, we explored several biological responses in two mucinous colorectal adenocarcinoma cell lines, LS 174T and HT-29, to nsPEF treatment. We determined the overall cell survival and viability rates following nsPEF treatment using CCK-8 and colony formation assays. We measured the intracellular effects of nsPEF treatment by analyzing cell cycle distribution, cell apoptosis and mitochondrial potential. We also analyzed mucin production at both mRNA and protein levels. Our results showed that nsPEF treatment significantly reduced mucinous cell viability in a dose-dependent manner. nsPEF treatment increased cell cycles arrest at G0/G1 while the proportion of G2/M cells gradually decreased. Cell apoptosis increased following nsPEF treatment with a clear loss in mitochondrial membrane potential. Furthermore, the protein expression of functional mucin family members decreased after nsPEF treatment. In conclusion, nsPEF treatment reduced MCRC cell viability, cell proliferation, and mucin protein production while promoted apoptosis. Our work is a pilot study that projects some insights into the potential clinical applications of nsPEFs in treating mucinous colorectal carcinoma.

Expression of Mucins in Salivary Gland Mucoepidermoid Carcinoma.

Mucoepidermoid carcinoma (MEC) is the most common malignant salivary gland tumour in both adults and children. Histological grading of MEC is subjective, but plays an important role in predicting patient prognosis. Epithelial mucin (MUC) status may aid in establishing a more accurate grade. This study aimed to investigate the expression of various mucins (MUC1, MUC2, MUC4 and MUC5AC) in MECs to determine a possible correlation with tumour grade. Fifteen cases of each tumour grade (low-, intermediate-, and high-grade) were retrieved from the pathology archives of the Department of Oral Pathology and Oral Biology at the University of Pretoria. The patients included 23 men and 22 women, and ranged from 13 to 85 years (mean 49.8 years). Sections from formalin-fixed paraffin-embedded (FFPE) tissue were used for fluorescence in situ hybridization (FISH) for MAML2 rearrangements and MUC immunohistochemical analysis. The percentage immunohistochemical expression of the neoplastic mucous cells was evaluated first, followed by the overall percentage expression of all tumour cells. The results indicated that MUC1 overexpression may be a reliable marker of high-grade MECs, whereas MUC4 overexpression may be more indicative of low-grade tumours. MUC5AC expression was considered an unreliable marker in determining grade. MUC2 was only expressed in a single case of MEC and may be considered a useful marker to exclude MEC as a diagnostic possibility. This study demonstrates that MECs show an altered MUC expression pattern that can be used for diagnostic purposes and to aid in establishing a more accurate tumour grade.

Food-grade titanium dioxide (E171) induces anxiety, adenomas in colon and goblet cells hyperplasia in a regular diet model and microvesicular steatosis in a high fat diet model.

Food-grade titanium dioxide (E171) is a white additive widely used in solid and liquid food products. There is still debate about E171 toxic effects after oral consumption since this additive is deposited in colon, liver, spleen, testis and brain. The consumption of E171 commonly occurs with Western diets that are characterized by a high fat content. Thus, E171 could worsen adverse effects associated with a high fat diet (HFD) such as anxiety, colon diseases and testicular damage. We aimed to evaluate the effects of E171 on anxiety-like behavior, colon, liver and testis and to analyze if the administration of a HFD could exacerbate adverse effects. E171 was administered at ~5 mg/kgbw by drinking water for 16 weeks and mice were fed with a Regular Diet or a HFD. E171 promoted anxiety, induced adenomas in colon, goblet cells hypertrophy and hyperplasia and mucins overexpression, but had no toxic effects on testicular tissue or spermatozoa in regular diet fed-mice. Additionally, E171 promoted microvesicular steatosis in liver in HFD fed-mice and the only HFD administration decreased the spermatozoa concentration and motility. In conclusion, E171 administration increases the number of adenomas in colon, induces hypertrophy and hyperplasia in goblet cells and microvesicular steatosis.

Immunohistochemical expression profiles of MUC1 and MUC2 mucins in urothelial tumors of bladder.

