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Zhe Wang Department of Gastroenterology, Tongji Hospital, Tongji University School of Medicine, Shanghai, China
Division of Gastroenterology, Department of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

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Ke Ma Division of Gastroenterology, Department of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

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Steffie Pitts Cellular and Molecular Medicine Graduate Program, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

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Yulan Cheng Division of Gastroenterology, Department of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

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Xi Liu Department of Pathology, The First Affiliated Hospital of Xi’ an Jiaotong University, Xi’ an, Shaanxi, China

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Xiquan Ke Department of Gastroenterology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, China

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Samuel Kovaka Department of Computer Science, Whiting School of Engineering, Johns Hopkins University, Baltimore, Maryland, USA

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Hassan Ashktorab Cancer Center, Howard University School of Medicine, Washington, District of Columbia, USA

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Duane T Smoot Department of Medicine, Meharry Medical College, Nashville, Tennessee, USA

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Michael Schatz Department of Computer Science, Whiting School of Engineering, Johns Hopkins University, Baltimore, Maryland, USA

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Zhirong Wang Department of Gastroenterology, Tongji Hospital, Tongji University School of Medicine, Shanghai, China

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Stephen J Meltzer Division of Gastroenterology, Department of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

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Circular RNAs (circRNAs) are a new class of RNA involved in multiple human malignancies. However, limited information exists regarding the involvement of circRNAs in gastric carcinoma (GC). Therefore, we sought to identify novel circRNAs, their functions and mechanisms in gastric carcinogenesis. We analyzed next-generation RNA sequencing data from GC tissues and cell lines, identifying 75,201 candidate circRNAs. Among these, we focused on one novel circRNA, circNF1 , which was upregulated in GC tissues and cell lines. Loss- and gain-of-function studies demonstrated that circNF1 significantly promotes cell proliferation. Furthermore, luciferase reporter assays showed that circNF1 binds to miR-16, thereby derepressing its downstream target mRNAs, MAP7 and AKT3. Targeted silencing or overexpression of circNF1 had no effect on levels of its linear RNA counterpart, NF1. Taken together, these results suggest that circNF1 acts as a novel oncogenic circRNA in GC by functioning as a miR-16 sponge.

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Rachana Agarwal Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Zhe Jin Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Jian Yang Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Yuriko Mori Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Jee Hoon Song Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Sahil Kumar Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Masato Sato Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Yulan Cheng Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Alexandru V Olaru Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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John M Abraham Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Amit Verma Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Stephen J Meltzer Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine
Division of Gastroenterology, Department of Oncology, Department of Pathology, All India Institute of Medical Sciences, Tohoku University School of Medicine, Department of Medicine, Department of Medicine

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Yuriko Mori Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Alexandru V Olaru Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Yulan Cheng Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Rachana Agarwal Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Jian Yang Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Delgermaa Luvsanjav Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Wayne Yu Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Florin M Selaru Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Susan Hutfless Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Mark Lazarev Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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John H Kwon Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Steven R Brant Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Michael R Marohn Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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David F Hutcheon Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Mark D Duncan Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Ajay Goel Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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Stephen J Meltzer Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine
Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine
Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Division of Gastroenterology, Section of Gastroenterology, Department of Surgery, Division of Gastroenterology, Department of Medicine

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DNA hypermethylation is a common epigenetic abnormality in colorectal cancers (CRCs) and a promising class of CRC screening biomarkers. We conducted a genome-wide search for novel neoplasia-specific hypermethylation events in the colon. We applied methylation microarray analysis to identify loci hypermethylated in 17 primary CRCs relative to eight non-neoplastic colonic mucosae (NCs) from neoplasia-free subjects. These CRC-associated hypermethylation events were then individually evaluated for their ability to discriminate neoplastic from non-neoplastic cases, based on real-time quantitative methylation-specific PCR (qMSP) assays in 113 colonic tissues: 51 CRCs, nine adenomas, 19 NCs from CRC patients (CRC–NCs), and 34 NCs from neoplasia-free subjects (control NCs). A strict microarray data filtering identified 169 candidate CRC-associated hypermethylation events. Fourteen of these 169 loci were evaluated using qMSP assays. Ten of these 14 methylation events significantly distinguished CRCs from age-matched control NCs (P<0.05 by receiver operator characteristic curve analysis); methylation of visual system homeobox 2 (VSX2) achieved the highest discriminative accuracy (83.3% sensitivity and 92.3% specificity, P<1×10−6), followed by BEN domain containing 4 (BEND4), neuronal pentraxin I (NPTX1), ALX homeobox 3 (ALX3), miR-34b, glucagon-like peptide 1 receptor (GLP1R), BTG4, homer homolog 2 (HOMER2), zinc finger protein 583 (ZNF583), and gap junction protein, gamma 1 (GJC1). Adenomas were significantly discriminated from control NCs by hypermethylation of VSX2, BEND4, NPTX1, miR-34b, GLP1R, and HOMER2 (P<0.05). CRC–NCs were significantly distinguished from control NCs by methylation of ALX3 (P<1×10−4). In conclusion, systematic methylome-wide analysis has identified ten novel methylation events in neoplastic and non-neoplastic colonic mucosae from CRC patients. These potential biomarkers significantly discriminate CRC patients from controls. Thus, they merit further evaluation in stool- and circulating DNA-based CRC detection studies.

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