Identification of subgroup-specific miRNA patterns by epigenetic profiling of sporadic and Lynch syndrome-associated colorectal and endometrial carcinoma

Abstract Background Altered expression of microRNAs (miRNAs) commonly accompanies colorectal (CRC) and endometrial carcinoma (EC) development, but the underlying mechanisms and clinicopathological correlations remain to be clarified. We focused on epigenetic mechanisms and aimed to explore if DNA methylation patterns in tumors depend on DNA mismatch repair (MMR) status, sporadic vs. Lynch-associated disease, and geographic origin (Finland vs. Australia). Treatment of cancer cell lines with demethylating agents revealed 109 significantly upregulated miRNAs. Seven met our stringent criteria for possible methylation-sensitive miRNAs and were used to screen patient specimens (205 CRCs and 36 ECs) by methylation-specific multiplex ligation-dependent probe amplification. Results Three miRNAs (129-2, 345, and 132) with low methylation levels in normal tissue and frequent hypermethylation in tumors were of particular interest. Hypermethylation of miR-345 and miR-132 associated with MMR deficiency in CRC regardless of geographic origin, and hypermethylation of miR-132 distinguished sporadic MMR-deficient CRC from Lynch-CRC. Finally, hypermethylation of miRNAs stratified 49 endometrial hyperplasias into low-methylator (simple hyperplasia) and high-methylator groups (complex hyperplasia with or without atypia) and suggested that miR-129-2 methylation in particular could serve as a marker of progression in early endometrial tumorigenesis. Conclusions Our study identifies miR-345 and miR-132 as novel differentially methylated miRNAs in CRC, thereby facilitating sub-classification of CRC and links miR-129-2 methylation to early endometrial tumorigenesis

Distinct microbiota dysbiosis in patients with non-erosive reflux disease and esophageal adenocarcinoma

Non-erosive reflux disease (NERD) and esophageal adenocarcinoma (EAC) are often regarded as bookends in the gastroesophageal reflux disease spectrum. However, there is limited clinical evidence to support this disease paradigm while the underlying mechanisms of disease progression remain unclear. In this study, we used 16S rRNA sequencing and mass-spectrometer-based proteomics to characterize the esophageal microbiota and host mucosa proteome, respectively. A total of 70 participants from four patient groups (NERD, reflux esophagitis, Barrett’s esophagus, and EAC) and a control group were analyzed. Our results showed a unique NERD microbiota composition, distinct to control and other groups. We speculate that an increase in sulfate-reducing Proteobacteria and Bacteroidetes along with hydrogen producer Dorea are associated with a mechanistic role in visceral hypersensitivity. We also observed a distinct EAC microbiota consisting of a high abundance of lactic acid-producing bacteria (Staphylococcus, Lactobacillus, Bifidobacterium, and Streptococcus), which may contribute towards carcinogenesis through dysregulated lactate metabolism. This study suggests the close relationship between esophageal mucosal microbiota and the appearance of pathologies of this organ

Long-term management of patients taking proton pump inhibitors

Proton pump inhibitors have changed the management of acid-related upper gastrointestinal disorders. Other effective strategies for reducing upper gastrointestinal morbidity include lifestyle modification, Helicobacter pylori eradication for patients with present or past peptic ulcer disease and infection, and less potent therapy for mild dyspepsia and gastro-oesophageal reflux. Proton pump inhibitors have a definite role in the prevention of recurrence of oesophageal strictures. They can also be used to prevent the ulcerative complications of non-steroidal anti-inflammatory drugs in patients at high risk. In Barrett's oesophagus the efficacy of proton pump inhibitors in preventing disease progression and the development of adenocarcinoma is unclear

The “Mutated in Colorectal Cancer” Protein Is a Novel Target of the UV-Induced DNA Damage Checkpoint

MCC is a potential tumor suppressor gene, which is silenced by promoter hypermethylation in a subset of colorectal cancers. However, its functions have remained poorly understood. In the present study, we describe a novel function of MCC in the DNA damage response. Several novel phosphorylation sites were identified by mass spectrometry, including 2 highly conserved ATM/ATR consensus sites at serine 118 and serine 120. In addition, exposure to ultraviolet radiation (UV), but not phleomycin, caused PI3K-dependent phosphorylation of MCC and its nuclear localization. Re-expression of MCC in HCT15 colorectal cancer cells led to a G2/M arrest, and MCC knockdown impaired the induction of a G2/M arrest following UV radiation. Finally, mutation of S118/120 to alanine did not affect MCC nuclear shuttling following UV but did impair MCC G2/M checkpoint activity. Thus, these results suggest that MCC is a novel target of the DNA damage checkpoint and that MCC is required for the complete cell cycle arrest in the G2/M phase in response to UV

Loss of special AT-rich binding protein 1 expression is a marker of poor survival in lung cancer