BACKGROUND: Mucins may show aberrant expression, localization, and/or glycosylation in multiple malignancies. However, information regarding expression of these mucins is mostly unknown in urothelial tumors. AIM: This study was conducted for examining the expressions of membrane associated and secreted mucin (MUC1) and a secreted gel-forming mucin (MUC2) in urothelial tumors of the urinary bladder. SUBJECTS AND METHODS: Archival transurethral resection materials of 97 urothelial carcinoma cases were reexamined light microscopically and graded according to the 2004 WHO Classification. Pathological stage was given as pTa, pT1, and pT2. Demonstrative sections were recut for immunohistochemistry for MUC1 and MUC2. The results were statistically analyzed, and P < 0.05 was considered statistically significant. RESULTS: The positivity for MUC1 and MUC2 was 89.7% and 44.3%, respectively. Independent from pathological stage of the tumor, MUC1 expression showed statistically significant correlation with tumor grade (P < 0.05). We did not find any correlation between pathological stage and MUC1 and MUC2 expression (P > 0.05). MUC1 staining pattern in papillary urothelial neoplasm of low malignant potential cases was more commonly apical and superficial (luminal cell layer only). Intermediate cells ± basal cells or isolated cells or islands of tumor cells with cytoplasmic and/or circumferential membrane positivity for MUC1 and MUC2 were more commonly observed in both low- and high-grade carcinomas. The difference between groups in terms of MUC1 and MUC2 staining was statistically significant (P < 0.05). CONCLUSIONS: The staining patterns of both mucins are different between urothelial papillary tumors and may be used to make a differentiation, especially for low-grade papillary urothelial lesions. This difference may also be important in the carcinomatous transformation of urothelial neoplastic and preneoplastic lesions.

Diagnostic and prognostic impact of mucin 1-6 expression in non-small cell lung cancer.

BACKGROUND: The prognostic significance and clinico-pathological characterization of mucin (MUC) expression in non-small cell lung cancer (NSCLC) is controversial and little studied. AIMS: This study aims at elucidating this issue on the largest and most detailed cohort so far. SETTINGS AND DESIGN: We examined the expression of MUC 1, 2, 4, 5AC and 6 on 371, well documented, surgically resected NSCLC cases. MATERIALS AND METHODS: Immunohistochemical results were correlated with several of our previously studied, relevant parameters on this cohort including a follow-up period of up to 20 years. An additional point we examined for practical reasons that has not been addressed so far, was the possible assistance of MUC expression for the differentiation between a primary lung adenocarcinoma and metastasis from a known pancreatobiliary primary tumor. STATISTICAL ANALYSIS USED: Cronbach's Alpha reliability correlation, Spearman's correlation, ANOVA means of comparison with additional Kruskall-Wallis H-test, and Kaplan-Meier survival analysis were employed as statistical analyses in this study. RESULTS AND CONCLUSIONS: MUCs were associated with histologic subtypes, tumor differentiation and members of the epidermal growth factor receptor signaling pathway, although they were not found to be significant for prognosis. Expression of MUC1 correlated with certain other markers and may point to a group of patients relevant for upcoming treatment strategies involving MUC1. According to our findings, we also recommend additional MUC5AC staining for a thyroid transcription factor 1-negative adenocarinoma in the lung for the differentiation of a possible metastasis in the presence of a pancreatic ductal adenocarcinoma.

Immunohistochemical Study of MUC1, MUC2 and MUC5AC Expression in Primary Breast Carcinoma.

INTRODUCTION: Breast Cancer (BC) is the second most common cancer among women in India and accounts for 7% of global burden of BC and one-fifth of all Cancers (CA) among women in India. AIM: This study was conducted for studying the expression of MUC1, MUC2 and MUC5AC in breast carcinoma. MATERIALS AND METHODS: Fifty cases of primary breast carcinoma diagnosed between years 2013 to 2015 were included in the study. Manual tissue array technique was applied for cases subjected to Immunohistochemistry (IHC). An analysis of the expression of IHC markers (MUC1, MUC2, MUC5AC, ER, PR and HER2/neu) was attempted. Results were subjected to statistical analysis. They were considered to be significant when the p-value was less than 0.05. RESULTS: The positivity for MUC1, MUC2 and MUC5AC in BC was 58%, 8% and 6% and for ER, PR and HER2 was 48%, 36% and 64% respectively. There was a significant correlation between MUC1 expression and ER and PR positivity. There was a significant correlation between MUC2 expression and ER positivity. No significant association was observed between MUC2 and PR expression, MUC5AC expression and ER and PR positivity. There was statistically significant correlation between negative MUC2 and MUC5AC expression and histopathological grade. It was noted that MUC2 and MUC5AC negative tumours were associated with higher tumour stage though not statistically significant. It was noted that MUC5AC negative tumours showed higher frequency of lymphovascular invasion though not statistically significant. CONCLUSION: Our experience with the present study highlights the role of mucins in the development and progression of BC.