Introduction: Lung cancer is the leading cause of cancer-related mortality and requires more effective molecular markers of prognosis and therapeutic responsiveness. Special AT-rich binding protein 1 (SATB1) is a global genome organizer that recruits chromatin remodeling proteins to epigenetically regulate hundreds of genes in a tissue-specific manner. Initial studies suggest that SATB1 overexpression is a predictor of poor prognosis in breast cancer, but the prognostic significance of SATB1 expression has not been evaluated in lung cancer. Methods: A cohort of 257 lung cancers was evaluated by immunohistochemistry. Epigenetic silencing of SATB1 was examined in cell lines by 5-Aza 2-deoxycytidine and trichostatin A treatment, and chromatin immunoprecipitation. Results: Significant loss of SATB1 expression was found in squamous preinvasive lesions (p < 0.04) and in non-small cell lung cancers (p < 0.001) compared with matched normal bronchial epithelium. Loss of SATB1 independently predicted poor cancer-specific survival in squamous cell carcinomas (SCCs; hazard ratio: 2.06, 95% confidence interval: 1.2-3.7, p = 0.016). Treatment of lung cancer cell lines with the histone deacetylase inhibitor trichostatin A resulted in up-regulation of SATB1. SATB1 was associated with a decrease in the active chromatin mark acetylated histone H3K9 and an increase in the repressive polycomb mark trimethylated H3K27 in a SCC cell line relative to a normal bronchial epithelial cell line. Conclusions: This is the first study showing that SATB1 expression is lost in early preinvasive squamous lesions and that loss of SATB1 is associated with poor prognosis in lung SCC. We hypothesize that the SATB1 gene is epigenetically silenced through histone modifications

'MCC' protein interacts with E-cadherin and β-catenin strengthening cell-cell adhesion of HCT116 colon cancer cells

E-cadherin and β-catenin are key proteins that are essential in the formation of the epithelial cell layer in the colon but their regulatory pathways that are disrupted in cancer metastasis are not completely understood. Mutated in colorectal cancer (MCC) is a tumour suppressor gene that is silenced by promoter methylation in colorectal cancer and particularly in patients with increased lymph node metastasis. Here, we show that MCC methylation is found in 45% of colon and 24% of rectal cancers and is associated with proximal colon, poorly differentiated, circumferential and mucinous tumours as well as increasing T stage and larger tumour size. Knockdown of MCC in HCT116 colon cancer cells caused a reduction in E-cadherin protein level, which is a hallmark of epithelial–mesenchymal transition in cancer, and consequently diminished the E-cadherin/β-catenin complex. MCC knockdown disrupted cell–cell adhesive strength and integrity in the dispase and transepithelial electrical resistance assays, enhanced hepatocyte growth factor-induced cell scatter and increased tumour cell invasiveness in an organotypic assay. The Src/Abl inhibitor dasatinib, a candidate anti-invasive drug, abrogated the invasive properties induced by MCC deficiency. Mechanistically, we establish that MCC interacts with the E-cadherin/β-catenin complex. These data provide a significant advance in the current understanding of cell–cell adhesion in colon cancer cells

In vivo confocal endomicroscopy in the diagnosis and evaluation of celiac disease

Copyright © 2008 AGA Institute Published by Elsevier Inc.Background & Aims Accurate histopathology of endoscopic duodenal biopsy specimens is critical in the diagnosis of celiac disease (CD) but sampling error and poor quality specimens may generate a false-negative result. Confocal endomicroscopy (CEM) is a novel technology allowing real-time in vivo microscopy of the mucosa that may diagnose CD and evaluate its severity and response to treatment more accurately than histopathology. Methods Subjects with CD and controls prospectively underwent CEM. Features of villous atrophy and crypt hypertrophy were defined. A CEM score measuring CD severity was devised and validated against the diagnosis of CD and blinded histopathology. Receiver operator characteristics, sensitivity to change after treatment, and reliability of findings were assessed. Results From 31 patients (6 untreated CD, 11 treated CD, and 14 controls), 7019 CEM images paired with 326 biopsy specimens were obtained. The accuracy of CEM in diagnosing CD was excellent (receiver operator characteristics area under the curve, 0.946; sensitivity, 94%, specificity, 92%) and correlated well with the Marsh grading (R-squared, 0.756). CEM differentiated CD from controls (P &lt; .0001) and was sensitive to change after treatment with gluten-free diet (1787 optical biopsies; P = .012). The intraclass correlation of reliability was high (0.759–0.916). Of the 17 cases with diagnosed CD, 16 (94%) were diagnosed correctly using CEM but only 13 (76%) had detectable histopathology changes. The procedure was safe and well-tolerated. Conclusions CEM effectively diagnoses and evaluates CD severity in vivo. This promising technique has the potential to improve endoscopy efficiency.Rupert W.L. Leong, Nam Q. Nguyen, Christopher G. Meredith, Sam Al–Sohaily, Darko Kukic, Peter M. Delaney, Elise R. Murr, Jim Yong, Neil D. Merrett and Andrew V. Bianki