Establishment of a novel cell line from intraductal papillary neoplasm of the bile duct.

BACKGROUND: Intraductal papillary neoplasm of the bile duct (IPNB) is a novel and increasingly-occurring disease. Its characteristics remain uncertain because of the lack of an in vitro culture system. We established and characterized a novel cell line from a human IPNB. MATERIALS AND METHODS: We obtained tumor tissue from a surgical specimen from a patient with IPNB. Cells were primary co-cultured with mouse stromal cells in serum-free medium. Tumor characteristics were compared among the primary IPNB, established cell line, and xenograft. RESULTS: We successfully established an IPNB cell line. We temporary termed this cell line Kobe Bile Duct Cancer (KBDC)-11. Xenograft formed a tumor which had ductal structures and mucus production as the primary tumor did. Overexpression of p53, MUC staining pattern, and CD133 expression were common among the primary IPNB, KBDC-11, and the xenograft. CONCLUSION: This novel cell line established from IPNB exhibited the same features as IPNB and might contribute to studies of IPNB and its process of malignant transformation.

Mucin profile expression in gastric adenocarcinoma.

BACKGROUND: The process of neoplastic transformation in the stomach has been reported to be associated with decreased expression of normal mucins of the gastric mucosa and denovo expression of mucins that are normally expressed in other organs. This association may be used as a means to bring new insights into biologic behavior and genetic pathways in the development of gastric cancer. There are controversial reports about differences in the genetic pathway and behavior of gastric cancer in younger patients in comparison with older ones. This study aims to determine if there is any difference in mucin profiles between different age groups with gastric cancer. METHODS: Over a five-year (2003-2008) period, 43 cases of gastric cancer (≤50) years were referred to our center. Of these, 40 had adequate tissue for additional study, whereas three cases lacked a sufficient amount of tumor tissue for immunohistochemistry (IHC) analysis. A group of 40 gastric cancer patients above the age of 50 years were gender-matched with the first group. Expressions of MUC-1, MUC-2, MUC-5AC, and MUC-6 were evaluated by IHC for the total 80 gastric cancer cases. RESULTS: The expressions of the mucins did not show a significant difference between the two age groups. CONCLUSION: Gastric cancer in both young and old age adults was not significantly different in terms of mucin profiles. Our results have shown that younger age is not predictive of gastric cancer phenotype, which can be an indicator of the lack of difference in the genetic pathways and molecular alterations in these two age groups.

Upregulation of Intestinal Mucin Expression by the Probiotic Bacterium E. coli Nissle 1917.

The probiotic E. coli Nissle 1917 (EcN) has been reported to have various health benefits; however, very little is known about their underlying mechanisms. In this regard, the present study aimed to elucidate the effect of the bacterium on mucin production by intestinal epithelial cells. Incubation of HT-29 cells with EcN lead to a contact time-dependent rise in mRNA levels of the MUC2, MUC3, MUC5AC, and MUC5A. The expression was markedly higher with MUC5AC gene. In most cases, MUC genes expression was more pronounced in polarized cells compared to non-polarized ones. In contrast to MUC3, the basal stimulation of polarized cells brought about markedly higher levels of other tested mucins. Similar but milder results were observed when living EcN was replaced by inactivated bacteria. With exception of MUC3, the conditioned media showed no significant effect on the mRNA level of the tested mucins. The above-mentioned mRNA results were confirmed on protein level using enzyme-linked lectin assay (ELLA) and enzyme-linked immunosorbant assay (ELISA). In contrast to other treatments, basal stimulation of polarized cells showed a growth phase-dependent MUC induction with more prominent effect by stationary-phase bacteria. In contrast to MUC 2 and MUC3, the induction of MUC5AC and MUC5B showed a bacterial count-dependent pattern. In conclusion, EcN was found to stimulate MUC gene expression in HT-29 intestinal cells. This stimulation was more distinct with polarized cells. Such observation may partially interpret some health benefits of the probiotic bacterium including antagonizing pathogen adhesion and protection of the intestinal mucosa.

Intraductal neoplasm of the intrahepatic bile duct: clinicopathological study of 24 cases.

AIM: To investigate the clinicopathological features of intraductal neoplasm of the intrahepatic bile duct (INihB). METHODS: Clinicopathological features of 24 cases of INihB, which were previously diagnosed as biliary papillomatosis or intraductal growth of intrahepatic biliary neoplasm, were reviewed. Mucin immunohistochemistry was performed for mucin (MUC)1, MUC2, MUC5AC and MUC6. Ki-67, P53 and ß-catenin immunoreactivity were also examined. We categorized each tumor as adenoma (low grade), borderline (intermediate grade), and malignant (carcinoma in situ, high grade including tumors with microinvasion). RESULTS: Among 24 cases of INihB, we identified 24 tumors. Twenty of 24 tumors (83%) were composed of a papillary structure; the same feature observed in intraductal papillary neoplasm of the bile duct (IPNB). In contrast, the remaining four tumors (17%) showed both tubular and papillary structures. In three of the four tumors (75%), macroscopic mucin secretion was limited but microscopic intracellular mucin was evident. Histologically, 16 tumors (67%) were malignant, three (12%) were borderline, and five (21%) were adenoma. Microinvasion was found in four cases (17%). Immunohistochemical analysis revealed that MUC1 was not expressed in the borderline/adenoma group but was expressed only in malignant lesions (P = 0.0095). Ki-67 labeling index (LI) was significantly higher in the malignant group than in the borderline/adenoma group (22.2 ± 15.5 vs 7.5 ± 6.3, P < 0.01). In the 16 malignant cases, expression of MUC5AC showed borderline significant association with high Ki-67 LI (P = 0.0622). Nuclear expression of ß-catenin was observed in two (8%) of the 24 tumors, and these two tumors also showed MUC1 expression. P53 was negative in all tumors. CONCLUSION: Some cases of INihB have a tubular structure, and are subcategorized as IPNB with tubular structure. MUC1 expression in INihB correlates positively with degree of malignancy.

Expresión de mucinas: MUC1, MUC2, MUC5AC, MUC6, HGM y CD10 en carcinomas gástricos y lesiones pre-neoplasicas adyacentes / MUCIN EXPRESSION: MUC1, MUC2, MUC5AC, MUC6, HGM AND CD10 IN GASTRIC CARCINOMAS AND ADJACENT PRE-NEOPLASTIC LESIONS

Introducción: Las mucinas son glicoproteínas y desempeñan funciones biológicas. Diversas anormalidades genéticas y epgenéticas han sido descritas en el cáncer gástrico. El objetivo de la investigación fue observar la inmuno-expresión de mucinas en cánceres difusos e intestinales y las lesiones pre-neoplásicas limítrofes. Métodos: Se evaluaron 18 cánceres difusos (56.3%) y 14 intestinales (43.7%) y las lesiones precursoras adyacentes al cáncer gástrico. Se realizó inmunohistoquímica para los siguientes marcadores: MUC-1, MUC-2, MUC5-AC, MUC-6, HGM y CD10. Resultados: La inmune-expresiones fue: MUC-1 (54.5%), de los cánceres intestinales y (45.5%) de los cánceres difusos. MUC-2 (50%) de los cánceres difusos y (50%) de los cánceres del tipo intestinal. MUC-5AC (39.3%) del tipo difuso (60.7%) del tipo intestinal. HGM (37.5%) del tipo intestinal y (62.5%) del tipo difuso. MUC-6 (57.9%) del tipo difuso (42.1%) los del tipo intestinal. CD10 (55.6%) del tipo intestinal, y (44.4%) en los difusos.En las lesiones pre- cursoras adyacentes al cáncer gástrico la MUC-1 se inmunoexpresa en metaplasia intestinal (9.7%). MUC-2 (83.9%) en metaplasia intestinal. MUC5-AC (90,6%) en foveolas MUC-6 (100%) positiva en glándulas. CD10 (54.8%) positiva en metaplasia intestinal.HGM (75%) en foveolas y el (64.5%) en metaplasia intestinal. MUC-6 (100%) en glándulas profundas y (64,5%) en metaplasia intestinal. Las dis- plasias expresaron MUC-2 y MUC-5AC, en el 80% y 100% respectivamente. Conclusiones: La inmunotipificación del cáncer gástrico permitirá una clasificación más exacta de los tumores asi como la identificación de posibles dianas terapéuticas y su relación con factores genético y epigeneticos.

Introduction: Mucins are glycoproteins and has diverse biological roles. Epi-genetic abnormalities have been described in gastric cancer. The aim of the research was evaluate immunoexpression of mucins in diffuse and intestinal cancers and pre-neoplastic lesions. Methods: We evaluated 18 diffuse cancers (56.3%) and 14 intestinal (43.7%) and adjacent precursor lesions to gastric cancer immunohistochemical markers used was MUC-1, MUC-2-AC MUC5, MUC-6, and CD10 HGM. Results: The immuno-expression was: MUC-1 (54.5%), and intestinal cancers (45.5%) diffuse cancers. MUC-2 (50%) and diffuse cancers (50%) of cancers of the intestinal type. MUC-5AC (39.3%) diffuse type (60.7%) of intestinal type.HGM (37.5%) intestinal type (62.5%) in diffuse type. MUC-6 (57.9%) diffuse type (42.1%) in the intestinal type. CD10 (55.6%) in intestinal type, and (44.4%) in the diffuse type In adjacent precursor lesions to gastric cancer we observed MUC-1 in intestinal metaplasia (9.7%). MUC-2 (83.9%) in intestinal metaplasia. MUC5-AC (90.6%) in foveolasMUC-6 (100%) positive glands. CD10 (54.8%) positive in intestinal.HGM metaplasia(75%) and in foveolae (64.5%) in metaplasia intestinal. MUC-6 (100%) deep glands (64.5%) in metaplasia.intestinal. Dys- plasias expressed MUC-2 and MUC-5AC, 80% and 100% respectively. Conclusions: The gastric cancer immunotyping allow more accurate classification of tumors and the identification of potential therapeutic targets and its relationship with genetic and epigenetic factors.

Precursor lesions of pancreatic cancer.

This review article describes morphological aspects, gene abnormalities, and mucin expression profiles in precursor lesions such as pancreatic intraepithelial neoplasia (PanIN), intraductal papillary mucinous neoplasm (IPMN), and mucinous cystic neoplasm (MCN) of the pancreas, as well as their relation to pancreatic ductal adenocarcinoma (PDAC). The gene abnormalities in precursors of PDAC are summarized as follows: (1) KRAS mutation and p16/CDKN2A inactivation are early events whose frequencies increase with the dysplasia grade in both PanIN and IPMN; (2) TP53 mutation and SMAD4/DPC4 inactivation are late events observed in PanIN3 or carcinomatous change of IPMN in both PanIN and IPMN, although the frequency of the TP53 mutation is lower in IPMN than in PDAC; and (3) also in MCN, KRAS mutation is an early event whose frequency increases with the dysplasia grade, whereas TP53 mutation and SMAD4/DPC4 inactivation are evident only in the carcinoma. The mucin expression profiles in precursors of PDAC are summarized as follows: (1) MUC1 expression increases with the PanIN grade, and is high in PDAC; (2) the expression pattern of MUC2 differs markedly between the major subtypes of IPMN with different malignancy potentials (i.e., IPMN-intestinal type with MUC2+ expression and IPMN-gastric type with MUC2- expression); (3) MUC2 is not expressed in any grade of PanINs, which is useful for differentiating PanIN from intestinal-type IPMN; (4) de novo expression of MUC4, which appears to increase with the dysplasia grade; and (5) high de novo expression of MUC5AC in all grades of PanINs, all types of IPMN, MCN, and PDAC.